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http://www.ncbi.nlm.nih.gov/pubmed/10391249
1. Nature. 1999 Jun 24;399(6738):806-9. doi: 10.1038/21690. The tyrosine kinase c-Abl regulates p73 in apoptotic response to cisplatin-induced DNA damage. Gong JG(1), Costanzo A, Yang HQ, Melino G, Kaelin WG Jr, Levrero M, Wang JY. Author information: (1)Department of Biology, University of California, San Diego, La Jolla 92093-0322, USA. Comment in Nature. 1999 Jun 24;399(6738):734-5, 737. doi: 10.1038/21539. Cancer chemotherapeutic agents such as cisplatin exert their cytotoxic effect by inducing DNA damage and activating programmed cell death (apoptosis). The tumour-suppressor protein p53 is an important activator of apoptosis. Although p53-deficient cancer cells are less responsive to chemotherapy, their resistance is not complete, which suggests that other apoptotic pathways may exist. A p53-related gene, p73, which encodes several proteins as a result of alternative splicing, can also induce apoptosis. Here we show that the amount of p73 protein in the cell is increased by cisplatin. This induction of p73 is not seen in cells unable to carry out mismatch repair and in which the nuclear enzyme c-Abl tyrosine kinase is not activated by cisplatin. The half-life of p73 is prolonged by cisplatin and by co-expression with c-Abl tyrosine kinase; the apoptosis-inducing function of p73 is also enhanced by the c-Abl kinase. Mouse embryo fibroblasts deficient in mismatch repair or in c-Abl do not upregulate p73 and are more resistant to killing by cisplatin. Our results indicate that c-Abl and p73 are components of a mismatch-repair-dependent apoptosis pathway which contributes to cisplatin-induced cytotoxicity. DOI: 10.1038/21690 PMID: 10391249 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/24563699
1. Commun Integr Biol. 2013 Nov 1;6(6):e25660. doi: 10.4161/cib.25660. Epub 2013 Jul 23. Targeting secretion to the apical surface by mDia1-built actin tracks. Geron E(1), Schejter ED(1), Shilo BZ(1). Author information: (1)Department of Molecular Genetics; Weizmann Institute of Science; Rehovot, Israel. The apical surface of secretory tubular epithelia is a dynamic cellular domain where massive membrane turnover takes place during exocytosis and its subsequent compensatory endocytosis. This extensive membrane flow poses a difficulty in targeting secretory vesicles efficiently to a narrow apical domain. We have studied how actin filaments mediate the secretory process in the murine exocrine pancreas, which produces and secretes digestive enzymes that are deposited into the intestine. We show that cargo-filled secretory vesicles move over bundles of linear actin cables from their storage areas to the apical membrane of pancreatic acinar cells. mDia1, a linear actin nucleator of the Formin family, was identified as the generator of these structures. The active form of mDia1 localizes to the apical surface, and the microfilament bundles it forms emanate from the apical surface and extend into the cytoplasm, generating polarized secretion tracks. These bundles ensure orderly progression of exocytosis, since the apical targeting of pancreatic vesicles is compromised in their absence, and vesicles fuse with each other to generate compound, membrane-associated secretory structures. DOI: 10.4161/cib.25660 PMCID: PMC3917947 PMID: 24563699
http://www.ncbi.nlm.nih.gov/pubmed/19160018
1. Cancer Metastasis Rev. 2009 Jun;28(1-2):65-76. doi: 10.1007/s10555-008-9170-7. Rho signaling, ROCK and mDia1, in transformation, metastasis and invasion. Narumiya S(1), Tanji M, Ishizaki T. Author information: (1)Department of Pharmacology, Kyoto University Faculty of Medicine, Kyoto, 606-8501, Japan. snaru@mfour.med.kyoto-u.ac.jp The Rho subgroup of the Rho GTPases consisting of RhoA, RhoB and RhoC induces a specific type of actin cytoskeleton and carry out a variety of functions in the cell. mDia and ROCK are downstream effectors of Rho mediating Rho action on the actin cytoskeleton; mDia produces actin filaments by nucleation and polymerization and ROCK activate myosin to cross-link them for induction of actomyosin bundles and contractility. mDia is potentially linked to Rac activation and membrane ruffle formation through c-Src-induced phosphorylation of focal adhesion proteins, and ROCK antagonizes this mDia action. Thus, cell morphogenesis, adhesion, and motility can be determined by the balance between mDia and ROCK activities. Though they are not oncogenes by themselves, overexpression of RhoA and RhoC are often found in clinical cancers, and RhoC has been repeatedly identified as a gene associated with metastasis. The Rho-ROCK pathway is implicated in Ras-mediated transformation, the amoeboid movement of tumor cells in the three-dimensional matrix, and transmigration of tumor cells through the mesothelial monolayer. On the other hand, the Rho-mDia1 pathway is implicated in Src-mediated remodeling of focal adhesions and migration of tumor cells. There is also an indication that the Rho pathway other than ROCK is involved in Src-mediated induction of podosome and regulation of matrix metalloproteases. Thus, Rho mediates various phenotypes of malignant transformation by Ras and Src through its effectors, ROCK and mDia. DOI: 10.1007/s10555-008-9170-7 PMID: 19160018 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/26109050
1. Genes Dev. 2015 Jun 15;29(12):1271-84. doi: 10.1101/gad.262816.115. A YAP/TAZ-induced feedback mechanism regulates Hippo pathway homeostasis. Moroishi T(1), Park HW(1), Qin B(2), Chen Q(3), Meng Z(1), Plouffe SW(1), Taniguchi K(4), Yu FX(5), Karin M(4), Pan D(3), Guan KL(1). Author information: (1)Department of Pharmacology, Moores Cancer Center, University of California at San Diego, La Jolla, California 92093, USA; (2)Department of Pharmacology, Moores Cancer Center, University of California at San Diego, La Jolla, California 92093, USA; Department of Laboratory Medicine, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai 200003, China; (3)Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA; (4)Department of Pharmacology, University of California at San Diego, La Jolla, California 92093, USA; Department of Pathology, University of California at San Diego, La Jolla, California 92093, USA; (5)Children's Hospital, Fudan University, Shanghai 200032, China; Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China. YAP (Yes-associated protein) and TAZ (transcriptional coactivator with PDZ-binding motif) are major downstream effectors of the Hippo pathway that influences tissue homeostasis, organ size, and cancer development. Aberrant hyperactivation of YAP/TAZ causes tissue overgrowth and tumorigenesis, whereas their inactivation impairs tissue development and regeneration. Dynamic and precise control of YAP/TAZ activity is thus important to ensure proper physiological regulation and homeostasis of the cells. Here, we show that YAP/TAZ activation results in activation of their negative regulators, LATS1/2 (large tumor suppressor 1/2) kinases, to constitute a negative feedback loop of the Hippo pathway in both cultured cells and mouse tissues. YAP/TAZ in complex with the transcription factor TEAD (TEA domain family member) directly induce LATS2 expression. Furthermore, YAP/TAZ also stimulate the kinase activity of LATS1/2 through inducing NF2 (neurofibromin 2). This feedback regulation is responsible for the transient activation of YAP upon lysophosphatidic acid (LPA) stimulation and the inhibition of YAP-induced cell migration. Thus, this LATS-mediated feedback loop provides an efficient mechanism to establish the robustness and homeostasis of YAP/TAZ regulation. © 2015 Moroishi et al.; Published by Cold Spring Harbor Laboratory Press. DOI: 10.1101/gad.262816.115 PMCID: PMC4495398 PMID: 26109050 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/25109332
1. Oncogene. 2015 Jun 11;34(24):3095-106. doi: 10.1038/onc.2014.251. Epub 2014 Aug 11. Geranylgeranylation signals to the Hippo pathway for breast cancer cell proliferation and migration. Mi W(1), Lin Q(2), Childress C(1), Sudol M(3), Robishaw J(1), Berlot CH(1), Shabahang M(4), Yang W(1). Author information: (1)Weis Center for Research, Danville, PA, USA. (2)1] Weis Center for Research, Danville, PA, USA [2] School of Medical Sciences and Laboratory Medicine, Jiangsu University, Zhenjiang, China. (3)1] Weis Center for Research, Danville, PA, USA [2] Department of Medicine, Mount Sinai Medical School, New York, NY, USA. (4)Department of General Surgery, Geisinger Clinic, Danville, PA, USA. Protein geranylgeranylation (GGylation) is an important biochemical process for many cellular signaling molecules. Previous studies have shown that GGylation is essential for cell survival in many types of cancer. However, the molecular mechanism mediating the cell survival effect remains elusive. In this report, we show that the Hippo pathway mediates GGylation-dependent cell proliferation and migration in breast cancer cells. Blockade of GGylation enhanced phosphorylation of Mst1/2 and Lats1, and inhibited YAP and TAZ activity and the Hippo-YAP/TAZ pathway-dependent transcription. The effect of GGylation blockade on inhibition of breast cancer cell proliferation and migration is dependent on the Hippo-YAP/TAZ signaling, in which YAP appears to regulate cell proliferation and TAZ to regulate cell migration. Furthermore, GGylation-dependent cell proliferation is correlated with the activity of YAP/TAZ in breast cancer cells. Finally, Gγ and RhoA are the GGylated proteins that may transduce GGylation signals to the Hippo-YAP/TAZ pathway. Taken together, our studies have demonstrated that the Hippo-YAP/TAZ pathway is essential for GGylation-dependent cancer cell proliferation and migration. DOI: 10.1038/onc.2014.251 PMID: 25109332 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/16818632
1. Cancer Res. 2006 Jul 1;66(13):6598-605. doi: 10.1158/0008-5472.CAN-05-3115. MYC can induce DNA breaks in vivo and in vitro independent of reactive oxygen species. Ray S(1), Atkuri KR, Deb-Basu D, Adler AS, Chang HY, Herzenberg LA, Felsher DW. Author information: (1)Division of Oncology, Department of Medicine and Pathology, Stanford University School of Medicine, 269 Campus Drive, Stanford, CA 94305, USA. MYC overexpression is thought to initiate tumorigenesis by inducing cellular proliferation and growth and to be restrained from causing tumorigenesis by inducing cell cycle arrest, cellular senescence, and/or apoptosis. Here we show that MYC can induce DNA breaks both in vitro and in vivo independent of increased production of reactive oxygen species (ROS). We provide an insight into the specific circumstances under which MYC generates ROS in vitro and propose a possible mechanism. We found that MYC induces DNA double-strand breaks (DSBs) independent of ROS production in murine lymphocytes in vivo as well as in normal human foreskin fibroblasts (NHFs) in vitro in normal (10%) serum, as measured by gammaH2AX staining. However, NHFs cultured in vitro in low serum (0.05%) and/or ambient oxygen saturation resulted in ROS-associated oxidative damage and DNA single-strand breaks (SSBs), as measured by Ape-1 staining. In NHFs cultured in low versus normal serum, MYC induced increased expression of CYP2C9, a gene product well known to be associated with ROS production. Specific inhibition of CYP2C9 by small interfering RNA was shown to partially inhibit MYC-induced ROS production. Hence, MYC overexpression can induce ROS and SSBs under some conditions, but generally induces widespread DSBs in vivo and in vitro independent of ROS production. DOI: 10.1158/0008-5472.CAN-05-3115 PMID: 16818632 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/21930699
1. J Biol Chem. 2011 Nov 11;286(45):39236-46. doi: 10.1074/jbc.M111.253898. Epub 2011 Sep 19. Unique structural and nucleotide exchange features of the Rho1 GTPase of Entamoeba histolytica. Bosch DE(1), Wittchen ES, Qiu C, Burridge K, Siderovski DP. Author information: (1)Department of Pharmacology, University of North Carolina, Chapel Hill, North Carolina 27599-7365, USA. The single-celled human parasite Entamoeba histolytica possesses a dynamic actin cytoskeleton vital for its intestinal and systemic pathogenicity. The E. histolytica genome encodes several Rho family GTPases known to regulate cytoskeletal dynamics. EhRho1, the first family member identified, was reported to be insensitive to the Rho GTPase-specific Clostridium botulinum C3 exoenzyme, raising the possibility that it may be a misclassified Ras family member. Here, we report the crystal structures of EhRho1 in both active and inactive states. EhRho1 is activated by a conserved switch mechanism, but diverges from mammalian Rho GTPases in lacking a signature Rho insert helix. EhRho1 engages a homolog of mDia, EhFormin1, suggesting a role in mediating serum-stimulated actin reorganization and microtubule formation during mitosis. EhRho1, but not a constitutively active mutant, interacts with a newly identified EhRhoGDI in a prenylation-dependent manner. Furthermore, constitutively active EhRho1 induces actin stress fiber formation in mammalian fibroblasts, thereby identifying it as a functional Rho family GTPase. EhRho1 exhibits a fast rate of nucleotide exchange relative to mammalian Rho GTPases due to a distinctive switch one isoleucine residue reminiscent of the constitutively active F28L mutation in human Cdc42, which for the latter protein, is sufficient for cellular transformation. Nonconserved, nucleotide-interacting residues within EhRho1, revealed by the crystal structure models, were observed to contribute a moderating influence on fast spontaneous nucleotide exchange. Collectively, these observations indicate that EhRho1 is a bona fide member of the Rho GTPase family, albeit with unique structural and functional aspects compared with mammalian Rho GTPases. DOI: 10.1074/jbc.M111.253898 PMCID: PMC3234748 PMID: 21930699 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/14765113
1. EMBO J. 2004 Feb 25;23(4):760-71. doi: 10.1038/sj.emboj.7600095. Epub 2004 Feb 5. DIP (mDia interacting protein) is a key molecule regulating Rho and Rac in a Src-dependent manner. Meng W(1), Numazaki M, Takeuchi K, Uchibori Y, Ando-Akatsuka Y, Tominaga M, Tominaga T. Author information: (1)Department of Cellular and Molecular Physiology, Mie University School of Medicine, Tsu, Japan. Cell movement is driven by the coordinated regulation of cytoskeletal reorganization through Rho GTPases downstream of integrin and growth-factor receptor signaling. We have reported that mDia, a target protein of Rho, interacts with Src and DIP. Here we show that DIP binds to p190RhoGAP and Vav2, and that DIP is phosphorylated by Src and mediates the phosphorylation of p190RhoGAP and Vav2 upon EGF stimulation. When endogenous DIP was inhibited by expressing dominant-negative mutants of DIP or siRNA, phosphorylation of p190RhoGAP and Vav2 upon EGF stimulation was diminished, and EGF-induced actin organization, distribution of p190RhoGAP and Vav2, and cell movement were affected. Therefore, DIP seems to transfer the complex of the three proteins from cytosol to beneath the membrane, and the three proteins, in turn, can be phosphorylated by Src. DIP inactivated Rho and activated Rac following EGF stimulation in the membrane fraction. Thus, DIP acts as a regulatory molecule causing Src kinase-dependent feedback modulation of Rho GTPases downstream of Rho-mDia upon EGF stimulation, and plays an important role in cell motility. DOI: 10.1038/sj.emboj.7600095 PMCID: PMC381003 PMID: 14765113 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/8144827
1. J Am Geriatr Soc. 1994 Apr;42(4):413-9. doi: 10.1111/j.1532-5415.1994.tb07490.x. The effects of the presence of a third person on the physician-older patient medical interview. Greene MG(1), Majerovitz SD, Adelman RD, Rizzo C. Author information: (1)Department of Health and Nutrition Sciences, Brooklyn College, New York. OBJECTIVE: To compare communication in triadic (three-person) and dyadic (two-person) older patient medical interviews and to determine the influence of the presence of a third person on the physician-older patient relationship. DESIGN: Matched sample of dyadic and triadic audiotaped outpatient medical visits. Audiotapes were coded with the Multi-dimensional Interaction Analysis (MDIA) system. SETTING: Hospital-based medical primary care group practice in a major urban teaching institution. PARTICIPANTS: Patients 60 years and older who were making their first visit to study physicians. In a sample of 96 audiotaped initial medical visits, 15 encounters involved three persons. These 15 cases were matched with 15 dyadic interviews for gender and race of the patient and for gender and race of the physician. MAIN OUTCOME MEASURES: Content, interactional processes, and specific language and communication behaviors of older patients, physicians, and third persons in the medical encounter, as measured by the MDIA system. RESULTS: The specific content and the quality of interactional processes of physicians were not affected by the presence of a third person. However, older patients raised fewer topics in all content areas (medical, personal habits, psychosocial, and physician-patient relationship) in triads than in dyads. Overall, patients were less responsive (ie, the quality of their questioning, informing, and supportiveness was poorer) on patient-raised topics in triads than in dyads. Patients were rated as less assertive and expressive, and there was less joint decision-making and shared laughter in triads than in dyads. Patients were frequently excluded from conversations in visits in which a third person was present. CONCLUSIONS: The presence of a third person in the medical encounter changes the interactional dynamics of older patient medical interviews and may influence the development of a trusting and effective physician-older patient relationship. DOI: 10.1111/j.1532-5415.1994.tb07490.x PMID: 8144827 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/24242070
1. Cancer Res. 2013 Nov 15;73(22):6793-803. doi: 10.1158/0008-5472.CAN-13-1593. Small-molecule intramimics of formin autoinhibition: a new strategy to target the cytoskeletal remodeling machinery in cancer cells. Lash LL(1), Wallar BJ, Turner JD, Vroegop SM, Kilkuskie RE, Kitchen-Goosen SM, Xu HE, Alberts AS. Author information: (1)Authors' Affiliations: Laboratories of Cell Structure and Signal Integration and Structural Sciences, Van Andel Research Institute; Grand Valley State University, Grand Rapids; and Michigan High Throughput Screening Center, Kalamazoo, Michigan. Although the cancer cell cytoskeleton is a clinically validated target, few new strategies have emerged for selectively targeting cell division by modulating the cytoskeletal structure, particularly ways that could avoid the cardiotoxic and neurotoxic effects of current agents such as taxanes. We address this gap by describing a novel class of small-molecule agonists of the mammalian Diaphanous (mDia)-related formins, which act downstream of Rho GTPases to assemble actin filaments, and their organization with microfilaments to establish and maintain cell polarity during migration and asymmetric division. GTP-bound Rho activates mDia family members by disrupting the interaction between the DID and DAD autoregulatory domains, which releases the FH2 domain to modulate actin and microtubule dynamics. In screening for DID-DAD disruptors that activate mDia, we identified two molecules called intramimics (IMM-01 and -02) that were sufficient to trigger actin assembly and microtubule stabilization, serum response factor-mediated gene expression, cell-cycle arrest, and apoptosis. In vivo analysis of IMM-01 and -02 established their ability to slow tumor growth in a mouse xenograft model of colon cancer. Taken together, our work establishes the use of intramimics and mDia-related formins as a new general strategy for therapeutic targeting of the cytoskeletal remodeling machinery of cancer cells. ©2013 AACR DOI: 10.1158/0008-5472.CAN-13-1593 PMID: 24242070 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23740402
1. Commun Integr Biol. 2012 Nov 1;5(6):631-3. doi: 10.4161/cib.21686. LIN7-IRSp53: A novel pathway for filopodia and neurite formation? Ferrari I(1), Crespi A, Scita G, Pietrini G. Author information: (1)Department of BIOMETRA; Università degli Studi di Milano; CNR-Institute of Neuroscience; Milan, Italy. Filopodia are dynamic, actin-rich finger-like structures that protrude from the cell membrane and play important roles in cell migration and neurite initiation and outgrowth. The insulin receptor substrate protein of 53 kDa (IRSp53) and the mammalian Diaphanous members of the formin family of proteins (mDia) are two key players in the formation of filopodia and neurites. IRSp53 is an adaptor protein that acts at the membrane:actin interface, coupling membrane deformation with F-actin polymerization. mDia formin proteins, instead, can nucleate and polymerize linear actin filaments. Emerging genetic and biochemical evidence indicate that there are multiple and independent pathways leading to filopodium and neurite formation, but the precise molecular components of these pathways remain ill-defined. We recently identified the PDZ domain-containing protein LIN7 as a novel regulator of IRSp53. We further showed that the association between these two proteins is required to promote the formation of filopodia and neurites independently from mDia formin proteins, highlighting novel mechanisms of filopodia and neurite formation. DOI: 10.4161/cib.21686 PMCID: PMC3541334 PMID: 23740402
http://www.ncbi.nlm.nih.gov/pubmed/10436006
1. Mol Biol Cell. 1999 Aug;10(8):2481-91. doi: 10.1091/mbc.10.8.2481. Distinct actions and cooperative roles of ROCK and mDia in Rho small G protein-induced reorganization of the actin cytoskeleton in Madin-Darby canine kidney cells. Nakano K(1), Takaishi K, Kodama A, Mammoto A, Shiozaki H, Monden M, Takai Y. Author information: (1)Department of Molecular Biology and Biochemistry, Osaka University Medical School, Suita 565-0871, Japan. Rho, a member of the Rho small G protein family, regulates the formation of stress fibers and focal adhesions in various types of cultured cells. We investigated here the actions of ROCK and mDia, both of which have been identified to be putative downstream target molecules of Rho, in Madin-Darby canine kidney cells. The dominant active mutant of RhoA induced the formation of parallel stress fibers and focal adhesions, whereas the dominant active mutant of ROCK induced the formation of stellate stress fibers and focal adhesions, and the dominant active mutant of mDia induced the weak formation of parallel stress fibers without affecting the formation of focal adhesions. In the presence of C3 ADP-ribosyltransferase for Rho, the dominant active mutant of ROCK induced the formation of stellate stress fibers and focal adhesions, whereas the dominant active mutant of mDia induced only the diffuse localization of actin filaments. These results indicate that ROCK and mDia show distinct actions in reorganization of the actin cytoskeleton. The dominant negative mutant of either ROCK or mDia inhibited the formation of stress fibers and focal adhesions, indicating that both ROCK and mDia are necessary for the formation of stress fibers and focal adhesions. Moreover, inactivation and reactivation of both ROCK and mDia were necessary for the 12-O-tetradecanoylphorbol-13-acetate-induced disassembly and reassembly, respectively, of stress fibers and focal adhesions. The morphologies of stress fibers and focal adhesions in the cells expressing both the dominant active mutants of ROCK and mDia were not identical to those induced by the dominant active mutant of Rho. These results indicate that at least ROCK and mDia cooperatively act as downstream target molecules of Rho in the Rho-induced reorganization of the actin cytoskeleton. DOI: 10.1091/mbc.10.8.2481 PMCID: PMC25478 PMID: 10436006 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/15371418
1. J Biol Chem. 2004 Nov 26;279(48):50250-6. doi: 10.1074/jbc.M404429200. Epub 2004 Sep 14. Homo-oligomerization is essential for F-actin assembly by the formin family FH2 domain. Copeland JW(1), Copeland SJ, Treisman R. Author information: (1)Transcription Laboratory, Cancer Research UK, London Research Institute, Lincoln's Inn Fields Laboratories, 44 Lincoln's Inn Fields, London C2A 3PX, United Kingdom. jcopelan@uottawa.ca Formin proteins regulate the actin and microtubule cytoskeletons and also control the activity of the SRF transcription factor through depletion of the G-actin pool. Although the conserved formin homology 2 (FH2) domains of the mDia1 and Bni1 formins can nucleate actin polymerization in vitro, the activity of other FH2 domains and the relationship between actin polymerization and microtubule reorganization have been controversial. We show that, similar to the mDia1 FH2 domain, the FH2 domains of mDia2 and ld are sufficient for SRF activation in vivo. We demonstrate that an mDia1 mutant defective for microtubule rearrangement in vivo is also defective in SRF activation in vivo as well as actin polymerization in vitro and that the mDia2 FH2 domain promotes actin polymerization in vitro. Using co-immunoprecipitation, we show that mDia1 is oligomeric in its inactive autoinhibited state in vivo, that the active mDia1 and mDia2 FH2 domains form homo- but not hetero-oligomers in vivo, and that oligomerization is abolished by inactivating FH2 deletion and point mutations. Nevertheless, inactive mDia1 FH2 domain mutants retain the ability to interfere with cellular mDia activity. Our results show that self-oligomerization is essential for SRF activation in vivo and F-actin assembly in vitro and provide strong support for recent structural models of the FH2 domain. DOI: 10.1074/jbc.M404429200 PMID: 15371418 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/18362183
1. J Cell Biol. 2008 Mar 24;180(6):1245-60. doi: 10.1083/jcb.200708123. WASP family members and formin proteins coordinate regulation of cell protrusions in carcinoma cells. Sarmiento C(1), Wang W, Dovas A, Yamaguchi H, Sidani M, El-Sibai M, Desmarais V, Holman HA, Kitchen S, Backer JM, Alberts A, Condeelis J. Author information: (1)Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Yeshiva University, Bronx, NY 10461, USA. csarmien@aecom.yu.edu We examined the role of the actin nucleation promoters neural Wiskott-Aldrich syndrome protein (N-WASP) and WAVE2 in cell protrusion in response to epidermal growth factor (EGF), a key regulator in carcinoma cell invasion. We found that WAVE2 knockdown (KD) suppresses lamellipod formation and increases filopod formation, whereas N-WASP KD has no effect. However, simultaneous KD of both proteins results in the formation of large jagged protrusions with lamellar properties and increased filopod formation. This suggests that another actin nucleation activity is at work in carcinoma cells in response to EGF. A mammalian Diaphanous-related formin, mDia1, localizes at the jagged protrusions in double KD cells. Constitutively active mDia1 recapitulated the phenotype, whereas inhibition of mDia1 blocked the formation of these protrusions. Increased RhoA activity, which stimulates mDia1 nucleation, was observed in the N-WASP/WAVE2 KD cells and was shown to be required for the N-WASP/WAVE2 KD phenotype. These data show that coordinate regulation between the WASP family and mDia proteins controls the balance between lamellar and lamellipodial protrusion activity. DOI: 10.1083/jcb.200708123 PMCID: PMC2290849 PMID: 18362183 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/16361249
1. J Biol Chem. 2006 Feb 24;281(8):5084-93. doi: 10.1074/jbc.M509226200. Epub 2005 Dec 16. Biochemical characterization of the diaphanous autoregulatory interaction in the formin homology protein FHOD1. Schönichen A(1), Alexander M, Gasteier JE, Cuesta FE, Fackler OT, Geyer M. Author information: (1)Max Planck Institute for Molecular Physiology, Department of Physical Biochemistry, D-44227 Dortmund, Germany. Diaphanous related formins (DRFs) are cytoskeleton remodeling proteins that mediate specific upstream GTPase signals to regulate cellular processes such as cytokinesis, cell polarity, and organelle motility. Previous work on the Rho-interacting DRF mDia has established that the biological activity of DRFs is regulated by an autoinhibitory interaction of a C-terminal diaphanous autoregulatory domain (DAD) with the DRF N terminus. This autoinhibition is released upon competitive binding of an activated GTPase to the N terminus of the DRF. Analyzing autoregulation of the Rac1-interacting DRF FHOD1, we utilized in vitro binding studies to identify a 60-amino acid DAD at the protein C terminus that recognizes an N-terminal formin homology (FH) 3 domain. Importantly, the FH3 domain of FHOD1 does not overlap with the proposed Rac1-binding domain. The FHOD1 DAD was found to contain one functional hydrophobic autoregulatory motif, while a previously uncharacterized basic cluster that is conserved in all DRF family DADs also contributed to the FH3-DAD interaction. Simultaneous mutation of both motifs efficiently released autoinhibition of FHOD1 in NIH3T3 cells resulting in the formation of actin stress fibers and increased serum response element transcription. A second putative hydrophobic autoregulatory motif N-terminal of the DAD belongs to a unique FHOD subdomain of yet undefined function. NMR structural analysis and size exclusion chromatography experiments revealed that the FHOD1 DAD is intrinsically unstructured with a tendency for a helical conformation in the hydrophobic autoregulation motif. Together, these data suggest that in FHOD1, DAD acts as signal sequence for binding to the well folded and monomeric FH3 domain and imply an activation mechanism that differs from competitive binding of Rac1 and DAD to one interaction site. DOI: 10.1074/jbc.M509226200 PMID: 16361249 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/24068556
1. Nucleic Acids Res. 2014 Jan;42(1):128-36. doi: 10.1093/nar/gkt854. Epub 2013 Sep 24. Nucleosome positioning and kinetics near transcription-start-site barriers are controlled by interplay between active remodeling and DNA sequence. Parmar JJ(1), Marko JF, Padinhateeri R. Author information: (1)Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Mumbai 400076, India, Department of Molecular Biosciences, Northwestern University, Evanston, IL 60208, USA, Department of Physics and Astronomy, Northwestern University, Evanston, IL 60208, USA and Wadhwani Research Centre for Biosciences and Bioengineering, Indian Institute of Technology Bombay, Mumbai 400076, India. We investigate how DNA sequence, ATP-dependent chromatin remodeling and nucleosome-depleted 'barriers' co-operate to determine the kinetics of nucleosome organization, in a stochastic model of nucleosome positioning and dynamics. We find that 'statistical' positioning of nucleosomes against 'barriers', hypothesized to control chromatin structure near transcription start sites, requires active remodeling and therefore cannot be described using equilibrium statistical mechanics. We show that, unlike steady-state occupancy, DNA site exposure kinetics near a barrier is dominated by DNA sequence rather than by proximity to the barrier itself. The timescale for formation of positioning patterns near barriers is proportional to the timescale for active nucleosome eviction. We also show that there are strong gene-to-gene variations in nucleosome positioning near barriers, which are eliminated by averaging over many genes. Our results suggest that measurement of nucleosome kinetics can reveal information about sequence-dependent regulation that is not apparent in steady-state nucleosome occupancy. DOI: 10.1093/nar/gkt854 PMCID: PMC3874171 PMID: 24068556 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/17198702
1. Exp Cell Res. 2007 Feb 1;313(3):560-71. doi: 10.1016/j.yexcr.2006.10.033. Epub 2006 Nov 11. RhoB and the mammalian Diaphanous-related formin mDia2 in endosome trafficking. Wallar BJ(1), Deward AD, Resau JH, Alberts AS. Author information: (1)Laboratory of Cell Structure and Signal Integration, Van Andel Research Institute, 333 Bostwick Avenue, Grand Rapids, MI 49503, USA. Rho GTPases and the dynamic assembly and disassembly of actin filaments have been shown to have critical roles in both the internalization and trafficking of growth factor receptors. While all three mammalian Diaphanous-related (mDia1/2/3) formin GTPase effector proteins have been localized on endosomes, a role for their actin nucleation, filament elongation, and/or bundling remains poorly understood in the context of intracellular trafficking. In a study of a functional relationship between RhoB, a GTPase known to associate with both early- and late-endosomes, and the formin mDia2, we show that 1) RhoB and mDia2 interact on endosomes; 2) GTPase activity-the ability to hydrolyze GTP to GDP-is required for the ability of RhoB to govern endosome dynamics; and 3) the actin dynamics controlled by RhoB and mDia2 is necessary for vesicle trafficking. These studies further suggest that Rho GTPases significantly influence the activity of mDia family formins in driving cellular membrane remodeling through the regulation of actin dynamics. DOI: 10.1016/j.yexcr.2006.10.033 PMID: 17198702 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23060957
1. Commun Integr Biol. 2012 Jul 1;5(4):340-4. doi: 10.4161/cib.20214. mDia1-3 in mammalian filopodia. Goh WI(1), Ahmed S. Author information: (1)Institute of Medical Biology; Singapore; Immunos, Singapore. mDia proteins are members of the formin family of actin nucleating proteins that polymerize linear actin filaments. Such filaments form the core of thin, tubular, membrane-bound cell surface protrusions known as filopodia, which are a major feature of mammalian cell morphology. Filopodia are dynamic structures that help cells sense environmental cues, and play a role in cell migration, axon guidance, angiogenesis and other processes. RhoGTPases bind to and control the activity of mDia proteins, and several other binding partners of the three mDia1 isoforms-mDia1, mDia2 and mDia3-have been documented. Two independent pathways controlling mammalian filopodium formation have emerged, with one driven by the RhoGTPase Cdc42, and the other by Rif. While mDia2 has been the main formin implicated in forming filopodia, mDia1 has recently surfaced as the key formin utilized by both the Cdc42 and Rif pathways to drive filopodial protrusion. DOI: 10.4161/cib.20214 PMCID: PMC3460838 PMID: 23060957
http://www.ncbi.nlm.nih.gov/pubmed/18239683
1. EMBO J. 2008 Feb 20;27(4):618-28. doi: 10.1038/emboj.2008.7. Epub 2008 Jan 31. The mammalian formin FHOD1 is activated through phosphorylation by ROCK and mediates thrombin-induced stress fibre formation in endothelial cells. Takeya R(1), Taniguchi K, Narumiya S, Sumimoto H. Author information: (1)Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan. Formin-family proteins, in the active state, form actin-based structures such as stress fibres. Their activation mechanisms, however, are largely unknown except that mDia and its closely related formins can be activated by direct binding of the small GTPase Rho or Cdc42. Here we show that the Rho-dependent protein kinase ROCK phosphorylates the C-terminal residues Ser1131, Ser1137, and Thr1141 of formin homology domain protein 1 (FHOD1), a major endothelial formin that is normally autoinhibited by intramolecular interaction between the N- and C-terminal regions. Phosphorylation of FHOD1 at the three residues fully disrupts the autoinhibitory interaction, which culminates in formation of stress fibres. We also demonstrate that, in vascular endothelial cells, thrombin, a vasoactive substance leading to Rho activation, elicits both FHOD1 phosphorylation and stress fibre formation in a ROCK-dependent manner, and that FHOD1 depletion by RNA interference impairs thrombin-induced stress fibre formation. Based on these findings we propose a novel mechanism for activation of formin-family proteins: ROCK, activated by G protein-coupled receptor ligands such as thrombin, directly phosphorylates FHOD1 at the C-terminal region, which renders this formin in the active form, leading to stress fibre formation. DOI: 10.1038/emboj.2008.7 PMCID: PMC2262041 PMID: 18239683 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/26366214
1. J Cancer. 2015 Aug 22;6(10):1005-10. doi: 10.7150/jca.12703. eCollection 2015. Akt and Hippo Pathways in Ewing's Sarcoma Tumors and Their Prognostic Significance. Ahmed AA(1), Abedalthagafi M(1), Anwar AE(2), Bui MM(3). Author information: (1)1. Department of Pathology, King Fahad Medical City, Riyadh, Saudi Arabia. (2)2. Department of Epidemiology and Biostatistics, College of Public Health and Health Informatics, King Saud Bin Abdulaziz University for Health Sciences, Riyadh, Saudi Arabia. (3)3. Department of Anatomic Pathology, Moffitt Cancer Center/University of South Florida, Tampa, Florida, United States. BACKGROUND: Ewing's sarcoma tumor is an aggressive malignancy of bone and soft tissue in children and young adults. Despite advances in modern therapy, metastasis occurs and results in high mortality. Intracellular molecules Yap, Akt, mTOR, and Erk are signaling pathway members that regulate the proliferation of tumor cells. OBJECTIVE AND METHODS: We studied the immunohistochemical expression of these proteins in 36 tumor samples from adult and pediatric patients with Ewing's sarcoma tumors. Patients' age, sex, tumor site, tumor size, clinical stage and survival (overall and disease-free survival) were collected. Tissue microarrays slides were stained with antibodies against Yap, Akt, mTOR, and Erk proteins. RESULTS: Tumors exhibited variable expression of Yap, Akt, mTOR, and Erk (from negative, low to high), with high levels of expression present in 31%, 53%, 77% and 0% respectively. Immunohistochemical expression of Akt was associated with worse overall and disease-free survival (p<0.05). The other biomarkers did not demonstrate any difference in survival between low versus high expression. CONCLUSION: Although Yap, Akt, mTOR, and Erk protein are all expressed in Ewing's sarcoma by immunohistochemistry, only Akt expression is associated with worse prognosis. Larger studies are needed to verify these results and plan targeted therapy, particularly against Akt. DOI: 10.7150/jca.12703 PMCID: PMC4565850 PMID: 26366214 Conflict of interest statement: Conflicts of Interest: None.
http://www.ncbi.nlm.nih.gov/pubmed/18287523
1. Mol Biol Cell. 2008 May;19(5):2328-38. doi: 10.1091/mbc.e07-10-1086. Epub 2008 Feb 20. mDia2 induces the actin scaffold for the contractile ring and stabilizes its position during cytokinesis in NIH 3T3 cells. Watanabe S(1), Ando Y, Yasuda S, Hosoya H, Watanabe N, Ishizaki T, Narumiya S. Author information: (1)Department of Pharmacology, Kyoto University Faculty of Medicine, Kyoto 606-8501, Japan. mDia proteins are mammalian homologues of Drosophila diaphanous and belong to the formin family proteins that catalyze actin nucleation and polymerization. Although formin family proteins of nonmammalian species such as Drosophila diaphanous are essential in cytokinesis, whether and how mDia proteins function in cytokinesis remain unknown. Here we depleted each of the three mDia isoforms in NIH 3T3 cells by RNA interference and examined this issue. Depletion of mDia2 selectively increased the number of binucleate cells, which was corrected by coexpression of RNAi-resistant full-length mDia2. mDia2 accumulates in the cleavage furrow during anaphase to telophase, and concentrates in the midbody at the end of cytokinesis. Depletion of mDia2 induced contraction at aberrant sites of dividing cells, where contractile ring components such as RhoA, myosin, anillin, and phosphorylated ERM accumulated. Treatment with blebbistatin suppressed abnormal contraction, corrected localization of the above components, and revealed that the amount of F-actin at the equatorial region during anaphase/telophase was significantly decreased with mDia2 RNAi. These results demonstrate that mDia2 is essential in mammalian cell cytokinesis and that mDia2-induced F-actin forms a scaffold for the contractile ring and maintains its position in the middle of a dividing cell. DOI: 10.1091/mbc.e07-10-1086 PMCID: PMC2366861 PMID: 18287523 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/26039999
1. Proc Natl Acad Sci U S A. 2015 Jun 9;112(23):7255-60. doi: 10.1073/pnas.1505917112. Epub 2015 May 26. Netrin-1 exerts oncogenic activities through enhancing Yes-associated protein stability. Qi Q(1), Li DY(2), Luo HR(3), Guan KL(4), Ye K(5). Author information: (1)Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA 30322; (2)Program in Molecular Medicine, University of Utah, Salt Lake City, UT 84132; (3)Department of Pathology and Lab Medicine, Harvard Medical School and Children's Hospital Boston, Boston, MA 02115; (4)Department of Pharmacology and Moores Cancer Center, University of California at San Diego, La Jolla, CA 92037. (5)Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA 30322; kye@emory.edu. Yes-associated protein (YAP), a transcription coactivator, is the major downstream effector of the Hippo pathway, which plays a critical role in organ size control and cancer development. However, how YAP is regulated by extracellular stimuli in tumorigenesis remains incompletely understood. Netrin-1, a laminin-related secreted protein, displays proto-oncogenic activity in cancers. Nonetheless, the downstream signaling mediating its oncogenic effects is not well defined. Here we show that netrin-1 via its transmembrane receptors, deleted in colorectal cancer and uncoordinated-5 homolog, up-regulates YAP expression, escalating YAP levels in the nucleus and promoting cancer cell proliferation and migration. Inactivating netrin-1, deleted in colorectal cancer, or uncoordinated-5 homolog B (UNC5B) decreases YAP protein levels, abrogating cancer cell progression by netrin-1, whereas knockdown of mammalian STE20-like protein kinase 1/2 (MST1/2) or large tumor suppressor kinase 1/2 (Lats1/2), two sets of upstream core kinases of the Hippo pathway, has no effect in blocking netrin-1-induced up-regulation of YAP. Netrin-1 stimulates phosphatase 1A to dephosphorylate YAP, which leads to decreased ubiquitination and degradation, enhancing YAP accumulation and signaling. Hence, our findings support that netrin-1 exerts oncogenic activity through YAP signaling, providing a mechanism coupling extracellular signals to the nuclear YAP oncogene. DOI: 10.1073/pnas.1505917112 PMCID: PMC4466726 PMID: 26039999 [Indexed for MEDLINE] Conflict of interest statement: The authors declare no conflict of interest.
http://www.ncbi.nlm.nih.gov/pubmed/20513433
1. Mol Cell. 2010 May 28;38(4):590-602. doi: 10.1016/j.molcel.2010.02.040. SWI/SNF has intrinsic nucleosome disassembly activity that is dependent on adjacent nucleosomes. Dechassa ML(1), Sabri A, Pondugula S, Kassabov SR, Chatterjee N, Kladde MP, Bartholomew B. Author information: (1)Department of Biochemistry and Molecular Biology, Southern Illinois University School of Medicine, Carbondale, IL 62901-4413, USA. Comment in Mol Cell. 2010 May 28;38(4):484-6. doi: 10.1016/j.molcel.2010.05.005. The ATP-dependent chromatin remodeling complex SWI/SNF regulates transcription and has been implicated in promoter nucleosome eviction. Efficient nucleosome disassembly by SWI/SNF alone in biochemical assays, however, has not been directly observed. Employing a model system of dinucleosomes rather than mononucleosomes, we demonstrate that remodeling leads to ordered and efficient disassembly of one of the two nucleosomes. An H2A/H2B dimer is first rapidly displaced, and then, in a slower reaction, an entire histone octamer is lost. Nucleosome disassembly by SWI/SNF did not require additional factors such as chaperones or acceptors of histones. Observations in single molecules as well as bulk measurement suggest that a key intermediate in this process is one in which a nucleosome is moved toward the adjacent nucleosome. SWI/SNF recruited by the transcriptional activator Gal4-VP16 preferentially mobilizes the proximal nucleosome and destabilizes the adjacent nucleosome. Copyright 2010 Elsevier Inc. All rights reserved. DOI: 10.1016/j.molcel.2010.02.040 PMCID: PMC3161732 PMID: 20513433 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/17235287
1. EMBO J. 2007 Feb 7;26(3):730-40. doi: 10.1038/sj.emboj.7601524. Epub 2007 Jan 18. Activation domains drive nucleosome eviction by SWI/SNF. Gutiérrez JL(1), Chandy M, Carrozza MJ, Workman JL. Author information: (1)Stowers Institute for Medical Research, Kansas City, MO 64110, USA. ATP-dependent chromatin remodeling complexes play a critical role in chromatin dynamics. A large number of in vitro studies have pointed towards nucleosome sliding as the principal remodeling outcome of SWI/SNF action, whereas few have described histone octamer transfer as the principal outcome. In contrast, recent in vivo studies have linked the activity of SWI/SNF to histone eviction in trans from gene promoters. In this study, we have found that the chimeric transcription factor Gal4-VP16 can enhance SWI/SNF histone octamer transfer activity, resulting in targeted histone eviction from a nucleosome probe. This effect is dependent on the presence of the activation domain. We observed that under conditions mimicking the in vivo relative abundance of SWI/SNF with respect to the total number of nucleosomes in a cell nucleus, the accessibility of the transcription factor binding site is the first determinant in the sequence of events leading to nucleosome remodeling. We propose a model mechanism for this transcription factor-mediated enhancement of SWI/SNF octamer transfer activity. DOI: 10.1038/sj.emboj.7601524 PMCID: PMC1794382 PMID: 17235287 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19470761
1. Mol Cell Biol. 2009 Aug;29(15):4220-34. doi: 10.1128/MCB.01882-08. Epub 2009 May 26. An rtt109-independent role for vps75 in transcription-associated nucleosome dynamics. Selth LA(1), Lorch Y, Ocampo-Hafalla MT, Mitter R, Shales M, Krogan NJ, Kornberg RD, Svejstrup JQ. Author information: (1)Clare Hall Laboratories, Cancer Research UK London Research Institute, Blanche Lane, South Mimms, Hertfordshire, U.K. The histone chaperone Vps75 forms a complex with, and stimulates the activity of, the histone acetyltransferase Rtt109. However, Vps75 can also be isolated on its own and might therefore possess Rtt109-independent functions. Analysis of epistatic miniarray profiles showed that VPS75 genetically interacts with factors involved in transcription regulation whereas RTT109 clusters with genes linked to DNA replication/repair. Additional genetic and biochemical experiments revealed a close relationship between Vps75 and RNA polymerase II. Furthermore, Vps75 is recruited to activated genes in an Rtt109-independent manner, and its genome-wide association with genes correlates with transcription rate. Expression microarray analysis identified a number of genes whose normal expression depends on VPS75. Interestingly, histone H2B dynamics at some of these genes are consistent with a role for Vps75 in histone H2A/H2B eviction/deposition during transcription. Indeed, reconstitution of nucleosome disassembly using the ATP-dependent chromatin remodeler Rsc and Vps75 revealed that these proteins can cooperate to remove H2A/H2B dimers from nucleosomes. These results indicate a role for Vps75 in nucleosome dynamics during transcription, and importantly, this function appears to be largely independent of Rtt109. DOI: 10.1128/MCB.01882-08 PMCID: PMC2715805 PMID: 19470761 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23150908
1. N Engl J Med. 2013 Jan 10;368(2):107-16. doi: 10.1056/NEJMoa1211103. Epub 2012 Nov 14. Variant of TREM2 associated with the risk of Alzheimer's disease. Jonsson T(1), Stefansson H, Steinberg S, Jonsdottir I, Jonsson PV, Snaedal J, Bjornsson S, Huttenlocher J, Levey AI, Lah JJ, Rujescu D, Hampel H, Giegling I, Andreassen OA, Engedal K, Ulstein I, Djurovic S, Ibrahim-Verbaas C, Hofman A, Ikram MA, van Duijn CM, Thorsteinsdottir U, Kong A, Stefansson K. Author information: (1)deCODE Genetics, Reykjavik, Iceland. Comment in N Engl J Med. 2013 Jan 10;368(2):182-4. doi: 10.1056/NEJMe1213157. Nat Rev Neurol. 2013 Jan;9(1):5. doi: 10.1038/nrneurol.2012.254. Clin Genet. 2013 Jun;83(6):525-6. doi: 10.1111/cge.12108. N Engl J Med. 2013 Oct 17;369(16):1568-9. doi: 10.1056/NEJMc1306509. N Engl J Med. 2013 Oct 17;369(16):1564-5. doi: 10.1056/NEJMc1306509. N Engl J Med. 2013 Oct 17;369(16):1565. doi: 10.1056/NEJMc1306509. N Engl J Med. 2013 Oct 17;369(16):1568. doi: 10.1056/NEJMc1306509. BACKGROUND: Sequence variants, including the ε4 allele of apolipoprotein E, have been associated with the risk of the common late-onset form of Alzheimer's disease. Few rare variants affecting the risk of late-onset Alzheimer's disease have been found. METHODS: We obtained the genome sequences of 2261 Icelanders and identified sequence variants that were likely to affect protein function. We imputed these variants into the genomes of patients with Alzheimer's disease and control participants and then tested for an association with Alzheimer's disease. We performed replication tests using case-control series from the United States, Norway, The Netherlands, and Germany. We also tested for a genetic association with cognitive function in a population of unaffected elderly persons. RESULTS: A rare missense mutation (rs75932628-T) in the gene encoding the triggering receptor expressed on myeloid cells 2 (TREM2), which was predicted to result in an R47H substitution, was found to confer a significant risk of Alzheimer's disease in Iceland (odds ratio, 2.92; 95% confidence interval [CI], 2.09 to 4.09; P=3.42×10(-10)). The mutation had a frequency of 0.46% in controls 85 years of age or older. We observed the association in additional sample sets (odds ratio, 2.90; 95% CI, 2.16 to 3.91; P=2.1×10(-12) in combined discovery and replication samples). We also found that carriers of rs75932628-T between the ages of 80 and 100 years without Alzheimer's disease had poorer cognitive function than noncarriers (P=0.003). CONCLUSIONS: Our findings strongly implicate variant TREM2 in the pathogenesis of Alzheimer's disease. Given the reported antiinflammatory role of TREM2 in the brain, the R47H substitution may lead to an increased predisposition to Alzheimer's disease through impaired containment of inflammatory processes. (Funded by the National Institute on Aging and others.). DOI: 10.1056/NEJMoa1211103 PMCID: PMC3677583 PMID: 23150908 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/17088018
1. Neurosci Lett. 2007 Jan 10;411(2):133-7. doi: 10.1016/j.neulet.2006.10.029. Epub 2006 Nov 7. Absence of TREM2 polymorphisms in patients with Alzheimer's disease and Frontotemporal Lobar Degeneration. Fenoglio C(1), Galimberti D, Piccio L, Scalabrini D, Panina P, Buonsanti C, Venturelli E, Lovati C, Forloni G, Mariani C, Bresolin N, Scarpini E. Author information: (1)Department of Neurological Sciences, "Dino Ferrari" Center, University of Milan, IRCCS Fondazione Ospedale Maggiore Policlinico, Milan, Italy. Triggering Receptor Expressed on Myeloid cells (TREM)2 deficiency originates a genetic syndrome characterized by bone cysts and presenile dementia, named Nasu-Hakola disease (NHD). Early onset dementia and marked involvement of frontal regions are features characterizing both NHD and other kinds of neurodegenerative disorders, such as Frontotemporal Lobar Degeneration (FTLD), and, in some cases, Alzheimer's disease (AD). Three Single Nucleotide Polymorphisms (SNPs) in TREM2 coding region were screened by allelic discrimination in a population of probable AD patients as well as FTLD patients as compared with age-matched controls. In addition, mutation scanning of the coding region of TREM2 gene was carried out in 7 patients with early onset AD (EOAD), 16 FTLD, and 20 controls. None of the SNPs analyzed was present, either in patients or controls. Moreover, mutation scanning of the five exons of TREM2 failed to detect the presence of novel polymorphisms. These data demonstrate that TREM2 coding region is highly conserved, implying a crucial role of this receptor. Further studies, including a functional analysis, are certainly required to clarify the role of TREM2 in neurodegenerative processes. DOI: 10.1016/j.neulet.2006.10.029 PMID: 17088018 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19029894
1. Nat Struct Mol Biol. 2008 Dec;15(12):1272-7. doi: 10.1038/nsmb.1524. Epub 2008 Nov 23. Structure of a RSC-nucleosome complex and insights into chromatin remodeling. Chaban Y(1), Ezeokonkwo C, Chung WH, Zhang F, Kornberg RD, Maier-Davis B, Lorch Y, Asturias FJ. Author information: (1)Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA. ATP-dependent chromatin-remodeling complexes, such as RSC, can reposition, evict or restructure nucleosomes. A structure of a RSC-nucleosome complex with a nucleosome determined by cryo-EM shows the nucleosome bound in a central RSC cavity. Extensive interaction of RSC with histones and DNA seems to destabilize the nucleosome and lead to an overall ATP-independent rearrangement of its structure. Nucleosomal DNA appears disordered and largely free to bulge out into solution as required for remodeling, but the structure of the RSC-nucleosome complex indicates that RSC is unlikely to displace the octamer from the nucleosome to which it is bound. Consideration of the RSC-nucleosome structure and published biochemical information suggests that ATP-dependent DNA translocation by RSC may result in the eviction of histone octamers from adjacent nucleosomes. DOI: 10.1038/nsmb.1524 PMCID: PMC2659406 PMID: 19029894 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/15046306
1. Syst Appl Microbiol. 2004 Mar;27(2):175-85. doi: 10.1078/072320204322881790. Use of the genomic signature in bacterial classification and identification. Coenye T(1), Vandamme P. Author information: (1)Laboratorium voor Microbiologie, Ghent University, Gent, Belgium. Tom.Coenye@UGent.be In this study we investigated the correlation between dinucleotide relative abundance values (the genomic signature) obtained from bacterial whole-genome sequences and two parameters widely used for bacterial classification, 16S rDNA sequence similarity and DNA-DNA hybridisation values. Twenty-eight completely sequenced bacterial genomes were included in the study. The correlation between the genomic signature and DNA-DNA hybridisation values was high and taxa that showed less than 30% DNA-DNA binding will in general not have dinucleotide relative abundance dissimilarity (delta*) values below 40. On the other hand, taxa showing more than 50% DNA-DNA binding will not have delta* values higher than 17. Our data indicate that the overall correlation between genomic signature and 16S rDNA sequence similarity is low, except for closely related organisms (16S rDNA similarity >94%). Statistical analysis of delta* values between different subgroups of the Proteobacteria indicate that the beta- and gamma-Proteobacteria are more closely related to each other than to the other subgroups of the Proteobacteria and that the alpha- and epsilon-Proteobacteria form clearly separate subgroups. Using the genomic signature we have also predicted DNA-DNA binding values for fastidious or unculturable endosymbionts belonging to the genera Rickettsia, Wigglesworthia and Buchnera. DOI: 10.1078/072320204322881790 PMID: 15046306 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/9190805
1. J Bacteriol. 1997 Jun;179(12):3899-913. doi: 10.1128/jb.179.12.3899-3913.1997. Compositional biases of bacterial genomes and evolutionary implications. Karlin S(1), Mrázek J, Campbell AM. Author information: (1)Department of Mathematics, Stanford University, California 94305-2125, USA. We compare and contrast genome-wide compositional biases and distributions of short oligonucleotides across 15 diverse prokaryotes that have substantial genomic sequence collections. These include seven complete genomes (Escherichia coli, Haemophilus influenzae, Mycoplasma genitalium, Mycoplasma pneumoniae, Synechocystis sp. strain PCC6803, Methanococcus jannaschii, and Pyrobaculum aerophilum). A key observation concerns the constancy of the dinucleotide relative abundance profiles over multiple 50-kb disjoint contigs within the same genome. (The profile is rhoXY* = fXY*/fX*fY* for all XY, where fX* denotes the frequency of the nucleotide X and fY* denotes the frequency of the dinucleotide XY, both computed from the sequence concatenated with its inverted complementary sequence.) On the basis of this constancy, we refer to the collection [rhoXY*] as the genome signature. We establish that the differences between [rhoXY*] vectors of 50-kb sample contigs of different genomes virtually always exceed the differences between those of the same genomes. Various di- and tetranucleotide biases are identified. In particular, we find that the dinucleotide CpG=CG is underrepresented in many thermophiles (e.g., M. jannaschii, Sulfolobus sp., and M. thermoautotrophicum) but overrepresented in halobacteria. TA is broadly underrepresented in prokaryotes and eukaryotes, but normal counts appear in Sulfolobus and P. aerophilum sequences. More than for any other bacterial genome, palindromic tetranucleotides are underrepresented in H. influenzae. The M. jannaschii sequence is unprecedented in its extreme underrepresentation of CTAG tetranucleotides and in the anomalous distribution of CTAG sites around the genome. Comparative analysis of numbers of long tetranucleotide microsatellites distinguishes H. influenzae. Dinucleotide relative abundance differences between bacterial sequences are compared. For example, in these assessments of differences, the cyanobacteria Synechocystis, Synechococcus, and Anabaena do not form a coherent group and are as far from each other as general gram-negative sequences are from general gram-positive sequences. The difference of M. jannaschii from low-G+C gram-positive proteobacteria is one-half of the difference from gram-negative proteobacteria. Interpretations and hypotheses center on the role of the genome signature in highlighting similarities and dissimilarities across different classes of prokaryotic species, possible mechanisms underlying the genome signature, the form and level of genome compositional flux, the use of the genome signature as a chronometer of molecular phylogeny, and implications with respect to the three putative eubacterial, archaeal, and eukaryote domains of life and to the origin and early evolution of eukaryotes. DOI: 10.1128/jb.179.12.3899-3913.1997 PMCID: PMC179198 PMID: 9190805 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/9294192
1. Proc Natl Acad Sci U S A. 1997 Sep 16;94(19):10227-32. doi: 10.1073/pnas.94.19.10227. Compositional differences within and between eukaryotic genomes. Karlin S(1), Mrázek J. Author information: (1)Department of Mathematics, Stanford University, Stanford, CA 94305-2125, USA. Eukaryotic genome similarity relationships are inferred using sequence information derived from large aggregates of genomic sequences. Comparisons within and between species sample sequences are based on the profile of dinucleotide relative abundance values (The profile is rho*XY = f*XY/f*Xf*Y for all XY, where f*X denotes the frequency of the nucleotide X and f*XY denotes the frequency of the dinucleotide XY, both computed from the sequence concatenated with its inverted complement). Previous studies with respect to prokaryotes and this study document that profiles of different DNA sequence samples (sample size >/=50 kb) from the same organism are generally much more similar to each other than they are to profiles from other organisms, and that closely related organisms generally have more similar profiles than do distantly related organisms. On this basis we refer to the collection (rho*XY) as the genome signature. This paper identifies rho*XY extremes and compares genome signature differences for a diverse range of eukaryotic species. Interpretations on the mechanisms maintaining these profile differences center on genome-wide replication, repair, DNA structures, and context-dependent mutational biases. It is also observed that mitochondrial genome signature differences between species parallel the corresponding nuclear genome signature differences despite large differences between corresponding mitochondrial and nuclear signatures. The genome signature differences also have implications for contrasts between rodents and other mammals, and between monocot and dicot plants, as well as providing evidence for similarities among fungi and the diversity of protists. DOI: 10.1073/pnas.94.19.10227 PMCID: PMC23344 PMID: 9294192 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/15716010
1. Gene. 2005 Feb 14;346:173-85. doi: 10.1016/j.gene.2004.10.021. The spectrum of genomic signatures: from dinucleotides to chaos game representation. Wang Y(1), Hill K, Singh S, Kari L. Author information: (1)Department of Computer Science, University of Western Ontario, London, Ontario, Canada N6A 5B7. ywang@upei.ca In the post genomic era, access to complete genome sequence data for numerous diverse species has opened multiple avenues for examining and comparing primary DNA sequence organization of entire genomes. Previously, the concept of a genomic signature was introduced with the observation of species-type specific Dinucleotide Relative Abundance Profiles (DRAPs); dinucleotides were identified as the subsequences with the greatest bias in representation in a majority of genomes. Herein, we demonstrate that DRAP is one particular genomic signature contained within a broader spectrum of signatures. Within this spectrum, an alternative genomic signature, Chaos Game Representation (CGR), provides a unique visualization of patterns in sequence organization. A genomic signature is associated with a particular integer order or subsequence length that represents a measure of the resolution or granularity in the analysis of primary DNA sequence organization. We quantitatively explore the organizational information provided by genomic signatures of different orders through different distance measures, including a novel Image Distance. The Image Distance and other existing distance measures are evaluated by comparing the phylogenetic trees they generate for 26 complete mitochondrial genomes from a diversity of species. The phylogenetic tree generated by the Image Distance is compatible with the known relatedness of species. Quantitative evaluation of the spectrum of genomic signatures may be used to ultimately gain insight into the determinants and biological relevance of the genome signatures. DOI: 10.1016/j.gene.2004.10.021 PMID: 15716010 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/12171605
1. BMC Genomics. 2002 Aug 9;3(1):23. doi: 10.1186/1471-2164-3-23. Pervasive properties of the genomic signature. Jernigan RW(1), Baran RH. Author information: (1)Department of Mathematics and Statistics, The American University, Washington, DC 20016, USA. jernigan@american.edu BACKGROUND: The dinucleotide relative abundance profile can be regarded as a genomic signature because, despite diversity between species, it varies little between 50 kilobase or longer windows on a given genome. Both the causes and the functional significance of this phenomenon could be illuminated by determining if it persists on smaller scales. The profile is computed from the base step "odds ratios" that compare dinucleotide frequencies to those expected under the assumption of stochastic equilibrium (thorough shuffling). Analysis is carried out on 22 sequences, representing 19 species and comprised of about 53 million bases all together, to assess stability of the signature in windows ranging in size from 50 kilobases down to 125 bases. RESULTS: Dinucleotide relative abundance distance from the global signature is computed locally for all non-overlapping windows on each sequence. These distances are log-normally distributed with nearly constant variance and with means that tend to zero slower than reciprocal square root of window size. The mean distance within genomes is larger for protist, plant, and human chromosomes, and smaller for archaea, bacteria, and yeast, for any window size. CONCLUSIONS: The imprint of the global signature is locally pervasive on all scales considered in the sequences (either genomes or chromosomes) that were scanned. DOI: 10.1186/1471-2164-3-23 PMCID: PMC126251 PMID: 12171605
http://www.ncbi.nlm.nih.gov/pubmed/23407992
1. Mol Neurobiol. 2013 Aug;48(1):180-5. doi: 10.1007/s12035-013-8424-8. Epub 2013 Feb 14. TREM2 in Alzheimer's disease. Jiang T(1), Yu JT, Zhu XC, Tan L. Author information: (1)Department of Neurology, Qingdao Municipal Hospital, Nanjing Medical University, Nanjing, China. Recent works have demonstrated a rare functional variant (R47H) in triggering receptor expressed on myeloid cells (TREM) 2 gene, encoding TREM2 protein, increase susceptibility to late-onset Alzheimer's disease (AD), with an odds ratio similar to that of the apolipoprotein E ε4 allele. The reduced function of TREM2 was speculated to be the main cause in the pathogenic effects of this risk variant, and TREM2 is highly expressed in white matter, as well as in the hippocampus and neocortex, which is partly consistent with the pathological features reported in AD brain, indicating the possible involvement of TREM2 in AD pathogenesis. Emerging evidence has demonstrated that TREM2 could suppress inflammatory response by repression of microglia-mediated cytokine production and secretion, which may prevent inflammation-induced bystander damage of neurons. TREM2 also participates in the regulation of phagocytic pathways that are responsible for the removal of neuronal debris. In this article, we review the recent epidemiological findings of TREM2 that related with late-onset AD and speculate the possible roles of TREM2 in progression of this disease. Based on the potential protective actions of TREM2 in AD pathogenesis, targeting TREM2 might provide new opportunities for AD treatment. DOI: 10.1007/s12035-013-8424-8 PMID: 23407992 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/24008565
1. Nat Struct Mol Biol. 2013 Sep;20(9):1026-32. doi: 10.1038/nsmb.2648. Nucleosome sliding mechanisms: new twists in a looped history. Mueller-Planitz F(1), Klinker H, Becker PB. Author information: (1)1] Adolf-Butenandt-Institute, Ludwig-Maximilians-Universität, Munich, Germany. [2] Center for Integrated Protein Science Munich, Ludwig-Maximilians-Universität, Munich, Germany. Nucleosomes, the basic organizational units of chromatin, package and regulate eukaryotic genomes. ATP-dependent nucleosome-remodeling factors endow chromatin with structural flexibility by promoting assembly or disruption of nucleosomes and the exchange of histone variants. Furthermore, most remodeling factors induce nucleosome movements through sliding of histone octamers on DNA. We summarize recent progress toward unraveling the basic nucleosome sliding mechanism and the interplay of the remodelers' DNA translocases with accessory domains. Such domains optimize and regulate the basic sliding reaction and exploit sliding to achieve diverse structural effects, such as nucleosome positioning or eviction, or the regular spacing of nucleosomes in chromatin. DOI: 10.1038/nsmb.2648 PMID: 24008565 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/10430918
1. Proc Natl Acad Sci U S A. 1999 Aug 3;96(16):9190-5. doi: 10.1073/pnas.96.16.9190. A chimeric prokaryotic ancestry of mitochondria and primitive eukaryotes. Karlin S(1), Brocchieri L, Mrázek J, Campbell AM, Spormann AM. Author information: (1)Department of Mathematics, Stanford University, Stanford, CA 94305-2125, USA. We provide data and analysis to support the hypothesis that the ancestor of animal mitochondria (Mt) and many primitive amitochondrial (a-Mt) eukaryotes was a fusion microbe composed of a Clostridium-like eubacterium and a Sulfolobus-like archaebacterium. The analysis is based on several observations: (i) The genome signatures (dinucleotide relative abundance values) of Clostridium and Sulfolobus are compatible (sufficiently similar) and each has significantly more similarity in genome signatures with animal Mt sequences than do all other available prokaryotes. That stable fusions may require compatibility in genome signatures is suggested by the compatibility of plasmids and hosts. (ii) The expanded energy metabolism of the fusion organism was strongly selective for cementing such a fusion. (iii) The molecular apparatus of endospore formation in Clostridium serves as raw material for the development of the nucleus and cytoplasm of the eukaryotic cell. DOI: 10.1073/pnas.96.16.9190 PMCID: PMC17755 PMID: 10430918 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/18953039
1. Nucleic Acids Res. 2008 Dec;36(22):e147. doi: 10.1093/nar/gkn753. Epub 2008 Oct 25. Using Mahalanobis distance to compare genomic signatures between bacterial plasmids and chromosomes. Suzuki H(1), Sota M, Brown CJ, Top EM. Author information: (1)Department of Biological Sciences, University of Idaho, Moscow, ID 83844, USA. Plasmids are ubiquitous mobile elements that serve as a pool of many host beneficial traits such as antibiotic resistance in bacterial communities. To understand the importance of plasmids in horizontal gene transfer, we need to gain insight into the 'evolutionary history' of these plasmids, i.e. the range of hosts in which they have evolved. Since extensive data support the proposal that foreign DNA acquires the host's nucleotide composition during long-term residence, comparison of nucleotide composition of plasmids and chromosomes could shed light on a plasmid's evolutionary history. The average absolute dinucleotide relative abundance difference, termed delta-distance, has been commonly used to measure differences in dinucleotide composition, or 'genomic signature', between bacterial chromosomes and plasmids. Here, we introduce the Mahalanobis distance, which takes into account the variance-covariance structure of the chromosome signatures. We demonstrate that the Mahalanobis distance is better than the delta-distance at measuring genomic signature differences between plasmids and chromosomes of potential hosts. We illustrate the usefulness of this metric for proposing candidate long-term hosts for plasmids, focusing on the virulence plasmids pXO1 from Bacillus anthracis, and pO157 from Escherichia coli O157:H7, as well as the broad host range multi-drug resistance plasmid pB10 from an unknown host. DOI: 10.1093/nar/gkn753 PMCID: PMC2602791 PMID: 18953039 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/7809131
1. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12837-41. doi: 10.1073/pnas.91.26.12837. Heterogeneity of genomes: measures and values. Karlin S(1), Ladunga I, Blaisdell BE. Author information: (1)Department of Mathematics, Stanford University, CA 94305-2125. Genomic homogeneity is investigated for a broad base of DNA sequences in terms of dinucleotide relative abundance distances (abbreviated delta-distances) and of oligonucleotide compositional extremes. It is shown that delta-distances between different genomic sequences in the same species are low, only about 2 or 3 times the distance found in random DNA, and are generally smaller than the between-species delta-distances. Extremes in short oligonucleotides include underrepresentation of TpA and overrepresentation of GpC in most temperate bacteriophage sequences; underrepresentation of CTAG in most eubacterial genomes; underrepresentation of GATC in most bacteriophage; CpG suppression in vertebrates, in all animal mitochondrial genomes, and in many thermophilic bacterial sequences; and overrepresentation of GpG/CpC in all animal mitochondrial sets and chloroplast genomes. Interpretations center on DNA structures (dinucleotide stacking energies, DNA curvature and superhelicity, nucleosome organization), context-dependent mutational events, methylation effects, and processes of replication and repair. DOI: 10.1073/pnas.91.26.12837 PMCID: PMC45535 PMID: 7809131 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/10066522
1. Curr Opin Microbiol. 1998 Oct;1(5):598-610. doi: 10.1016/s1369-5274(98)80095-7. Global dinucleotide signatures and analysis of genomic heterogeneity. Karlin S(1). Author information: (1)Department of Mathematics, Stanford University, Stanford, CA 94305-2125, USA. Early biochemical experiments measuring nearest neighbor frequencies established that the set of dinucleotide relative abundance values (dinucleotide biases) is a remarkably stable property of the DNA of an organism. Analyses of currently available genomic sequence data have extended these earlier results, showing that the dinucleotide biases evaluated for successive 50 kb segments of a genome are significantly more similar to each other than to those of sequences from more distant organisms. From this perspective, the set of dinucleotide biases constitutes a 'genomic signature' that can discriminate sequences from different organisms. The dinucleotide biases appear to reflect species-specific properties of DNA stacking energies, modification, replication, and repair mechanisms. The genomic signature is useful for detecting pathogenicity islands in bacterial genomes. DOI: 10.1016/s1369-5274(98)80095-7 PMID: 10066522 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19933844
1. Mol Cell Biol. 2010 Feb;30(3):657-74. doi: 10.1128/MCB.01117-09. Epub 2009 Nov 23. Fission yeast Iec1-ino80-mediated nucleosome eviction regulates nucleotide and phosphate metabolism. Hogan CJ(1), Aligianni S, Durand-Dubief M, Persson J, Will WR, Webster J, Wheeler L, Mathews CK, Elderkin S, Oxley D, Ekwall K, Varga-Weisz PD. Author information: (1)Chromatin and Gene Expression, Babraham Institute, Babraham, Cambridge CB22 3AT, UK. Ino80 is an ATP-dependent nucleosome-remodeling enzyme involved in transcription, replication, and the DNA damage response. Here, we characterize the fission yeast Ino80 and find that it is essential for cell viability. We show that the Ino80 complex from fission yeast mediates ATP-dependent nucleosome remodeling in vitro. The purification of the Ino80-associated complex identified a highly conserved complex and the presence of a novel zinc finger protein with similarities to the mammalian transcriptional regulator Yin Yang 1 (YY1) and other members of the GLI-Krüppel family of proteins. Deletion of this Iec1 protein or the Ino80 complex subunit arp8, ies6, or ies2 causes defects in DNA damage repair, the response to replication stress, and nucleotide metabolism. We show that Iec1 is important for the correct expression of genes involved in nucleotide metabolism, including the ribonucleotide reductase subunit cdc22 and phosphate- and adenine-responsive genes. We find that Ino80 is recruited to a large number of promoter regions on phosphate starvation, including those of phosphate- and adenine-responsive genes that depend on Iec1 for correct expression. Iec1 is required for the binding of Ino80 to target genes and subsequent histone loss at the promoter and throughout the body of these genes on phosphate starvation. This suggests that the Iec1-Ino80 complex promotes transcription through nucleosome eviction. DOI: 10.1128/MCB.01117-09 PMCID: PMC2812234 PMID: 19933844 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/18836177
1. Mol Cell Proteomics. 2009 Feb;8(2):258-72. doi: 10.1074/mcp.M800060-MCP200. Epub 2008 Oct 3. Analysis of protein processing by N-terminal proteomics reveals novel species-specific substrate determinants of granzyme B orthologs. Van Damme P(1), Maurer-Stroh S, Plasman K, Van Durme J, Colaert N, Timmerman E, De Bock PJ, Goethals M, Rousseau F, Schymkowitz J, Vandekerckhove J, Gevaert K. Author information: (1)Department of Medical Protein Research, Flanders Institute for Biotechnology (VIB), Ghent, Belgium. Using a targeted peptide-centric proteomics approach, we performed in vitro protease substrate profiling of the apoptotic serine protease granzyme B resulting in the delineation of more than 800 cleavage sites in 322 human and 282 mouse substrates, encompassing the known substrates Bid, caspase-7, lupus La protein, and fibrillarin. Triple SILAC (stable isotope labeling by amino acids in cell culture) further permitted intra-experimental evaluation of species-specific variations in substrate selection by the mouse or human granzyme B ortholog. For the first time granzyme B substrate specificities were directly mapped on a proteomic scale and revealed unknown cleavage specificities, uncharacterized extended specificity profiles, and macromolecular determinants in substrate selection that were confirmed by molecular modeling. We further tackled a substrate hunt in an in vivo setup of natural killer cell-mediated cell death confirming in vitro characterized granzyme B cleavages next to several other unique and hitherto unreported proteolytic events in target cells. DOI: 10.1074/mcp.M800060-MCP200 PMID: 18836177 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23294434
1. Curr Comput Aided Drug Des. 2013 Mar;9(1):108-17. Biomedical data integration in computational drug design and bioinformatics. Seoane JA(1), Aguiar-Pulido V, Munteanu CR, Rivero D, Rabunal JR, Dorado J, Pazos A. Author information: (1)Department of Information and Communication Technologies, Computer Science School, University of A Coruña, Spain. In recent years, in the post genomic era, more and more data is being generated by biological high throughput technologies, such as proteomics and transcriptomics. This omics data can be very useful, but the real challenge is to analyze all this data, as a whole, after integrating it. Biomedical data integration enables making queries to different, heterogeneous and distributed biomedical data sources. Data integration solutions can be very useful not only in the context of drug design, but also in biomedical information retrieval, clinical diagnosis, system biology, etc. In this review, we analyze the most common approaches to biomedical data integration, such as federated databases, data warehousing, multi-agent systems and semantic technology, as well as the solutions developed using these approaches in the past few years. PMID: 23294434 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23964590
1. J Proteome Res. 2013 Sep 6;12(9):3823-30. doi: 10.1021/pr400435d. Epub 2013 Aug 21. Novel highly sensitive, specific, and straightforward strategy for comprehensive N-terminal proteomics reveals unknown substrates of the mitochondrial peptidase Icp55. Venne AS(1), Vögtle FN, Meisinger C, Sickmann A, Zahedi RP. Author information: (1)Leibniz-Institut für Analytische Wissenschaften-ISAS-e.V., Dortmund, Germany. We present a novel straightforward method for enrichment of N-terminal peptides, utilizing charge-based fractional diagonal chromatography (ChaFRADIC). Our method is robust, easy to operate, fast, specific, and more sensitive than existing methods, enabling the differential quantitation of 1459 nonredundant N-terminal peptides between two S. cerevisiae samples within 10 h of LC-MS, starting from only 50 μg of protein per condition and analyzing only 40% of the obtained fractions. Using ChaFRADIC we compared mitochondrial proteins from wild-type and icp55Δ yeast (30 μg each). Icp55 is an intermediate cleaving peptidase, which, following mitochondrial processing peptidase (MPP)-dependent cleavage of signal sequences, removes a single amino acid from a specific set of proteins according to the N-end rule. Using ChaFRADIC we identified 36 icp55 substrates, 14 of which were previously unknown, expanding the set of known icp55 substrates to a total of 52 proteins. Interestingly, a novel substrate, Isa2, is likely processed by Icp55 in two consecutive steps and thus might represent the first example of a triple processing event in a mitochondrial precursor protein. Thus, ChaFRADIC is a powerful and practicable tool for protease and peptidase research, providing the sensitivity to characterize even samples that can be obtained only in small quantities. DOI: 10.1021/pr400435d PMID: 23964590 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23287290
1. Curr Opin Chem Biol. 2013 Feb;17(1):34-40. doi: 10.1016/j.cbpa.2012.12.007. Epub 2013 Jan 1. Glycomics meets genomics, epigenomics and other high throughput omics for system biology studies. Zoldoš V(1), Horvat T, Lauc G. Author information: (1)University of Zagreb, Faculty of Science, Zagreb, Croatia. Majority of eukaryotic proteins are glycosylated and their glycan moieties have numerous important structural, functional and regulatory roles. Because of structural complexity of glycans and technological limitations glycomics, and particularly glycoproteomics was not able to follow rapid progress in genomics and proteomics over last 30 years. However, the field of glycan has been progressing rapidly and first large-scale studies of the glycome have been completed recently. These studies have revealed significant differences in glycome composition between individuals, which may contribute to the human phenotypic variability. The current state-of-the-art in high-throughput glycomics and its integration with genomics, epigenomics and lipidomics is reviewed in this article. Copyright © 2012 Elsevier Ltd. All rights reserved. DOI: 10.1016/j.cbpa.2012.12.007 PMID: 23287290 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/26199863
1. FEBS Open Bio. 2015 Jun 22;5:542-9. doi: 10.1016/j.fob.2015.06.007. eCollection 2015. Small molecules inhibiting the nuclear localization of YAP/TAZ for chemotherapeutics and chemosensitizers against breast cancers. Oku Y(1), Nishiya N(1), Shito T(1), Yamamoto R(1), Yamamoto Y(1), Oyama C(1), Uehara Y(1). Author information: (1)Department of Microbial Chemical Biology and Drug Discovery, Iwate Medical University School of Pharmaceutical Sciences, 2-1-1 Nishitokuta, Yahaba-cho, Shiwa-gun, Iwate 028-3694, Japan. YAP and TAZ oncoproteins confer malignancy and drug resistance to various cancer types. We screened for small molecules that inhibit the nuclear localization of YAP/TAZ. Dasatinib, statins and pazopanib inhibited the nuclear localization and target gene expression of YAP and TAZ. All three drugs induced phosphorylation of YAP and TAZ, and pazopanib induced proteasomal degradation of YAP/TAZ. The sensitivities to these drugs are correlated with dependence on YAP/TAZ in breast cancer cell lines. Combinations of these compounds with each other or with other anti-cancer drugs efficiently reduced cell proliferation of YAP/TAZ-dependent breast cancer cells. These results suggest that these drugs can be therapeutics and chemosensitizers for YAP/TAZ-dependent breast cancers. DOI: 10.1016/j.fob.2015.06.007 PMCID: PMC4506957 PMID: 26199863
http://www.ncbi.nlm.nih.gov/pubmed/23745983
1. J Proteome Res. 2013 Jul 5;12(7):3277-87. doi: 10.1021/pr400127j. Epub 2013 Jun 19. Improved N(α)-acetylated peptide enrichment following dimethyl labeling and SCX. Chen SH(1), Chen CR, Chen SH, Li DT, Hsu JL. Author information: (1)Department of Biological Science and Technology, National Pingtung University of Science and Technology, Pingtung, Taiwan. Protein N-terminal acetylation is one of the most common modifications occurring co- and post-translationally on either eukaryote or prokaryote proteins. However, compared to other protein modifications, the physiological role of protein N-terminal acetylation is relatively unclear. To explore the biological functions of protein N-terminal acetylation, a robust and large-scale method for qualitative and quantitative analysis of this modification is required. Enrichment of N(α)-acetylated peptides or depletion of the free N-terminal and internal tryptic peptides prior to analysis by mass spectrometry are necessary based on current technologies. This study demonstrated a simple strong cation exchange (SCX) fractionation method to selectively enrich N(α)-acetylated tryptic peptides via dimethyl labeling without the need for tedious protective labeling and depleting procedures. This method was introduced for the comprehensive analysis of N-terminal acetylated proteins from HepG2 cells. Several hundred N-terminal acetylation sites were readily identified in a single SCX flow-through fraction. Moreover, the N(α)-acetylated peptides of some protein isoforms were simultaneously observed in the SCX flow-through fraction, which indicated that this approach can be utilized to discriminate protein isoforms with very similar full sequences but different N-terminal sequences, such as β-actin/γ-actin, ERK1/ERK2, α-centractin/β-centractin, and ADP/ATP translocase 2 and 3. Compared to other methods, this method is relatively simple and can be directly implemented in a two-dimensional separation (SCX-RP)-mass spectrometry scheme for quantitative N-terminal proteomics using stable-isotope dimethyl labeling. DOI: 10.1021/pr400127j PMID: 23745983 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/20627866
1. Mol Cell Proteomics. 2010 Oct;9(10):2327-33. doi: 10.1074/mcp.M110.001271. Epub 2010 Jul 13. A quantitative proteomics design for systematic identification of protease cleavage events. Impens F(1), Colaert N, Helsens K, Ghesquière B, Timmerman E, De Bock PJ, Chain BM, Vandekerckhove J, Gevaert K. Author information: (1)Department of Medical Protein Research, VIB, B-9000 Ghent, Belgium. We present here a novel proteomics design for systematic identification of protease cleavage events by quantitative N-terminal proteomics, circumventing the need for time-consuming manual validation. We bypass the singleton detection problem of protease-generated neo-N-terminal peptides by introducing differential isotopic proteome labeling such that these substrate reporter peptides are readily distinguished from all other N-terminal peptides. Our approach was validated using the canonical human caspase-3 protease and further applied to mouse cathepsin D and E substrate processing in a mouse dendritic cell proteome, identifying the largest set of protein protease substrates ever reported and gaining novel insight into substrate specificity differences of these cathepsins. DOI: 10.1074/mcp.M110.001271 PMCID: PMC2953924 PMID: 20627866 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/26557869
1. Evid Based Complement Alternat Med. 2015;2015:983139. doi: 10.1155/2015/983139. Epub 2015 Oct 19. Recent Advance in Applications of Proteomics Technologies on Traditional Chinese Medicine Research. Ji Q(1), Zhu F(2), Liu X(3), Li Q(3), Su SB(4). Author information: (1)Research Center for Traditional Chinese Medicine Complexity System, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China ; Department of Medical Oncology, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China. (2)Jiangsu Provincial Academy of Traditional Chinese Medicine, Nanjing 210028, China. (3)Department of Medical Oncology, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China. (4)Research Center for Traditional Chinese Medicine Complexity System, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China. Proteomics technology, a major component of system biology, has gained comprehensive attention in the area of medical diagnosis, drug development, and mechanism research. On the holistic and systemic theory, proteomics has a convergence with traditional Chinese medicine (TCM). In this review, we discussed the applications of proteomic technologies in diseases-TCM syndrome combination researches. We also introduced the proteomic studies on the in vivo and in vitro effects and underlying mechanisms of TCM treatments using Chinese herbal medicine (CHM), Chinese herbal formula (CHF), and acupuncture. Furthermore, the combined studies of proteomics with other "-omics" technologies in TCM were also discussed. In summary, this report presents an overview of the recent advances in the application of proteomic technologies in TCM studies and sheds a light on the future global and further research on TCM. DOI: 10.1155/2015/983139 PMCID: PMC4629032 PMID: 26557869
http://www.ncbi.nlm.nih.gov/pubmed/23137709
1. J Proteomics. 2013 Jan 14;78:558-77. doi: 10.1016/j.jprot.2012.10.023. Epub 2012 Nov 6. Meat science: From proteomics to integrated omics towards system biology. D'Alessandro A(1), Zolla L. Author information: (1)Department of Ecological and Biological Sciences, University of Tuscia, Largo dell'Università, snc, 01100 Viterbo, Italy. Since the main ultimate goal of farm animal raising is the production of proteins for human consumption, research tools to investigate proteins play a major role in farm animal and meat science. Indeed, proteomics has been applied to the field of farm animal science to monitor in vivo performances of livestock animals (growth performances, fertility, milk quality etc.), but also to further our understanding of the molecular processes at the basis of meat quality, which are largely dependent on the post mortem biochemistry of the muscle, often in a species-specific way. Post mortem alterations to the muscle proteome reflect the biological complexity of the process of "muscle to meat conversion," a process that, despite decades of advancements, is all but fully understood. This is mainly due to the enormous amounts of variables affecting meat tenderness per se, including biological factors, such as animal species, breed specific-characteristic, muscle under investigation. However, it is rapidly emerging that the tender meat phenotype is not only tied to genetics (livestock breeding selection), but also to extrinsic factors, such as the rearing environment, feeding conditions, physical activity, administration of hormonal growth promotants, pre-slaughter handling and stress, post mortem handling. From this intricate scenario, biochemical approaches and systems-wide integrated investigations (metabolomics, transcriptomics, interactomics, phosphoproteomics, mathematical modeling), which have emerged as complementary tools to proteomics, have helped establishing a few milestones in our understanding of the events leading from muscle to meat conversion. The growing integration of omics disciplines in the field of systems biology will soon contribute to take further steps forward. Copyright © 2012 Elsevier B.V. All rights reserved. DOI: 10.1016/j.jprot.2012.10.023 PMID: 23137709 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23369275
1. J Biosci Bioeng. 2013 Jun;115(6):579-89. doi: 10.1016/j.jbiosc.2012.12.007. Epub 2013 Jan 29. Current metabolomics: practical applications. Putri SP(1), Nakayama Y, Matsuda F, Uchikata T, Kobayashi S, Matsubara A, Fukusaki E. Author information: (1)Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan. The field of metabolomics continues to grow rapidly over the last decade and has been proven to be a powerful technology in predicting and explaining complex phenotypes in diverse biological systems. Metabolomics complements other omics, such as transcriptomics and proteomics and since it is a 'downstream' result of gene expression, changes in the metabolome is considered to best reflect the activities of the cell at a functional level. Thus far, metabolomics might be the sole technology capable of detecting complex, biologically essential changes. As one of the omics technology, metabolomics has exciting applications in varied fields, including medical science, synthetic biology, medicine, and predictive modeling of plant, animal and microbial systems. In addition, integrated applications with genomics, transcriptomics, and proteomics provide greater understanding of global system biology. In this review, we discuss recent applications of metabolomics in microbiology, plant, animal, food, and medical science. Copyright © 2013. Published by Elsevier B.V. DOI: 10.1016/j.jbiosc.2012.12.007 PMID: 23369275 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/24957891
1. Metabolites. 2013 Feb 4;3(1):72-100. doi: 10.3390/metabo3010072. Recent applications of metabolomics toward cyanobacteria. Schwarz D(1), Orf I(2), Kopka J(3), Hagemann M(4). Author information: (1)Institut Biowissenschaften, Pflanzenphysiologie, Universität Rostock, Albert-Einstein-Str. 3, D-18059 Rostock, Germany. doreen.schwarz@uni-rostock.de. (2)Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, 14476 Golm, Germany. orf@mpimp-golm.mpg.de. (3)Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, 14476 Golm, Germany. kopka@mpimp-golm.mpg.de. (4)Institut Biowissenschaften, Pflanzenphysiologie, Universität Rostock, Albert-Einstein-Str. 3, D-18059 Rostock, Germany. martin.hagemann@uni-rostock.de. Our knowledge on cyanobacterial molecular biology increased tremendously by the application of the "omics" techniques. Only recently, metabolomics was applied systematically to model cyanobacteria. Metabolomics, the quantitative estimation of ideally the complete set of cellular metabolites, is particularly well suited to mirror cellular metabolism and its flexibility under diverse conditions. Traditionally, small sets of metabolites are quantified in targeted metabolome approaches. The development of separation technologies coupled to mass-spectroscopy- or nuclear-magnetic-resonance-based identification of low molecular mass molecules presently allows the profiling of hundreds of metabolites of diverse chemical nature. Metabolome analysis was applied to characterize changes in the cyanobacterial primary metabolism under diverse environmental conditions or in defined mutants. The resulting lists of metabolites and their steady state concentrations in combination with transcriptomics can be used in system biology approaches. The application of stable isotopes in fluxomics, i.e. the quantitative estimation of carbon and nitrogen fluxes through the biochemical network, has only rarely been applied to cyanobacteria, but particularly this technique will allow the making of kinetic models of cyanobacterial systems. The further application of metabolomics in the concert of other "omics" technologies will not only broaden our knowledge, but will also certainly strengthen the base for the biotechnological application of cyanobacteria. DOI: 10.3390/metabo3010072 PMCID: PMC3901253 PMID: 24957891
http://www.ncbi.nlm.nih.gov/pubmed/11509578
1. J Biol Chem. 2001 Oct 19;276(42):39290-4. doi: 10.1074/jbc.M107026200. Epub 2001 Aug 16. mDia-interacting protein acts downstream of Rho-mDia and modifies Src activation and stress fiber formation. Satoh S(1), Tominaga T. Author information: (1)Department of Molecular and Cell Biology, Dokkyo University School of Medicine, Tochigi 321-0293, Japan. The formin homology protein mDia is a Rho GTPase effector protein that participates in stress fiber formation, cytokinesis, and transcriptional activation of the serum response factor. Although the function of another effector of Rho, Rho-associated kinase, is well established, relatively little is known about the functional mechanism and the downstream targets of mDia. Our recent report of a Rho-mDia-Src-tyrosine kinase pathway suggested an important role for mDia in cell adhesion turnover. We identified a new mDia-interacting protein which is expressed ubiquitously. The new protein mainly binds to the proline-rich region of mDia through its Src homology 3 domain and also binds to Grb2 through its proline-rich domain. The protein is localized at the cell periphery and membrane ruffles and co-localizes with mDia. Co-expression of vSrc and the mDia-interacting protein induces significant morphological changes at focal contacts and activation of vSrc. Furthermore, we found that the mDia-interacting protein plays an important role in stress fiber formation induced by active mDia1. Our results suggest that this new protein regulates actin polymerization and cell adhesion turnover in the downstream portion of the Rho-mDia pathway by interacting with Grb2 and Src. DOI: 10.1074/jbc.M107026200 PMID: 11509578 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/16361707
1. J Biol Chem. 2006 Feb 17;281(7):4300-7. doi: 10.1074/jbc.M510277200. Epub 2005 Dec 18. The basic region of the diaphanous-autoregulatory domain (DAD) is required for autoregulatory interactions with the diaphanous-related formin inhibitory domain. Wallar BJ(1), Stropich BN, Schoenherr JA, Holman HA, Kitchen SM, Alberts AS. Author information: (1)Chemistry Department and Cell and Molecular Biology Program, Grand Valley State University, Allendale, Michigan 49401-9403, USA. Mammalian diaphanous-related (mDia) formins act as Rho GTPase effectors during cytoskeletal remodeling. Rho binding to mDia amino-terminal GTPase-binding domains (GBDs) causes the adjacent Dia-inhibitory domain (DID) to release the carboxyl-terminal Dia-autoregulatory (DAD) domain that flanks the formin homology-2 (FH2) domain. The release of DAD allows the FH2 domain to then nucleate and elongate nonbranched actin filaments. DAD, initially discovered as a region of homology shared between a phylogenetically divergent set of formin proteins, is comprised of a core motif, MDXLLXL, and an adjacent region is comprised of numerous basic residues, typically RRKR in the mDia family. Here, we show that these specific amino acids within the basic region of DAD contribute to the binding of DID and therefore the maintenance of the mDia autoregulatory mechanism. In addition, expression of full-length versions of mDia2 containing amino acid substitutions in either the DAD core or basic regions causes profound changes in the F-actin architecture, including the formation of filopodia-like structures that rapidly elongate from the cell edge. These studies further refine our understanding of the molecular contribution of DAD to mDia control and the role of mDia2 in the assembly of membrane protrusions. DOI: 10.1074/jbc.M510277200 PMID: 16361707 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/10629846
1. Nihon Yakurigaku Zasshi. 1999 Oct;114 Suppl 1:1P-5P. doi: 10.1254/fpj.114.supplement_1. [Cellular functions & pharmacological manipulations of the small GTPase Rho & Rho effectors]. [Article in Japanese] Narumiya S(1). Author information: (1)Department of Pharmacology, Faculty of Medicine, Kyoto University, Japan. Rho is a member of the Ras-related family of small molecular weight GTP-binding proteins, and works as a molecular switch by shuttling between the GDP-bound inactive form and the GTP-bound active form. Cellular functions of Rho have been studied by two ways; one is to express or microinject constitutively active Rho mutants in cells to identify the active phenotype of Rho, and the other is to use botulinum C3 exoenzyme that specifically ADP-ribosylates and inactivates Rho in cells to find out phenotypes of Rho inactivation. These analyses have revealed that Rho is involved in cell to substratum adhesion and motility, cell contraction and cytokinesis through the reorganization of the actincytoskeleton and modulation of its activity. These actions of Rho are mediated by downstream Rho effectors. Several putative Rho effectors have been isolated on the basis of their selective binding to the GTP-bound form of Rho. Among them, the ROCK family of Rho-associated serine/threonine protein kinases inactivates cofilin and myosin phosphatase to induce stabilization of filamentous actin and increase in the actomyosin-based contractility. mDia binds profilin likely to promote actin polymerization. Thus, these effectors in combination are supposed to work in organization of various forms of the actin cytoskeleton. Furthermore, analyses using a ROCK specific inhibitor Y-27632 have suggested that the Rho-ROCK pathway works in contractions of vascular and bronchial smooth muscles under various pathological conditions and is involved in malignant cell transformation and tumor invasion and metastasis. DOI: 10.1254/fpj.114.supplement_1 PMID: 10629846 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23256674
1. Expert Rev Proteomics. 2012 Dec;9(6):635-48. doi: 10.1586/epr.12.61. Deciphering the single-cell omic: innovative application for translational medicine. Mannello F(1), Ligi D, Magnani M. Author information: (1)Department of Biomolecular Sciences, Section of Clinical Biochemistry, Unit of Cell Biology, University Carlo Bo, Via O Ubaldini 7, 61029 Urbino (PU), Italy. ferdinando.mannello@uniurb.it Traditional technologies to investigate system biology are limited by the detection of parameters resulting from the averages of large populations of cells, missing cells produced in small numbers, and attempting to uniform the heterogeneity. The advent of proteomics and genomics at a single-cell level has set the basis for an outstanding improvement in analytical technology and data acquisition. It has been well demonstrated that cellular heterogeneity is closely related to numerous stochastic transcriptional events leading to variations in patterns of expression among single genetically identical cells. The new-generation technology of single-cell analysis is able to better characterize a cell's population, identifying and differentiating outlier cells, in order to provide both a single-cell experiment and a corresponding bulk measurement, through the identification, quantification and characterization of all system biology aspects (genomics, transcriptomics, proteomics, metabolomics, degradomics and fluxomics). The movement of omics into single-cell analysis represents a significant and outstanding shift. DOI: 10.1586/epr.12.61 PMID: 23256674 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/22309662
1. Expert Rev Vaccines. 2012 Feb;11(2):133-44. doi: 10.1586/erv.11.177. Targeting EGF receptor variant III: tumor-specific peptide vaccination for malignant gliomas. Del Vecchio CA(1), Li G, Wong AJ. Author information: (1)Stanford University, 1201 Welch Road, MSLS P105, Stanford, CA 94305, USA. Glioblastoma multiforme (GBM) is the most common and deadly of the human brain cancers. The EGF receptor is often amplified in GBM and provides a potential therapeutic target. However, targeting the normal receptor is complicated by its nearly ubiquitous and high level of expression in certain tissues. A naturally occurring deletion mutant of the EGF receptor, EGFRvIII, is a constitutively active variant originally identified in a high percentage of brain cancer cases, and more importantly is rarely found in normal tissue. A peptide vaccine, rindopepimut (CDX-110, Celldex Therapeutics), is directed against the novel exon 1-8 junction produced by the EGFRvIII deletion, and it has shown high efficacy in preclinical models. Recent Phase II clinical trials in patients with newly diagnosed GBM have shown EGFRvIII-specific immune responses and significantly increased time to progression and overall survival in those receiving vaccine therapy, as compared with published results for standard of care. Rindopepimut therefore represents a very promising therapy for patients with GBM. DOI: 10.1586/erv.11.177 PMID: 22309662 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/21154166
1. Curr Opin Mol Ther. 2010 Dec;12(6):741-54. Rindopepimut, a 14-mer injectable peptide vaccine against EGFRvIII for the potential treatment of glioblastoma multiforme. Del Vecchio CA(1), Wong AJ. Author information: (1)Stanford University, 300 Pasteur Drive, Edwards R213, Stanford, CA 94305, USA. Celldex Therapeutics is developing rindopepimut (CDX-110), a 14-mer injectable peptide vaccine for the potential treatment of glioblastoma multiforme (GBM). Rindopepimut specifically targets a novel junctional epitope of the EGFR deletion mutant EGFRvIII, which is a constitutively active receptor that is expressed in approximately 60 to 70% of patients with GBM. EGFRvIII expression is correlated with worse prognosis and reduced overall survival. Importantly, EGFRvIII is not expressed in normal brain tissue, making it an excellent therapeutic target. Preclinical studies demonstrated lasting tumor regression and increased survival times, as well as efficient generation of EGFRvIII-specific humoral and cellular immune responses, in animals expressing EGFRvIII and vaccinated with rindopepimut. Phase I and II clinical trials in patients with GBM demonstrated significantly increased median time to progression and overall survival time in those vaccinated with rindopepimut compared with matched historical controls. Only limited side effects have been observed in patients. Given these results, rindopepimut is an extremely promising therapy for patients with GBM. Phase I and II clinical trials in patients with GBM were ongoing at the time of publication. In the future, it may be beneficial to combine rindopepimut with other treatment modalities to further prolong survival. PMID: 21154166 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/18159245
1. PLoS One. 2007 Dec 26;2(12):e1362. doi: 10.1371/journal.pone.0001362. Noncompaction of the ventricular myocardium is associated with a de novo mutation in the beta-myosin heavy chain gene. Budde BS(1), Binner P, Waldmüller S, Höhne W, Blankenfeldt W, Hassfeld S, Brömsen J, Dermintzoglou A, Wieczorek M, May E, Kirst E, Selignow C, Rackebrandt K, Müller M, Goody RS, Vosberg HP, Nürnberg P, Scheffold T. Author information: (1)Cologne Center for Genomics and Institute for Genetics, University of Cologne, Cologne, Germany. Noncompaction of the ventricular myocardium (NVM) is the morphological hallmark of a rare familial or sporadic unclassified heart disease of heterogeneous origin. NVM results presumably from a congenital developmental error and has been traced back to single point mutations in various genes. The objective of this study was to determine the underlying genetic defect in a large German family suffering from NVM. Twenty four family members were clinically assessed using advanced imaging techniques. For molecular characterization, a genome-wide linkage analysis was undertaken and the disease locus was mapped to chromosome 14ptel-14q12. Subsequently, two genes of the disease interval, MYH6 and MYH7 (encoding the alpha- and beta-myosin heavy chain, respectively) were sequenced, leading to the identification of a previously unknown de novo missense mutation, c.842G>C, in the gene MYH7. The mutation affects a highly conserved amino acid in the myosin subfragment-1 (R281T). In silico simulations suggest that the mutation R281T prevents the formation of a salt bridge between residues R281 and D325, thereby destabilizing the myosin head. The mutation was exclusively present in morphologically affected family members. A few members of the family displayed NVM in combination with other heart defects, such as dislocation of the tricuspid valve (Ebstein's anomaly, EA) and atrial septal defect (ASD). A high degree of clinical variability was observed, ranging from the absence of symptoms in childhood to cardiac death in the third decade of life. The data presented in this report provide first evidence that a mutation in a sarcomeric protein can cause noncompaction of the ventricular myocardium. DOI: 10.1371/journal.pone.0001362 PMCID: PMC2137931 PMID: 18159245 [Indexed for MEDLINE] Conflict of interest statement: Competing Interests: The authors have declared that no competing interests exist.
http://www.ncbi.nlm.nih.gov/pubmed/8389710
1. Eur Heart J. 1993 May;14(5):629-33. doi: 10.1093/eurheartj/14.5.629. Triiodothyronine therapy in open-heart surgery: from hope to disappointment. Teiger E(1), Menasché P, Mansier P, Chevalier B, Lajeunie E, Bloch G, Piwnica A. Author information: (1)Department of Cardiovascular Surgery, Hopital Lariboisière, Paris, France. A controversy persists as to whether cardiopulmonary bypass (CPB) decreases plasma levels of triiodothyronine (T3), thereby justifying peri-operative administration of T3 to improve haemodynamic recovery. To examine the effects of T3 therapy on post-CPB haemodynamics and to determine whether the potential inotropic effects of T3 are mediated by an increase in beta-adrenergic responsiveness, a prospective, randomized, double-blind, placebo-controlled study was performed in 20 patients undergoing cardiac surgery with CPB. T3 or placebo solution (10 patients in each group) was given intravenously at the time of aortic unclamping and 4, 8, 12 and 20 h thereafter. End points included (1) thyroid hormone levels measured by radioimmunoassay (2) standard haemodynamic parameters (3) the density of lymphocyte beta-adrenoceptors measured by a radioligand (125I-iodocyanopindolol) binding technique. Post-CPB values (cross clamp removal) of T3 (pg.ml-1) were not significantly decreased compared with pre-CPB values: 3.3 +/- 0.2 vs 3.1 +/- 0.2 in controls and 3.3 +/- 0.4 vs 3.7 +/- 0.6 in T3-treated patients, respectively. The haemodynamic parameters were no different between the two groups at any postoperative time point. Likewise, density and affinity of lymphocyte beta-adrenoceptors were not significantly different from pre-operative values in either group. Thus, there seems to be no sound justification for a routine use of T3 in patients undergoing open-heart procedures. DOI: 10.1093/eurheartj/14.5.629 PMID: 8389710 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/8594265
1. JAMA. 1996 Mar 6;275(9):687-92. Cardiovascular effects of intravenous triiodothyronine in patients undergoing coronary artery bypass graft surgery. A randomized, double-blind, placebo- controlled trial. Duke T3 study group. Bennett-Guerrero E(1), Jimenez JL, White WD, D'Amico EB, Baldwin BI, Schwinn DA. Author information: (1)Department of Anesthesiology, Duke University Medical Center, Durham, NC, USA. Comment in JAMA. 1996 Mar 6;275(9):723-4. JAMA. 1996 Jul 10;276(2):100-1. OBJECTIVE: To test the hypothesis that triiodothyronine (T(3)) administration improves hemodynamic variables and decreases inotropic drug requirements in cardiac surgery patients. DESIGN: Prospective, randomized, double-blind, placebo-controlled trial. SETTING: Tertiary care medical center. PATIENTS: A total of 211 patients undergoing coronary artery surgery at high risk for requiring inotropic drug support. INTERVENTION: At release of aortic cross-clamp, patients were randomized to an intravenous infusion of T(3) (0.8 microg/kg followed by 0.12 microg.kg(-1).h(-1) for 6 hours), dopamine (positive control, 5 microg.kg(-1).min(-1) for 6 hours) or placebo. MAIN OUTCOME MEASURES: Perioperative hemodynamic variables, inotropic support requirements, and serum T(3) concentrations. RESULTS: Mean+/-SEM free T(3) serum concentrations decreased significantly during cardiopulmonary bypass in all groups (from 0.0035+/-0.0001 nmol/L [0.23+/-0.01 ng/dL] to 0.001+/-0.0001 nmol/L [0.7+/- 0.00 ng/dL]; P=.001) and increased to 0.0133+/-0.0004 nmol/L [0.87+/-0.03 ng/dL] (twice normal range; P<.001) following initiation of intravenous T(3). Intravenous T(3) did not change hemodynamic variables or inotropic drug requirements; however, heart rate increased (P<.001), and a trend toward decreased use of inotropic agents was demonstrated in the dopamine group. CONCLUSIONS: Triiodothyronine administration prevents decreases in serum thyroid hormone concentrations associated with cardiopulmonary bypass. Intravenous T(3) does not have dramatic effects on hemodynamic variables in this setting as has been previously suggested. Although mild effects on myocardial performance may exist, we cannot recommend at this time the routine use of intravenous T(3) as an inotropic agent in patients undergoing coronary artery bypass graft surgery. PMID: 8594265 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23319591
1. J Biol Chem. 2013 Mar 22;288(12):8380-8390. doi: 10.1074/jbc.M112.359935. Epub 2013 Jan 14. Methylation of histone H3 on lysine 4 by the lysine methyltransferase SET1 protein is needed for normal clock gene expression. Raduwan H(1), Isola AL(1), Belden WJ(2). Author information: (1)Department of Biochemistry and Microbiology, Rutgers, The State University of New Jersey, New Brunswick, New Jersey 08901. (2)Department of Biochemistry and Microbiology, Rutgers, The State University of New Jersey, New Brunswick, New Jersey 08901. Electronic address: belden@aesop.rutgers.edu. The circadian oscillator controls time-of-day gene expression by a network of interconnected feedback loops and is reset by light. The requisite for chromatin regulation in eukaryotic transcription necessitates temporal regulation of histone-modifying and chromatin-remodeling enzymes for proper clock function. CHD1 is known to bind H3K4me3 in mammalian cells, and Neurospora CHD1 is required for proper regulation of the frequency (frq) gene. Based on this, we examined a strain lacking SET1 to determine the role of H3K4 methylation in clock- and light-mediated frq regulation. Expression of frq was altered in strains lacking set1 under both circadian- and light-regulated gene expression. There is a delay in the phasing of H3K4me3 relative to the peak in frq expression. White Collar 2 (WC-2) association with the frq promoter persists longer in Δset1, suggesting a more permissible chromatin state. Surprisingly, SET1 is required for DNA methylation in the frq promoter, indicating a dependence on H3K4me for DNA methylation. The data support a model where SET1 is needed for proper regulation by modulating chromatin at frq. DOI: 10.1074/jbc.M112.359935 PMCID: PMC3605655 PMID: 23319591 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/21285074
1. Eur Heart J. 2011 Jun;32(12):1446-56. doi: 10.1093/eurheartj/ehq508. Epub 2011 Jan 31. Left ventricular non-compaction revisited: a distinct phenotype with genetic heterogeneity? Oechslin E(1), Jenni R. Author information: (1)Toronto Congenital Cardiac Centre for Adults, Peter Munk Cardiac Centre, University Health Network/Toronto General Hospital, Toronto, Ontario, Canada. erwin.oechslin@uhn.on.ca Non-compaction of the left ventricular myocardium (LVNC) has gained increasing recognition during the last 25 years. There is a morphological trait of the myocardial structure with a spectrum from normal variants to the pathological phenotype of LVNC, which reflects the embryogenic structure of the human heart due to an arrest in the compaction process during the first trimester. It must be cautioned not to overdiagnose LVNC: the morphological spectrum of trabeculations, from normal variants to pathological trabeculations with the morphological feature of LVNC must be carefully considered. The classical triad of complications are heart failure, arrhythmias, including sudden cardiac death, and systemic embolic events. Non-compaction of the left ventricular myocardium can occur in isolation or in association with congenital heart defects (CHDs), genetic syndromes, and neuromuscular disorders among others. The clinical spectrum is wide and the outcome is more favourable than in previously described populations with a negative selection bias. Familial occurrence is frequent with autosomal dominant and X-linked transmissions. Different mutations in sarcomere protein genes were identified and there seems to be a shared molecular aetiology of different cardiomyopathic phenotypes, including LVNC, hypertrophic and dilated cardiomyopathies. Thus, genetic heterogeneity, with an overlap of different phenotypes, and the variability of hereditary patterns, raise the questions whether there is a morphological trait from dilated/hypertrophic cardiomyopathy to LVNC and what are the triggers and modifiers to develop either dilated, hypertrophic cardiomyopathy, or LVNC in patients with the same mutation. The variety in clinical presentation, the genetic heterogeneity, and the phenotype of the first transgenetic animal model of an LVNC-associated mutation question the hypothesis that LVNC be a distinct cardiomyopathy: it seems to be rather a distinct phenotype or phenotypic, morphological expression of different underlying diseases than a distinct cardiomyopathy. DOI: 10.1093/eurheartj/ehq508 PMID: 21285074 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/12857739
1. J Biol Chem. 2003 Oct 3;278(40):38902-12. doi: 10.1074/jbc.M306229200. Epub 2003 Jul 10. Activation of the Rac-binding partner FHOD1 induces actin stress fibers via a ROCK-dependent mechanism. Gasteier JE(1), Madrid R, Krautkrämer E, Schröder S, Muranyi W, Benichou S, Fackler OT. Author information: (1)Institute for Hygiene, Department of Virology, University of Heidelberg, 69120 Heidelberg, Germany. Diaphanous related formins (DRFs) are part of the formin protein family that control morphogenesis, embryonic differentiation, cytokinesis, and cell polarity. DRFs organize the cytoskeleton in eukaryotic cells via the interaction with specific members of the Rho family of small GTPases including Rho, Rac, and Cdc42. This is best understood for Rho, which transmits signals to the actin cytoskeleton through the cooperation of its DRF effector mDia with ROCK (Rho-associated kinase). Here, we show that a constitutive active form of the Rac-interacting DRF FHOD1 (formin homology 2 domain containing 1) associates with F-actin in NIH3T3 cells, resulting in the formation of thick actin fibers. Cytoskeletal changes induced by FHOD1 correlated with the induction of serum response element transcription and were mediated by formin homology domains 1 and 2 of FHOD1. FHOD1-induced effects required the activity of the Rho-ROCK cascade that is targeted at a level downstream of Rho by the DRF. However, when the functional interaction of FHOD1 with individual GTPases was addressed, Rac but not Rho or Cdc42 bound to FHOD1 in cells and induced its recruitment to actin filaments and lamellipodia/membrane ruffles. Furthermore, activated FHOD1 interfered with lamellipodia formation. These results indicate that FHOD1 acts as an effector of Rac in actin rearrangements and transcriptional regulation and may provide a link for the Rac-dependent activation of the Rho cascade. DOI: 10.1074/jbc.M306229200 PMID: 12857739 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/20083571
1. Cardiovasc Res. 2010 Jun 1;86(3):452-60. doi: 10.1093/cvr/cvq009. Epub 2010 Jan 18. Severe familial left ventricular non-compaction cardiomyopathy due to a novel troponin T (TNNT2) mutation. Luedde M(1), Ehlermann P, Weichenhan D, Will R, Zeller R, Rupp S, Müller A, Steen H, Ivandic BT, Ulmer HE, Kern M, Katus HA, Frey N. Author information: (1)Department of Internal Medicine III, University of Heidelberg, 69120 Heidelberg, Germany. AIMS: Left ventricular non-compaction (LVNC) is caused by mutations in multiple genes. It is still unclear whether LVNC is the primary determinant of cardiomyopathy or rather a secondary phenomenon with intrinsic cardiomyocyte dysfunction being the actual cause of the disease. Here, we describe a family with LVNC due to a novel missense mutation, pE96K, in the cardiac troponin T gene (TNNT2). METHODS AND RESULTS: The novel mutation was identified in the index patient and all affected relatives, but not in 430 healthy control individuals. Mutations in known LVNC-associated genes were excluded. To investigate the pathophysiological implications of the mutation, we generated transgenic mice expressing human wild-type cTNT (hcTNT) or a human troponin T harbouring the pE96K mutation (mut cTNT). Animals were characterized by echocardiography, histology, and gene expression analysis. Mut cTNT mice displayed an impaired left ventricular function and induction of marker genes of heart failure. Remarkably, left ventricular non-compaction was not observed. CONCLUSION: Familial co-segregation and the cardiomyopathy phenotype of mut cTNT mice strongly support a causal relationship of the pE96K mutation and disease in our index patient. In addition, our data suggest that a non-compaction phenotype is not required for the development of cardiomyopathy in this specific TNNT2 mutation leading to LVNC. DOI: 10.1093/cvr/cvq009 PMID: 20083571 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/7477166
1. N Engl J Med. 1995 Dec 7;333(23):1522-7. doi: 10.1056/NEJM199512073332302. Thyroid hormone treatment after coronary-artery bypass surgery. Klemperer JD(1), Klein I, Gomez M, Helm RE, Ojamaa K, Thomas SJ, Isom OW, Krieger K. Author information: (1)Department of Cardiothoracic Surgery, New York Hospital-Cornell University Medical College, New York 10021, USA. Comment in N Engl J Med. 1995 Dec 7;333(23):1562-3. doi: 10.1056/NEJM199512073332310. BACKGROUND: Thyroid hormone has many effects on the cardiovascular system. During and after cardiopulmonary bypass, serum triiodothyronine concentrations decline transiently, which may contribute to postoperative hemodynamic dysfunction. We investigated whether the perioperative administration of triiodothyronine (liothyronine sodium) enhances cardiovascular performance in high-risk patients undergoing coronary-artery bypass surgery. METHODS: We administered triiodothyronine or placebo to 142 patients with coronary artery disease and depressed left ventricular function. The hormone was administered as an intravenous bolus of 0.8 microgram per kilogram of body weight when the aortic cross-clamp was removed after the completion of bypass surgery and then as an infusion of 0.113 microgram per kilogram per hour for six hours. Clinical and hemodynamic responses were serially recorded, as was any need for inotropic or vasodilator drugs. RESULTS: The patients' preoperative serum triiodothyronine concentrations were normal (mean [+/- SD] value, 81 +/- 22 ng per deciliter [1.2 +/- 0.3 nmol per liter]), and they decreased by 40 percent (P < 0.001) 30 minutes after the onset of cardiopulmonary bypass. The concentrations in patients given intravenous triiodothyronine became supranormal and were significantly higher than those in patients given placebo (P < 0.001). However, the concentrations were once again similar in the two groups 24 hours after surgery. The mean postoperative cardiac index was higher in the triiodothyronine group (2.97 +/- 0.72 vs. 2.67 +/- 0.61 liters per minute per square meter of body-surface area, P = 0.007), and systemic vascular resistance was lower (1073 +/- 314 vs. 1235 +/- 387 dyn.sec.cm-5, P = 0.003). The two groups did not differ significantly in the incidence of arrhythmia or the need for therapy with inotropic and vasodilator drugs during the 24 hours after surgery, or in perioperative mortality and morbidity. CONCLUSIONS: Raising serum triiodothyronine concentrations in patients undergoing coronary-artery bypass surgery increases cardiac output and lowers systemic vascular resistance, but does not change outcome or alter the need for standard postoperative therapy. DOI: 10.1056/NEJM199512073332302 PMID: 7477166 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/22221061
1. Comb Chem High Throughput Screen. 2012 May 1;15(4):286-98. doi: 10.2174/138620712799361852. Integrative system biology strategies for disease biomarker discovery. Zhang H(1), Hu H, Deng C, Chun Y, Zhou S, Huang F, Zhou Q. Author information: (1)Medical School, Yangtze University, Jingzhou Hubei 434000, People's Republic of China. zhanghaiyuan1972@hotmail.com Biomarkers are currently widely used to diagnose diseases, monitor treatments, and evaluate potential drug candidates. Research of differential Omics accelerate the advancements of biomarkers' discovery. By extracting biological knowledge from the 'omics' through integration, integrative system biology creates predictive models of cells, organs, biochemical processes and complete organisms, in addition to identifying human disease biomarkers. Recent development in high-throughput methods enables analysis of genome, transcriptome, proteome, and metabolome at an unprecedented scale, thus contributing to the deluge of experimental data in numerous public databases. Several integrative system biology approaches have been developed and applied to the discovery of disease biomarkers from databases. In this review, we highlight several of these approaches and identify future steps in the context of the field of integrative system biology. DOI: 10.2174/138620712799361852 PMID: 22221061 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/18395448
1. Neuromuscul Disord. 2008 Apr;18(4):331-3. doi: 10.1016/j.nmd.2007.11.012. Left ventricular non-compaction in a patient with myotonic dystrophy type 2. Wahbi K(1), Meune C, Bassez G, Laforêt P, Vignaux O, Marmursztejn J, Bécane HM, Eymard B, Duboc D. Author information: (1)Department of Cardiology, Cochin Hospital, Assistance Publique Hôpitaux de Paris, René Descartes University, 27 rue du Fg St-Jacques, 75014 Paris, France. karim.wahbi@cch.aphp.fr Cardiac involvement is frequent in myotonic dystrophy type 2 (DM2) with dilated cardiomyopathy and severe arrhythmias having been reported. Left ventricular non-compaction is a cardiomyopathy often associated with neuromuscular disorders. We report the case of a 61-year-old man with DM2 treated for 5 years for a suspected dilated cardiomyopathy. Echocardiography showed left ventricular non-compaction typical pattern, with prominent apical trabeculations and intertrabecular spaces perfused from ventricular cavity. MRI confirmed the diagnosis. Physicians should be aware of the risk of severe cardiac complications in DM2 patients. Left ventricular non-compaction diagnosis is often overlooked. Neurological examination should be performed in all patients with left ventricular non-compaction. DOI: 10.1016/j.nmd.2007.11.012 PMID: 18395448 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/17101628
1. Europace. 2006 Dec;8(12):1064-7. doi: 10.1093/europace/eul125. Epub 2006 Nov 13. Isolated ventricular non-compaction: clinical study and genetic review. Markiewicz-Loskot G(1), Moric-Janiszewska E, Loskot M, Szydlowski L, Weglarz L, Hollek A. Author information: (1)Department of Pediatric Cardiology, Medical University of Silesia, Medyków 16, 40-752 Katowice, Poland. ejaniszewska@slam.katowice.pl Comment in Europace. 2007 May;9(5):333. doi: 10.1093/europace/eum012. Isolated non-compaction of the ventricular myocardium (INVM), sometimes referred to as 'spongy myocardium', is a congenital and exceedingly rare cardiomyopathy. Isolated ventricular non-compaction occurs in the absence of other structural heart diseases and, hypothetically, it is due to the arrest of myocardial morphogenesis. Isolated non-compaction of the ventricular myocardium may manifest itself from infancy to young adulthood with a high mortality rate. Both sexes are affected. In our study, we present a case of INVM (left and right ventricles) in a 3-year-old girl, diagnosed by two-dimensional echocardiography. The anomaly presented as a restrictive cardiomyopathy. The girl was admitted to our hospital with heart failure, when she was 10 months old. She was treated with dopamine, digoxin, furosemide, spironolactone, and acenocoumarol and her condition improved. Presently, the girl remains asymptomatic and for 3 years of follow-up, her development has been almost normal. We here describe the genetic background of this disorder (based on a literature review). DOI: 10.1093/europace/eul125 PMID: 17101628 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19478006
1. Bioinformatics. 2009 Jun 15;25(12):i321-9. doi: 10.1093/bioinformatics/btp230. DISCOVER: a feature-based discriminative method for motif search in complex genomes. Fu W(1), Ray P, Xing EP. Author information: (1)School of Computer Science, Carnegie Mellon University, Pittsburgh, PA 15213, USA. MOTIVATION: Identifying transcription factor binding sites (TFBSs) encoding complex regulatory signals in metazoan genomes remains a challenging problem in computational genomics. Due to degeneracy of nucleotide content among binding site instances or motifs, and intricate 'grammatical organization' of motifs within cis-regulatory modules (CRMs), extant pattern matching-based in silico motif search methods often suffer from impractically high false positive rates, especially in the context of analyzing large genomic datasets, and noisy position weight matrices which characterize binding sites. Here, we try to address this problem by using a framework to maximally utilize the information content of the genomic DNA in the region of query, taking cues from values of various biologically meaningful genetic and epigenetic factors in the query region such as clade-specific evolutionary parameters, presence/absence of nearby coding regions, etc. We present a new method for TFBS prediction in metazoan genomes that utilizes both the CRM architecture of sequences and a variety of features of individual motifs. Our proposed approach is based on a discriminative probabilistic model known as conditional random fields that explicitly optimizes the predictive probability of motif presence in large sequences, based on the joint effect of all such features. RESULTS: This model overcomes weaknesses in earlier methods based on less effective statistical formalisms that are sensitive to spurious signals in the data. We evaluate our method on both simulated CRMs and real Drosophila sequences in comparison with a wide spectrum of existing models, and outperform the state of the art by 22% in F1 score. AVAILABILITY AND IMPLEMENTATION: The code is publicly available at http://www.sailing.cs.cmu.edu/discover.html. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. DOI: 10.1093/bioinformatics/btp230 PMCID: PMC2687984 PMID: 19478006 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/16895927
1. Bioinformatics. 2006 Oct 15;22(20):2562-4. doi: 10.1093/bioinformatics/btl428. Epub 2006 Aug 7. HiRes--a tool for comprehensive assessment and interpretation of metabolomic data. Zhao Q(1), Stoyanova R, Du S, Sajda P, Brown TR. Author information: (1)Hatch Center for MR Research, Department of Radiology, Columbia University, New York, NY 10032 USA. qz2106@columbia.edu The increasing role of metabolomics in system biology is driving the development of tools for comprehensive analysis of high-resolution NMR spectral datasets. This task is quite challenging since unlike the datasets resulting from other 'omics', a substantial preprocessing of the data is needed to allow successful identification of spectral patterns associated with relevant biological variability. HiRes is a unique stand-alone software tool that combines standard NMR spectral processing functionalities with techniques for multi-spectral dataset analysis, such as principal component analysis and non-negative matrix factorization. In addition, HiRes contains extensive abilities for data cleansing, such as baseline correction, solvent peak suppression, removal of frequency shifts owing to experimental conditions as well as auxiliary information management. Integration of these components together with multivariate analytical procedures makes HiRes very capable of addressing the challenges for assessment and interpretation of large metabolomic datasets, greatly simplifying this otherwise lengthy and difficult process and assuring optimal information retrieval. AVAILABILITY: HiRes is freely available for research purposes at http://hatch.cpmc.columbia.edu/highresmrs.html DOI: 10.1093/bioinformatics/btl428 PMID: 16895927 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19438153
1. Rev Port Cardiol. 2009 Feb;28(2):185-94. Left ventricular non-compaction: a new mutation predisposing to reverse remodeling? [Article in English, Portuguese] Cortez-Dias N(1), Varela MG, Sargento L, Brito D, Almeida A, Cerqueira R, Lança V, Fernandes AR, Tavares P, Pereira RA, Fernandes A, Madeira H. Author information: (1)Serviço de Cardiologia, Hospital de Santa Maria, Lisboa, Portugal. Left ventricular non-compaction (LVNC) is a rare disorder of endomyocardial morphogenesis that results in multiple trabeculations and deep intertrabecular recesses filled with direct blood flow from the left ventricular cavity. LVNC is attracting increasing interest as a model for the study of cardiomyopathies, since it is a genetically heterogeneous disorder which varies greatly in clinical presentation and age of onset. The authors present the case of a young black male with progressive congestive heart failure of 2-3 years' evolution. The investigation, which included transthoracic echocardiography (contrast and 3D), transesophageal echocardiography and cardiac magnetic resonance imaging, showed LVNC and severe aortic regurgitation, with severe left ventricular systolic dysfunction. The family history was suggestive of genetically transmitted disease and genetic study of the TAZ gene at locus Xq28 identified the mutation p.Phe128Ser (c.383T>C), the first description of this mutation in a patient with LVNC. The patient underwent aortic valve replacement, with excellent clinical evolution, regression of left ventricular dimensions and global systolic functio Aortic regurgitation (not related to LVNC) was the determining factor in the clinical expression. However, the excellent reverse remodeling that occurred after surgery highlights the heterogeneity of myocardial behavior in LVNC patients. PMID: 19438153 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/21604106
1. Neth Heart J. 2013 Mar;21(3):113-7. doi: 10.1007/s12471-011-0141-1. Ebstein's anomaly may be caused by mutations in the sarcomere protein gene MYH7. van Engelen K(1), Postma AV, van de Meerakker JB, Roos-Hesselink JW, Helderman-van den Enden AT, Vliegen HW, Rahman T, Baars MJ, Sels JW, Bauer U, Pickardt T, Sperling SR, Moorman AF, Keavney B, Goodship J, Klaassen S, Mulder BJ. Author information: (1)Department of Cardiology, Academic Medical Center, Meibergdreef 9, 1105 AZ, Amsterdam, the Netherlands. Ebstein's anomaly is a rare congenital heart malformation characterised by adherence of the septal and posterior leaflets of the tricuspid valve to the underlying myocardium. Associated abnormalities of left ventricular morphology and function including left ventricular noncompaction (LVNC) have been observed. An association between Ebstein's anomaly with LVNC and mutations in the sarcomeric protein gene MYH7, encoding β-myosin heavy chain, has been shown by recent studies. This might represent a specific subtype of Ebstein's anomaly with a Mendelian inheritance pattern. In this review we discuss the association of MYH7 mutations with Ebstein's anomaly and LVNC and its implications for the clinical care for patients and their family members. DOI: 10.1007/s12471-011-0141-1 PMCID: PMC3578524 PMID: 21604106
http://www.ncbi.nlm.nih.gov/pubmed/15883375
1. Proc Natl Acad Sci U S A. 2005 May 17;102(20):7079-84. doi: 10.1073/pnas.0408743102. Epub 2005 May 9. De novo cis-regulatory module elicitation for eukaryotic genomes. Gupta M(1), Liu JS. Author information: (1)Department of Biostatistics, University of North Carolina, Chapel Hill, NC 27599, USA. gupta@bios.unc.edu Transcription regulation is controlled by coordinated binding of one or more transcription factors in the promoter regions of genes. In many species, especially higher eukaryotes, transcription factor binding sites tend to occur as homotypic or heterotypic clusters, also known as cis-regulatory modules. The number of sites and distances between the sites, however, vary greatly in a module. We propose a statistical model to describe the underlying cluster structure as well as individual motif conservation and develop a Monte Carlo motif screening strategy for predicting novel regulatory modules in upstream sequences of coregulated genes. We demonstrate the power of the method with examples ranging from bacterial to insect and human genomes. DOI: 10.1073/pnas.0408743102 PMCID: PMC1129096 PMID: 15883375 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/22619057
1. J Neurol. 2012 Jul;259(7):1494-6. doi: 10.1007/s00415-012-6543-1. Epub 2012 May 23. ZASPopathy with childhood-onset distal myopathy. Strach K(1), Reimann J, Thomas D, Naehle CP, Kress W, Kornblum C. Author information: (1)Department of Radiology, University of Bonn, Bonn, Germany. katharina.strach@med.ovgu.de We report on a German family presenting with a predominantly distal myopathy primarily affecting anterior compartments of lower legs in childhood. Proximal lower limb and hip girdle weakness developed later in early adulthood in the female index patient and likewise in her mother. Consecutive muscle biopsy findings were first attributed to a mild congenital myopathy and later on interpreted as neurogenic changes without clear signs of a myopathy. Molecular genetic analysis was performed because of the clinical impression of a distal myopathy combined with dominant inheritance. The heterozygous mutation c.349G>A (p.D117N) in the ZASP gene could be found. This mutation had been previously associated with an adult-onset, isolated, dilated left ventricular non-compaction cardiomyopathy (OMIM*605906.0007), which was not present in our patients. Our data show that this mutation can be associated with an isolated skeletal muscle phenotype. Second, mutation analysis of the ZASP gene is suggested for distal myopathies of any age, even in cases of uncharacteristic muscle biopsy findings on routine analysis. DOI: 10.1007/s00415-012-6543-1 PMID: 22619057 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/25268582
1. PLoS One. 2014 Sep 30;9(9):e108065. doi: 10.1371/journal.pone.0108065. eCollection 2014. CisMiner: genome-wide in-silico cis-regulatory module prediction by fuzzy itemset mining. Navarro C(1), Lopez FJ(2), Cano C(1), Garcia-Alcalde F(3), Blanco A(1). Author information: (1)Department of Computer Science and AI, University of Granada, Granada, Spain. (2)Andalusian Human Genome Sequencing Centre (CASEGH), Medical Genome Project (MGP), Sevilla, Spain. (3)Max Planck Institute for Infection Biology, Berlin, Germany. Eukaryotic gene control regions are known to be spread throughout non-coding DNA sequences which may appear distant from the gene promoter. Transcription factors are proteins that coordinately bind to these regions at transcription factor binding sites to regulate gene expression. Several tools allow to detect significant co-occurrences of closely located binding sites (cis-regulatory modules, CRMs). However, these tools present at least one of the following limitations: 1) scope limited to promoter or conserved regions of the genome; 2) do not allow to identify combinations involving more than two motifs; 3) require prior information about target motifs. In this work we present CisMiner, a novel methodology to detect putative CRMs by means of a fuzzy itemset mining approach able to operate at genome-wide scale. CisMiner allows to perform a blind search of CRMs without any prior information about target CRMs nor limitation in the number of motifs. CisMiner tackles the combinatorial complexity of genome-wide cis-regulatory module extraction using a natural representation of motif combinations as itemsets and applying the Top-Down Fuzzy Frequent- Pattern Tree algorithm to identify significant itemsets. Fuzzy technology allows CisMiner to better handle the imprecision and noise inherent to regulatory processes. Results obtained for a set of well-known binding sites in the S. cerevisiae genome show that our method yields highly reliable predictions. Furthermore, CisMiner was also applied to putative in-silico predicted transcription factor binding sites to identify significant combinations in S. cerevisiae and D. melanogaster, proving that our approach can be further applied genome-wide to more complex genomes. CisMiner is freely accesible at: http://genome2.ugr.es/cisminer. CisMiner can be queried for the results presented in this work and can also perform a customized cis-regulatory module prediction on a query set of transcription factor binding sites provided by the user. DOI: 10.1371/journal.pone.0108065 PMCID: PMC4182448 PMID: 25268582 [Indexed for MEDLINE] Conflict of interest statement: Competing Interests: The authors have declared that no competing interests exist.
http://www.ncbi.nlm.nih.gov/pubmed/21152003
1. PLoS Comput Biol. 2010 Dec 2;6(12):e1001020. doi: 10.1371/journal.pcbi.1001020. Assessing computational methods of cis-regulatory module prediction. Su J(1), Teichmann SA, Down TA. Author information: (1)MRC Laboratory of Molecular Biology, Cambridge, United Kingdom. Computational methods attempting to identify instances of cis-regulatory modules (CRMs) in the genome face a challenging problem of searching for potentially interacting transcription factor binding sites while knowledge of the specific interactions involved remains limited. Without a comprehensive comparison of their performance, the reliability and accuracy of these tools remains unclear. Faced with a large number of different tools that address this problem, we summarized and categorized them based on search strategy and input data requirements. Twelve representative methods were chosen and applied to predict CRMs from the Drosophila CRM database REDfly, and across the human ENCODE regions. Our results show that the optimal choice of method varies depending on species and composition of the sequences in question. When discriminating CRMs from non-coding regions, those methods considering evolutionary conservation have a stronger predictive power than methods designed to be run on a single genome. Different CRM representations and search strategies rely on different CRM properties, and different methods can complement one another. For example, some favour homotypical clusters of binding sites, while others perform best on short CRMs. Furthermore, most methods appear to be sensitive to the composition and structure of the genome to which they are applied. We analyze the principal features that distinguish the methods that performed well, identify weaknesses leading to poor performance, and provide a guide for users. We also propose key considerations for the development and evaluation of future CRM-prediction methods. DOI: 10.1371/journal.pcbi.1001020 PMCID: PMC2996316 PMID: 21152003 [Indexed for MEDLINE] Conflict of interest statement: The authors have declared that no competing interests exist.
http://www.ncbi.nlm.nih.gov/pubmed/12702576
1. Cancer Res. 2003 Apr 15;63(8):1871-5. SCCA2-like serpins mediate genetic predisposition to skin tumors. Gariboldi M(1), Peissel B, Fabbri A, Saran A, Zaffaroni D, Falvella FS, Spinola M, Tanuma J, Pazzaglia S, Mancuso MT, Maurichi A, Bartoli C, Cataltepe S, Silverman GA, Pilotti S, Hayashizaki Y, Okazaki Y, Dragani TA. Author information: (1)Department of Experimental Oncology, Istituto Nazionale Tumori 20133, Milan, Italy. Reasons for early onset skin cancer are poorly understood. Microarray analysis revealed overexpression of the Scca2 gene in the 12-O-tetradecanoylphorbol-13-acetate-treated skin of Car-S mice, or line phenotypically selected for high susceptibility to two-stage skin carcinogenesis, as compared with 12-O-tetradecanoylphorbol-13-acetate-treated skin of Car-R mice, which is resistant. A human skin squamous cell carcinoma cell line (NCI-H520) transfected with mouse Scca2 or a related gene, Scca2-rs1, both expressed in the skin, showed significantly increased tumor growth as compared with controls when injected in nude mice. Immunohistochemical analysis of samples from two independent series of Italian and Korean patients with squamous cell carcinoma of the skin indicated a significant association between SCCA2 protein expression and younger age at tumor onset. These findings provide evidence that SCCA2-like serpins mediate genetic predisposition to skin cancer in a mouse model and in humans. PMID: 12702576 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/14965369
1. Cancer Sci. 2004 Feb;95(2):176-80. doi: 10.1111/j.1349-7006.2004.tb03200.x. Proteasome inhibitor PS-341 induces growth arrest and apoptosis of non-small cell lung cancer cells via the JNK/c-Jun/AP-1 signaling. Yang Y(1), Ikezoe T, Saito T, Kobayashi M, Koeffler HP, Taguchi H. Author information: (1)Department of Internal Medicine, Kochi Medical School, Kochi 783-8505, Japan. Proteasome inhibitor PS-341 induces growth arrest and apoptosis of multiple myeloma (MM) cells via inactivation of NF-kappaB in vitro and has afforded some objective responses in individuals with relapsed, refractory MM. However, the activity of PS-341 against non-hematological malignancies remains to be fully elucidated. In this study, we found that PS-341 induced growth arrest and apoptosis of NCI-H520 and -H460 non-small cell lung cancer (NSCLC) cells in conjunction with markedly up-regulated levels of p21(waf1) and p53, and down-regulation of bcl-2 protein in these cells. Also, PS-341 caused phosphorylation of c-Jun NH(2)-terminal kinase (JNK) and c-Jun, and enhanced AP-1/DNA binding activities in these cells as measured by western blotting and enzyme-linked immunosorbent assay (ELISA), respectively. Interestingly, when the JNK/c-Jun/AP-1 signal pathway was disrupted by the JNK inhibitor SP600125, the ability of PS-341 to inhibit the growth of NSCLC cells and to up-regulate the levels of p21(waf1) in these cells was blunted, but the expression of p53 was sustained at a high level, suggesting that the JNK/c-Jun/AP-1 signal pathway might mediate the anti-lung cancer effects of PS-341, with p21(waf1) playing the central role. Thus, PS-341 might be useful for the treatment of individuals with NSCLC. DOI: 10.1111/j.1349-7006.2004.tb03200.x PMCID: PMC11160053 PMID: 14965369 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/14662268
1. J Am Coll Cardiol. 2003 Dec 3;42(11):2014-27. doi: 10.1016/j.jacc.2003.10.021. Mutations in Cypher/ZASP in patients with dilated cardiomyopathy and left ventricular non-compaction. Vatta M(1), Mohapatra B, Jimenez S, Sanchez X, Faulkner G, Perles Z, Sinagra G, Lin JH, Vu TM, Zhou Q, Bowles KR, Di Lenarda A, Schimmenti L, Fox M, Chrisco MA, Murphy RT, McKenna W, Elliott P, Bowles NE, Chen J, Valle G, Towbin JA. Author information: (1)Department of Pediatrics (Cardiology), Baylor College of Medicine, Houston, Texas, USA. Comment in J Am Coll Cardiol. 2004 Sep 1;44(5):1139; author reply 1139-40. doi: 10.1016/j.jacc.2004.06.007. OBJECTIVES: We evaluated the role of Cypher/ZASP in the pathogenesis of dilated cardiomyopathy (DCM) with or without isolated non-compaction of the left ventricular myocardium (INLVM). BACKGROUND: Dilated cardiomyopathy, characterized by left ventricular dilation and systolic dysfunction with signs of heart failure, is genetically transmitted in 30% to 40% of cases. Genetic heterogeneity has been identified with mutations in multiple cytoskeletal and sarcomeric genes causing the phenotype. In addition, INLVM with a hypertrophic dilated left ventricle, ventricular dysfunction, and deep trabeculations, is also inherited, and the genes identified to date differ from those causing DCM. Cypher/ZASP is a newly identified gene encoding a protein that is a component of the Z-line in both skeletal and cardiac muscle. METHODS: Diagnosis of DCM was performed by echocardiogram, electrocardiogram, and physical examination. In addition, levels of the muscular isoform of creatine kinase were measured to evaluate for skeletal muscle involvement. Cypher/ZASP was screened by denaturing high performance liquid chromatography (DHPLC) and direct deoxyribonucleic acid sequencing. RESULTS: We identified and screened 100 probands with left ventricular dysfunction. Five mutations in six probands (6% of cases) were identified in patients with familial or sporadic DCM or INLVM. In vitro studies showed cytoskeleton disarray in cells transfected with mutated Cypher/ZASP. CONCLUSIONS: These data suggest that mutated Cypher/ZASP can cause DCM and INLVM and identify a mechanistic basis. DOI: 10.1016/j.jacc.2003.10.021 PMID: 14662268 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/15750352
1. Mol Cells. 2005 Feb 28;19(1):143-8. Combined effect of heptaplatin and ionizing radiation on human squamous carcinoma cell lines. Ryu MR(1), Paik SY, Chung SM. Author information: (1)Department of Radiation Oncology, College of Medicine, The Catholic University of Korea, Seoul 137-701, Korea. Heptaplatin, cis-malonato [(4R,5R)-4,5-bis (amino-methyl)-2-isopropyl-1,3-dioxolane] platinum(II) (SKI-2053R, Sunpla) is a new platinum derivative with anti-tumor activity comparable to cisplatin on various cancer cell lines. Preclinical studies suggest that it is less nephrotoxic than cisplatin. This study was undertaken to examine the combined effect of heptaplatin and ionizing radiation on two established human squamous carcinoma cell lines (NCI-H520, SQ20B). The cytotoxic activity of heptaplatin was concentration-dependent in both cell lines. When low dose heptaplatin was combined with high dose ionizing radiation, there was an additive cytotoxic effect on NCI-H520 cells (P < 0.05), while a moderate dose of heptaplatin and a low dose of ionizing radiation had an additive cytotoxic effect on the growth of SQ20B cells (P < 0.05). FACS analysis and DAPI staining showed that their additive cytotoxic effects were correlated with the induction of apoptosis. Further studies are warranted using heptaplatin and ionizing radiation in squamous cell carcinoma as a substitute for cisplatin. PMID: 15750352 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23800289
1. Orphanet J Rare Dis. 2013 Jun 21;8:91. doi: 10.1186/1750-1172-8-91. Digenic mutational inheritance of the integrin alpha 7 and the myosin heavy chain 7B genes causes congenital myopathy with left ventricular non-compact cardiomyopathy. Esposito T(1), Sampaolo S, Limongelli G, Varone A, Formicola D, Diodato D, Farina O, Napolitano F, Pacileo G, Gianfrancesco F, Di Iorio G. Author information: (1)Institute of Genetics and Biophysics, National Research Council of Italy, Naples, Italy. teresa.esposito@igb.cnr.it Comment in Orphanet J Rare Dis. 2013 Nov 20;8:183. doi: 10.1186/1750-1172-8-183. BACKGROUND: We report an Italian family in which the proband showed a severe phenotype characterized by the association of congenital fiber type disproportion (CFTD) with a left ventricular non-compaction cardiomyopathy (LVNC). This study was focused on the identification of the responsible gene/s. METHODS AND RESULTS: Using the whole-exome sequencing approach, we identified the proband homozygous missense mutations in two genes, the myosin heavy chain 7B (MYH7B) and the integrin alpha 7 (ITGA7). Both genes are expressed in heart and muscle tissues, and both mutations were predicted to be deleterious and were not found in the healthy population.The R890C mutation in the MYH7B gene segregated with the LVNC phenotype in the examined family. It was also found in one unrelated patient affected by LVNC, confirming a causative role in cardiomyopathy.The E882K mutation in the ITGA7 gene, a key component of the basal lamina of muscle fibers, was found only in the proband, suggesting a role in CFTD. CONCLUSIONS: This study identifies two novel disease genes. Mutation in MYH7B causes a classical LVNC phenotype, whereas mutation in ITGA7 causes CFTD. Both phenotypes represent alterations of skeletal and cardiac muscle maturation and are usually not severe. The severe phenotype of the proband is most likely due to a synergic effect of these two mutations.This study provides new insights into the genetics underlying Mendelian traits and demonstrates a role for digenic inheritance in complex phenotypes. DOI: 10.1186/1750-1172-8-91 PMCID: PMC3695851 PMID: 23800289 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19467449
1. Trends Cardiovasc Med. 2009 Jan;19(1):17-21. doi: 10.1016/j.tcm.2009.03.003. Sarcomere mutations in cardiogenesis and ventricular noncompaction. McNally E(1), Dellefave L. Author information: (1)Department of Medicine, Section of Cardiology, The University of Chicago, Chicago, IL 60637, USA. emcnally@uchicago.edu Ventricular noncompaction is a form of cardiomyopathy where increased trabeculation is present frequently affecting the left ventricle and resembling an embryonic state of heart development. Clinically, left ventricular noncompaction may manifest as congestive heart failure, arrhythmias, and/or thromboembolic events. There are multiple genes linked to noncompaction, but recently, sarcomere gene mutations were found in both familial and sporadic cases of noncompaction. The association of noncompaction with sarcomere mutations supports the classification of ventricular noncompaction as cardiomyopathy and raises interesting questions regarding the continuum of hypertrophic cardiomyopathy, dilated cardiomyopathy, and noncompaction. The mutational spectrum of sarcomere genes in these disorders highlights the importance of the MYH7 gene encoding beta-myosin heavy chain and ACTC1 encoding the cardiac actin gene. Intriguingly, these mutations also share a low but definitive incidence of congenital heart malformations including septal defects. These human genetic findings support that normal myocardial and sarcomere function are required for proper compaction and septation and that these mutations also portend a high risk of developing heart failure in later life. DOI: 10.1016/j.tcm.2009.03.003 PMID: 19467449 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/20142034
1. Chem Biol. 2010 Jan 29;17(1):6-8. doi: 10.1016/j.chembiol.2010.01.002. Mechanism-based neddylation inhibitor. Petroski MD(1). Author information: (1)Signal Transduction Program, NCI Cancer Center, Burnham Institute for Medical Research, 10901 North Torrey Pines Road, La Jolla, CA 92037, USA. petroski@burnham.org Brownell et al. (2010) elucidate the mechanism of action of MLN4924, a NEDD8-activating enzyme inhibitor. MLN4924 requires the activity of the enzyme to generate a NEDD8-adenylate analog that potently and selectively shuts down this posttranslational modification system. Copyright (c) 2010 Elsevier Ltd. All rights reserved. DOI: 10.1016/j.chembiol.2010.01.002 PMID: 20142034
http://www.ncbi.nlm.nih.gov/pubmed/20542340
1. Int J Cardiol. 2010 Nov 19;145(2):405-407. doi: 10.1016/j.ijcard.2010.04.032. Epub 2010 Jun 14. The R820W mutation in the MYBPC3 gene, associated with hypertrophic cardiomyopathy in cats, causes hypertrophic cardiomyopathy and left ventricular non-compaction in humans. Ripoll Vera T(1), Monserrat Iglesias L(2), Hermida Prieto M(2), Ortiz M(2), Rodriguez Garcia I(2), Govea Callizo N(3), Gómez Navarro C(4), Rosell Andreo J(3), Gámez Martínez JM(5), Pons Lladó G(6), Cremer Luengos D(5), Torres Marqués J(5). Author information: (1)Cardiology Department, Hospital Son Llatzer, Palma de Mallorca, Spain. Electronic address: tripoll@hsll.es. (2)Instituto de Investigación Biomédica de A Coruña, Spain. (3)Genetics Unit, Hospital Son Dureta, Palma de Mallorca, Spain. (4)Cardiology Department, Hospital de Torrecárdenas, Almería, Spain. (5)Cardiology Department, Hospital Son Llatzer, Palma de Mallorca, Spain. (6)Cardiology Department, Clínica Palmaplanas, Palma de Mallorca, Spain. BACKGROUND: The R820W mutation in the MYBPC3 gene has been associated with the development of hypertrophic cardiomyopathy (HCM) in rag-doll cats, but had not been described in humans. AIMS: To describe the phenotype associated with the R820W mutation identified in a human family. METHODS: The R820W was identified by direct sequencing of the MYBPC3 gene in a 47 year old woman with HCM and left ventricular non-compaction (LVNC). Clinical and genetic studies of the R820W mutation were performed in her family. RESULTS: The index patient was homozygous for the mutation and had no additional mutations in the main sarcomeric genes (MYH7, TNNT2, TNNI3, and TPM1). She had HCM with LVNC and normal systolic function. One brother had died suddenly at age 43 years. Another brother diagnosed of LVNC with severe systolic dysfunction and a cardiac arrest was also homozygous for the mutation. One heterozygous 31 year old sister, and three heterozygous sons of the index (ages 14, 20 and 23 years old) were clinically unaffected. The father of the index was apparently healthy and her mother had atrial fibrillation and an electrocardiographic diagnosis of left ventricular hypertrophy at age 86 years. CONCLUSION: The R820W mutation in the MYBPC3 gene, previously associated with HCM in rag-doll cats, causes both HCM and LVNC in homozygous human carriers, with mild or null clinical expression in heterozygous carriers. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved. DOI: 10.1016/j.ijcard.2010.04.032 PMID: 20542340 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/25388161
1. Clin Cancer Res. 2015 Jan 15;21(2):439-47. doi: 10.1158/1078-0432.CCR-14-1960. Epub 2014 Nov 11. The NEDD8-activating enzyme inhibitor MLN4924 disrupts nucleotide metabolism and augments the efficacy of cytarabine. Nawrocki ST(1), Kelly KR(2), Smith PG(3), Keaton M(4), Carraway H(5), Sekeres MA(5), Maciejewski JP(5), Carew JS(6). Author information: (1)Division of Hematology/Oncology, CTRC, The University of Texas Health Science Center, San Antonio, Texas. (2)USC Norris Comprehensive Cancer Center, Pasadena, California. (3)Millennium Pharmaceuticals, Cambridge, Massachusetts. H3 Biomedicine, Cambridge, Massachusetts. (4)Metabolon, Inc., Durham, North Carolina. Duke University, Durham, North Carolina. (5)Taussig Cancer Institute, Cleveland Clinic, Cleveland, Ohio. (6)Taussig Cancer Institute, Cleveland Clinic, Cleveland, Ohio. carewj@ccf.org. PURPOSE: New therapies are urgently needed for patients with acute myelogenous leukemia (AML). The novel NEDDylation inhibitor MLN4924 (pevonedistat) has demonstrated significant preclinical antileukemic activity and preliminary efficacy in patients with AML in a phase I trial. On the basis of its antimyeloid and DNA-damaging properties, we investigated the ability of MLN4924 to augment conventional cytarabine (ara-C) therapy. EXPERIMENTAL DESIGN: The effects of MLN4924/ara-C on viability, clonogenic survival, apoptosis, DNA damage, and relevant pharmacodynamic targets were determined. The efficacy and pharmacodynamics of MLN4924/ara-C were assessed in an AML xenograft model. RESULTS: Cotreatment of AML cell lines and primary patient specimens with MLN4924 and ara-C led to diminished clonogenic survival, increased apoptosis, and synergistic levels of DNA damage. RNAi demonstrated that stabilization of CDT-1, an event previously shown to mediate the DNA-damaging effects of MLN4924, was not a key regulator of sensitivity to the MLN4924/ara-C combination. Global metabolic profiling revealed that MLN4924 disrupts nucleotide metabolism and depletes intracellular nucleotide pools in AML cells. Subsequent experiments showed that MLN4924 promoted increased incorporation of ara-C into the DNA of AML cells. This effect as well as the therapeutic benefit of the MLN4924/ara-C combination was antagonized by supplementation with the nucleotide building block ribose. Coadministration of MLN4924 and ara-C to mice bearing FLT3-ITD(+) AML xenografts stably inhibited disease progression and increased DNA damage in vivo. CONCLUSIONS: Our findings provide strong rationale for clinical investigation of the MLN4924/ara-C combination and establish a new link between therapeutic inhibition of NEDDylation and alterations in nucleotide metabolism. Clin Cancer Res; 21(2); 439-47. ©2014 AACR. ©2014 American Association for Cancer Research. DOI: 10.1158/1078-0432.CCR-14-1960 PMCID: PMC4297545 PMID: 25388161 [Indexed for MEDLINE] Conflict of interest statement: Disclosure of conflicts of interest: PGS is a former employee of Millennium Pharmaceuticals, Inc.
http://www.ncbi.nlm.nih.gov/pubmed/23147248
1. Biochim Biophys Acta. 2013 Apr;1833(4):833-9. doi: 10.1016/j.bbamcr.2012.11.003. Epub 2012 Nov 9. A novel alpha-tropomyosin mutation associates with dilated and non-compaction cardiomyopathy and diminishes actin binding. van de Meerakker JB(1), Christiaans I, Barnett P, Lekanne Deprez RH, Ilgun A, Mook OR, Mannens MM, Lam J, Wilde AA, Moorman AF, Postma AV. Author information: (1)Department of Anatomy, Embryology & Physiology, Academic Medical Center, Amsterdam, The Netherlands. BACKGROUND: Dilated cardiomyopathy (DCM) is characterized by idiopathic dilatation and systolic contractile dysfunction of the ventricle(s) leading to an impaired systolic function. The origin of DCM is heterogeneous, but genetic transmission of the disease accounts for up to 50% of the cases. Mutations in alpha-tropomyosin (TPM1), a thin filament protein involved in structural and regulatory roles in muscle cells, are associated with hypertrophic cardiomyopathy (HCM) and very rarely with DCM. METHODS AND RESULTS: Here we present a large four-generation family in which DCM is inherited as an autosomal dominant trait. Six family members have a cardiomyopathy with the age of diagnosis ranging from 5 months to 52 years. The youngest affected was diagnosed with dilated and non-compaction cardiomyopathy (NCCM) and died at the age of five. Three additional children died young of suspected heart problems. We mapped the phenotype to chromosome 15 and subsequently identified a missense mutation in TPM1, resulting in a p.D84N amino acid substitution. In addition we sequenced 23 HCM/DCM genes using next generation sequencing. The TPM1 p.D84N was the only mutation identified. The mutation co-segregates with all clinically affected family members and significantly weakens the binding of tropomyosin to actin by 25%. CONCLUSIONS: We show that a mutation in TPM1 is associated with DCM and a lethal, early onset form of NCCM, probably as a result of diminished actin binding caused by weakened charge-charge interactions. Consequently, the screening of TPM1 in patients and families with DCM and/or (severe, early onset forms of) NCCM is warranted. This article is part of a Special Issue entitled: Cardiomyocyte Biology: Cardiac Pathways of Differentiation, Metabolism and Contraction. Copyright © 2012 Elsevier B.V. All rights reserved. DOI: 10.1016/j.bbamcr.2012.11.003 PMID: 23147248 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/23715514
1. Oncotarget. 2013 May;4(5):739-50. doi: 10.18632/oncotarget.1034. Musashi1 as a potential therapeutic target and diagnostic marker for lung cancer. Wang XY(1), Yu H, Linnoila RI, Li L, Li D, Mo B, Okano H, Penalva LO, Glazer RI. Author information: (1)Cell and Cancer Biology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. Lung cancer remains one of the leading causes of cancer-related deaths worldwide with a 5-year survival rate of less than 20%. One approach to improving survival is the identification of biomarkers to detect early stage disease. In this study, we investigated the potential of the stem cell and progenitor cell marker, Musashi1 (Msi1), as a diagnostic marker and potential therapeutic target for lung cancer. Functional studies in A549 bronchioalveolar carcinoma and NCI-H520 squamous cell carcinoma cells revealed that Msi1 was enriched in spheroid cultures of tumor cells and in the CD133+ cell population. Downregulation of Msi1 by lentivirus-mediated expression of an Msi1 shRNA reduced spheroid colony proliferation. Growth inhibition was associated with reduced nuclear localization of β-catenin and inhibition of the processing of intracellular Notch. In primary lung cancer, Msi1 protein expression was elevated in 86% of 202 tissue microarray specimens, and Msi1 mRNA was increased in 80% of 118 bronchoscopic biopsies, including metastatic disease, but was rarely detected in adjacent normal lung tissue and in non-malignant diseased tissue. Msi1 was expressed in a diffuse pattern in most tumor subtypes, except in squamous cell carcinomas, where it appeared in a focal pattern in 50% of specimens. Thus, Msi1 is a sensitive and specific diagnostic marker for all lung cancer subtypes. DOI: 10.18632/oncotarget.1034 PMCID: PMC3742834 PMID: 23715514 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/11830536
1. Cancer Res. 2002 Feb 1;62(3):801-8. The CLN3 gene is a novel molecular target for cancer drug discovery. Rylova SN(1), Amalfitano A, Persaud-Sawin DA, Guo WX, Chang J, Jansen PJ, Proia AD, Boustany RM. Author information: (1)Department of Pediatrics, Duke University Medical Center, Durham, North Carolina 27710, USA. Juvenile Batten disease is a neurodegenerative disease caused by accelerated apoptotic death of photoreceptors and neurons attributable to defects in the CLN3 gene. CLN3 is antiapoptotic when overexpressed in NT2 neuronal precursor cells. CLN3 negatively modulates endogenous ceramide levels in NT2 cells and acts upstream of ceramide generation. Because defects in regulation of apoptosis are involved in the development of cancer, we evaluated the expression of CLN3 on both mRNA and protein levels in a variety of cancer cell lines and solid colon cancer tissue. We also observed the effect of the blocking of CLN3 protein expression on cancer cell growth, survival, ceramide production, and apoptosis by using an adenovirus-bearing antisense CLN3 construct. We show that CLN3 mRNA and protein are overexpressed in glioblastoma (U-373G and T98g), neuroblastoma (IMR-32 and SK-N-MC), prostate (Du145, PC-3, and LNCaP), ovarian (SK-OV-3, SW626, and PA-1), breast (BT-20, BT-549, and BT-474), and colon (SW1116, SW480, and HCT 116) cancer cell lines but not in pancreatic (CAPAN and As-PC-1) or lung (A-549 and NCI-H520) cancer cell lines. CLN3 is also up-regulated in mouse melanoma and breast carcinoma cancer cell lines. We found CLN3 expression is 22-330% higher than in corresponding normal colon control tissue in 8 of 10 solid colon tumors. An adenovirus-expressing antisense CLN3 (Ad-AS-CLN3) blocks CLN3 protein expression in DU-145, BT-20, SW1116, and T98g cancer cell lines as seen by Western blot. Blocking of CLN3 expression using Ad-AS-CLN3 inhibits growth and viability of cancer cells. It also causes elevation in endogenous ceramide production through de novo ceramide synthesis and results in increased apoptosis as shown by propidium iodide and JC-1 staining. This suggests that Ad-AS-CLN3 may be an option for therapy in some cancers. More importantly these results suggest that CLN3 is a novel molecular target for cancer drug discovery. PMID: 11830536 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/7482779
1. Trends Genet. 1995 Jul;11(7):283-90. doi: 10.1016/s0168-9525(00)89076-9. Dinucleotide relative abundance extremes: a genomic signature. Karlin S(1), Burge C. Author information: (1)Department of Mathematics, Stanford University, CA 94305-2125, USA. Early biochemical experiments established that the set of dinucleotide odds ratios or 'general design' is a remarkably stable property of the DNA of an organism, which is essentially the same in protein-coding DNA, bulk genomic DNA, and in different renaturation rate and density gradient fractions of genomic DNA in many organisms. Analysis of currently available genomic sequence data has extended these earlier results, showing that the general designs of disjoint samples of a genome are substantially more similar to each other than to those of sequences from other organisms and that closely related organisms have similar general designs. From this perspective, the set of dinucleotide odds ratio (relative abundance) values constitute a signature of each DNA genome, which can discriminate between sequences from different organisms. Dinucleotide-odds ratio values appear to reflect not only the chemistry of dinucleotide stacking energies and base-step conformational preferences, but also the species-specific properties of DNA modification, replication and repair mechanisms. DOI: 10.1016/s0168-9525(00)89076-9 PMID: 7482779 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/19968287
1. J Med Chem. 2010 Jan 14;53(1):139-46. doi: 10.1021/jm900803q. In vivo positron emission tomography (PET) imaging of mesenchymal-epithelial transition (MET) receptor. Wu C(1), Tang Z, Fan W, Zhu W, Wang C, Somoza E, Owino N, Li R, Ma PC, Wang Y. Author information: (1)Division of Radiopharmaceutical Science, Case Center for Imaging Research, Department of Radiology, Case Western Reserve University, Cleveland, Ohio 44106, USA. We report the radiosynthesis and evaluation of 3-[3,5-dimethyl-4-(4-[11C]methylpiperazinecarbonyl)-1H-pyrrol-2-ylmethylene]-2-oxo-2,3-dihydro-1H-indole-5-sulfonic acid (3-chlorophenyl)methylamide, termed [11C]SU11274 ([11C]14) for in vivo imaging of mesenchymal-epithelial transition (MET) receptor by positron emission tomography (PET). Following the synthesis of the precursor (13) that was achieved in 10 steps with a total yield of 9.7%, [11C]14 was obtained through radiomethylation in a range of 5-10% radiochemical yield and over 95% radiochemical purity. For in vivo PET studies, two human lung cancer xenograft models were established using MET-positive NCI-H1975 and MET-negative NCI-H520 cell lines. Quantitative [11C]14-PET studies showed that the tumor uptake of [11C]14 in the NCI-H1975 xenografts was significantly higher than that in the NCI-H520 xenografts, which is consistent with their corresponding immunohistochemical tissue staining patterns of MET receptors from the same animals. These studies demonstrated that [11C]14-PET is an appropriate imaging marker for quantification of MET receptor in vivo, which can facilitate efficacy evaluation in the clinical development of MET-targeted cancer therapeutics. DOI: 10.1021/jm900803q PMID: 19968287 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/17947214
1. Eur Heart J. 2007 Nov;28(22):2732-7. doi: 10.1093/eurheartj/ehm429. Epub 2007 Oct 17. Cardiac beta-myosin heavy chain defects in two families with non-compaction cardiomyopathy: linking non-compaction to hypertrophic, restrictive, and dilated cardiomyopathies. Hoedemaekers YM(1), Caliskan K, Majoor-Krakauer D, van de Laar I, Michels M, Witsenburg M, ten Cate FJ, Simoons ML, Dooijes D. Author information: (1)Department of Clinical Genetics, Erasmus Medical Centre, Dr Molewaterplein 50, 3015GE Rotterdam, The Netherlands. Comment in Eur Heart J. 2008 Apr;29(7):949-50; author reply 950-1. doi: 10.1093/eurheartj/ehn029. Cardiomyopathies are classified according to distinct morphological characteristics. They occur relatively frequent and are an important cause of mortality and morbidity. Isolated ventricular non-compaction or non-compaction cardiomyopathy (NCCM) is characterized by an excessively thickened endocardial layer with deep intertrabecular recesses, reminiscent of the myocardium during early embryogenesis. Aims Autosomal-dominant as well as X-linked inheritance for NCCM has been described and several loci have been associated with the disease. Nevertheless, a major genetic cause for familial NCCM remains to be identified. Methods and Results We describe, in two separate autosomal-dominant NCCM families, the identification of mutations in the sarcomeric cardiac beta-myosin heavy chain gene (MYH7), known to be associated with hypertrophic cardiomyopathy (HCM), restricted cardiomyopathy (RCM), and dilated cardiomyopathy (DCM). Conclusion These results confirm the genetic heterogeneity of NCCM and suggest that the molecular classification of cardiomyopathies includes an MYH7-associated spectrum of NCCM with HCM, RCM, and DCM. DOI: 10.1093/eurheartj/ehm429 PMID: 17947214 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/24129315
1. Mol Cell Proteomics. 2014 Jan;13(1):372-87. doi: 10.1074/mcp.O113.027870. Epub 2013 Oct 15. Immunoaffinity enrichment and mass spectrometry analysis of protein methylation. Guo A(1), Gu H, Zhou J, Mulhern D, Wang Y, Lee KA, Yang V, Aguiar M, Kornhauser J, Jia X, Ren J, Beausoleil SA, Silva JC, Vemulapalli V, Bedford MT, Comb MJ. Author information: (1)Cell Signaling Technology Inc., 3 Trask Lane, Danvers, Massachusetts 01923; Protein methylation is a common posttranslational modification that mostly occurs on arginine and lysine residues. Arginine methylation has been reported to regulate RNA processing, gene transcription, DNA damage repair, protein translocation, and signal transduction. Lysine methylation is best known to regulate histone function and is involved in epigenetic regulation of gene transcription. To better study protein methylation, we have developed highly specific antibodies against monomethyl arginine; asymmetric dimethyl arginine; and monomethyl, dimethyl, and trimethyl lysine motifs. These antibodies were used to perform immunoaffinity purification of methyl peptides followed by LC-MS/MS analysis to identify and quantify arginine and lysine methylation sites in several model studies. Overall, we identified over 1000 arginine methylation sites in human cell line and mouse tissues, and ∼160 lysine methylation sites in human cell line HCT116. The number of methylation sites identified in this study exceeds those found in the literature to date. Detailed analysis of arginine-methylated proteins observed in mouse brain compared with those found in mouse embryo shows a tissue-specific distribution of arginine methylation, and extends the types of proteins that are known to be arginine methylated to include many new protein types. Many arginine-methylated proteins that we identified from the brain, including receptors, ion channels, transporters, and vesicle proteins, are involved in synaptic transmission, whereas the most abundant methylated proteins identified from mouse embryo are transcriptional regulators and RNA processing proteins. DOI: 10.1074/mcp.O113.027870 PMCID: PMC3879628 PMID: 24129315 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/11857738
1. Hum Mutat. 2002 Mar;19(3):225-33. doi: 10.1002/humu.10044. HbVar: A relational database of human hemoglobin variants and thalassemia mutations at the globin gene server. Hardison RC(1), Chui DH, Giardine B, Riemer C, Patrinos GP, Anagnou N, Miller W, Wajcman H. Author information: (1)Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania, USA. We have constructed a relational database of hemoglobin variants and thalassemia mutations, called HbVar, which can be accessed on the web at http://globin.cse.psu.edu. Extensive information is recorded for each variant and mutation, including a description of the variant and associated pathology, hematology, electrophoretic mobility, methods of isolation, stability information, ethnic occurrence, structure studies, functional studies, and references. The initial information was derived from books by Dr. Titus Huisman and colleagues [Huisman et al., 1996, 1997, 1998]. The current database is updated regularly with the addition of new data and corrections to previous data. Queries can be formulated based on fields in the database. Tables of common categories of variants, such as all those involving the alpha1-globin gene (HBA1) or all those that result in high oxygen affinity, are maintained by automated queries on the database. Users can formulate more precise queries, such as identifying "all beta-globin variants associated with instability and found in Scottish populations." This new database should be useful for clinical diagnosis as well as in fundamental studies of hemoglobin biochemistry, globin gene regulation, and human sequence variation at these loci. Copyright 2002 Wiley-Liss, Inc. DOI: 10.1002/humu.10044 PMID: 11857738 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/16798224
1. Ann Thorac Surg. 2006 Jul;82(1):249-53. doi: 10.1016/j.athoracsur.2006.02.033. Inhibition of retinoblastoma tumor suppressor activity by RNA interference in lung cancer lines. Reed MF(1), Zagorski WA, Howington JA, Zilfou JT, Knudsen ES. Author information: (1)Division of Thoracic Surgery, Department of Surgery, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267-0558, USA. michael.reed@uc.edu BACKGROUND: Inactivation of retinoblastoma (RB) tumor suppressor function occurs frequently in lung cancer. Short-hairpin RNA can be constructed to target specific sequences and efficiently knock down protein expression. We developed a short-hairpin RNA approach to specifically target Rb in lung cancer cells to determine the influence of RB knockdown on proliferation. METHODS: NCI-H520 human lung cancer cells (wild-type Rb) were transfected with pMSCVpuro-Rb3C, a plasmid containing a short-hairpin sequence targeted to human Rb. Transfectants harboring the construct were selected with puromycin. Loss of RB expression in selected cell populations was determined by immunoblotting. Proliferating cells were counted to establish growth rates. Retinoblastoma-proficient and RB-deficient tumor growth was monitored in nude mice. RESULTS: Transfection with pMSCVpuro-Rb3C dramatically diminished RB expression and led to aberrant expression of RB-regulated genes. Cells harboring pMSCVpuro-Rb3C grew at an increased rate compared with control cells: 480.6 +/- 37.7 versus 159.4 +/- 36.2 (relative cell count at 12 days). Tumor growth in nude mice also increased with RB knockdown compared with control mice: 135.2 +/- 73.6 mm3 versus 40.0 +/- 17.0 mm3 (tumor volume at 10 days). CONCLUSIONS: Inhibition of RB expression is efficiently achieved in lung cancer cells with short-hairpin RNA. Genetic targets of RB are deregulated with RB knockdown. Retinoblastoma depletion increases growth in vitro and in murine xenografts. These studies indicate that even in the context of an established tumor cell line, RB limits tumorigenic proliferation. Additionally, this model will serve as an ideal system to evaluate the role of RB activity on therapeutic response. DOI: 10.1016/j.athoracsur.2006.02.033 PMID: 16798224 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/22562359
1. Mol Cells. 2012 Jun;33(6):553-62. doi: 10.1007/s10059-012-2269-2. Epub 2012 May 4. Lentiviral vector-mediated shRNA against AIMP2-DX2 suppresses lung cancer cell growth through blocking glucose uptake. Chang SH(1), Chung YS, Hwang SK, Kwon JT, Minai-Tehrani A, Kim S, Park SB, Kim YS, Cho MH. Author information: (1)Laboratory of Toxicology, College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea. Aminoacyl-tRNA synthetases [ARS]-interacting multifunctional protein 2 (AIMP2) has been implicated in the control of cell fate and lung cell differentiation. A variant of AIMP2 lacking exon 2 (AIMP2-DX2) is expressed in different cancer cells. We previously studied the expression level of AIMP2-DX2 in several lung cell lines and reported elevated expression levels of AIMP2-DX2 in NCI-H460 and NCI-H520. Here, we report that the suppression of AIMP2-DX2 by lentivirus mediated short hairpin (sh)RNA (sh-DX2) decreased the rate of glucose uptake and glucose transporters (Gluts) in NCI-H460 cells. Down-regulation of AIMP2-DX2 reduced glycosyltransferase (GnT)-V in the Golgi apparatus, while inducing the GnT-V antagonist GnT-III. Down-regulation of AIMP2-DX2 also suppressed the epidermal growth factor receptor/mitogen activated protein kinase (EGFR/MAPK) signaling pathway, leading to the decrease of the proliferation marker Ki-67 expression in nuclei. Furthermore, dual luciferase activity reduced capdependent protein translation in cells infected with sh-DX2. These results suggest that AIMP2-DX2 may be a relevant therapeutic target for lung cancer, and that the sh-DX2 lentiviral system can be an appropriate method for lung cancer therapy. DOI: 10.1007/s10059-012-2269-2 PMCID: PMC3887752 PMID: 22562359 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/14586397
1. Oncogene. 2003 Oct 30;22(49):7711-5. doi: 10.1038/sj.onc.1207088. Allele-specific patterns of the mouse parathyroid hormone-related protein: influences on cell adhesion and migration. Benelli R(1), Peissel B, Manenti G, Gariboldi M, Vanzetto C, Albini A, Dragani TA. Author information: (1)Molecular Biology Laboratory, Istituto Nazionale per la Ricerca sul Cancro, Genoa, Italy. The mouse parathyroid hormone-like hormone Pthlh(Pro) and Pthlh(Thr) variants are linked with susceptibility and resistance to skin carcinogenesis of Car-S and Car-R mice, respectively, and with in vitro effects (Oncogene, 19: 5324-5328, 2000). We have identified an additional Pthlh variant, consisting of Thr and three amino-acid changes in the C-terminus (Pthlh(SerAspTyr)), carried by an evolutionarily distant Mus spretus (SPRET/Ei) inbred strain. When transfected into NCI-H520 tumor cells, this Pthlh(SerAspTyr) variant did not stimulate tumor growth in nude mice. Analysis of cell adhesion, migration, and invasion patterns of Pthlh(Pro)-, Pthlh(Thr)-, and Pthlh(SerAspTyr)-transfected NCI-H520 cells revealed a 1.5-fold decrease in adhesion efficiency on both collagen type I and Matrigel, and a 5-6-fold increase in migration capability in Pthlh(Pro) transfectants as compared to nontransfected, vector-transfected, Pthlh(Thr)-, or Pthlh(SerAspTyr)-transfected cells. These findings suggest that the cancer modifier effects of the mouse Pthlh gene are mediated by differential cell adhesion and migration effects of PTHrP variants. DOI: 10.1038/sj.onc.1207088 PMID: 14586397 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/22328940
1. PLoS One. 2012;7(2):e31605. doi: 10.1371/journal.pone.0031605. Epub 2012 Feb 6. TET2 mutations are associated with specific 5-methylcytosine and 5-hydroxymethylcytosine profiles in patients with chronic myelomonocytic leukemia. Pérez C(1), Martínez-Calle N, Martín-Subero JI, Segura V, Delabesse E, Fernandez-Mercado M, Garate L, Alvarez S, Rifon J, Varea S, Boultwood J, Wainscoat JS, Cruz Cigudosa J, Calasanz MJ, Cross NC, Prósper F, Agirre X. Author information: (1)Laboratory of Myeloproliferative Syndromes, Oncology Area, University of Navarra, Pamplona, Spain. Chronic myelomonocytic leukemia (CMML) has recently been associated with a high incidence of diverse mutations in genes such as TET2 or EZH2 that are implicated in epigenetic mechanisms. We have performed genome-wide DNA methylation arrays and mutational analysis of TET2, IDH1, IDH2, EZH2 and JAK2 in a group of 24 patients with CMML. 249 genes were differentially methylated between CMML patients and controls. Using Ingenuity pathway analysis, we identified enrichment in a gene network centered around PLC, JNK and ERK suggesting that these pathways, whose deregulation has been recently described in CMML, are affected by epigenetic mechanisms. Mutations of TET2, JAK2 and EZH2 were found in 15 patients (65%), 4 patients (17%) and 1 patient (4%) respectively while no mutations in the IDH1 and IDH2 genes were identified. Interestingly, patients with wild type TET2 clustered separately from patients with TET2 mutations, showed a higher degree of hypermethylation and were associated with higher risk karyotypes. Our results demonstrate the presence of aberrant DNA methylation in CMML and identifies TET2 mutant CMML as a biologically distinct disease subtype with a different epigenetic profile. DOI: 10.1371/journal.pone.0031605 PMCID: PMC3273467 PMID: 22328940 [Indexed for MEDLINE] Conflict of interest statement: Competing Interests: The authors have declared that no competing interests exist.
http://www.ncbi.nlm.nih.gov/pubmed/9099905
1. Mol Cell Endocrinol. 1997 Mar 14;127(1):99-108. doi: 10.1016/s0303-7207(96)03994-9. The noncalcemic vitamin D analogues EB1089 and 22-oxacalcitriol interact with the vitamin D receptor and suppress parathyroid hormone-related peptide gene expression. Falzon M(1). Author information: (1)Department of Pharmacology and Toxicology, and Sealy Center for Molecular Science, The University of Texas Medical Branch, Galveston 77555, USA. Humoral hypercalcemia of malignancy, a frequent complication of squamous cell carcinomas of the lung, is mediated by the parathyroid hormone-related peptide (PTHrP). This study was undertaken to determine whether 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] and two nonhypercalcemic analogues. EB1089 and 22-oxa-1,25(OH)(2)D(3) (OCT), suppress PTHrP gene expression in a human lung squamous cancer cell line, NCI H520. All three compounds (1) decreased steady-state PTHrP mRNA and secreted peptide levels via a transcriptional mechanism; (2) modulated promoter activity of 1,25(OH)(2)D(3)-responsive DNA sequences; and (3) activated the vitamin D receptor (VDR) both in vitro and in vivo. Thus, EB1089 and OCT inhibit PTHrP gene expression in NCI H520 cells and modulate gene expression through the same mechanism as 1,25(OH)(2)D(3), namely, activation of the VDR. 1,25(OH)(2)D(3) is hypercalcemic in vivo. However, the noncalcemic analogues EB1089 and OCT have a therapeutic potential through suppression of PTHrP gene transcription. DOI: 10.1016/s0303-7207(96)03994-9 PMID: 9099905 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/16462760
1. Oncogene. 2006 Jun 29;25(28):3934-8. doi: 10.1038/sj.onc.1209422. Epub 2006 Feb 6. Identification of RASSF8 as a candidate lung tumor suppressor gene. Falvella FS(1), Manenti G, Spinola M, Pignatiello C, Conti B, Pastorino U, Dragani TA. Author information: (1)Department of Experimental Oncology and Laboratories, Istituto Nazionale Tumori, Milan, Italy. The RASSF8 gene, which maps close to the KRAS2 gene, contains a RAS-associated domain and encodes a protein that is evolutionarily conserved from fish to humans. Analysis of the RASSF8 transcript revealed a complex expression pattern of 5'-UTR mRNA isoforms in normal lung and in lung adenocarcinomas (ADCAs), with no apparent differences. However, RASSF8 gene transcript levels were approximately seven-fold-lower in lung ADCAs as compared to normal lung tissue. Expression of RASSF8 protein by transfected lung cancer cells led to inhibition of anchorage-independent growth in soft agar in A549 cells and reduction of clonogenic activity in NCI-H520 cells. These results raise the possibility protein encoded by RASSF8 is a novel tumor suppressor for lung cancer. DOI: 10.1038/sj.onc.1209422 PMID: 16462760 [Indexed for MEDLINE]
http://www.ncbi.nlm.nih.gov/pubmed/12964003
1. Int J Oncol. 2003 Oct;23(4):1187-93. Oridonin induces growth inhibition and apoptosis of a variety of human cancer cells. Ikezoe T(1), Chen SS, Tong XJ, Heber D, Taguchi H, Koeffler HP. Author information: (1)Department of Internal Medicine, Kochi Medical School, Nankoku, Kochi 783-8505, Japan. ikezoet@med.kochi-ms.ac.jp PC-SPES is an eight herbal mixture that was shown to have activity against prostate cancer. Recently, we purified oridonin from Rabdosia rubescens, one component of PC-SPES, by high performance liquid chromatography (HPLC). The ability of oridonin to inhibit the proliferation of cancer cells was examined by MTT assay. Oridonin effectively inhibited the proliferation of a wide variety of cancer cells including those from prostate (LNCaP, DU145, PC3), breast (MCF-7, MDA-MB231), non-small cell lung (NSCL) (NCI-H520, NCI-H460, NCI-H1299) cancers, acute promyelocytic leukemia (NB4), and glioblastoma multiforme (U118, U138) with ED50s ranging from 1.8 to 7.5 micro g/ml. TUNEL assay and cell cycle analysis showed that oridonin induced apoptosis and G0/G1 cell cycle arrest in LNCaP prostate cancer cells. In addition, expression of p21waf1 was induced in LNCaP and NCI-H520 cells in a p53-dependent manner. Interestingly, when p53 was suppressed by over-expression of E6 from human papilloma virus type 16 (HPV-16), these cells lost their sensitivity to oridonin-induced growth inhibition and apoptosis. Taken together, oridonin inhibited the proliferation of cancer cells via apoptosis and cell cycle arrest with p53 playing a central role in several cancer types which express the wild-type p53 gene. Oridonin may be a novel, adjunctive therapy for a large variety of malignancies and probably represents one of the major, active components of PC-SPES. PMID: 12964003 [Indexed for MEDLINE]