The suggested roles of the downstream 3 ' regions acting as a Locus Control Region ( LCR ) , have allowed comparisons to be made between the regulation of the IgH locus with other model systems whose gene expression is governed by LCR activity .

The suggested roles of the <dna>downstream 3 ' regions</dna> acting as a <dna>Locus Control Region</dna> ( <dna>LCR</dna> ) , have allowed comparisons to be made between the regulation of the <dna>IgH locus</dna> with other model systems whose gene expression is governed by <dna>LCR</dna> activity .


Here we summarize the importance of the IgH 3'LCR and its putative functional role in IgH gene expression and compare it with the 5'LCR regulatory region of the human beta-globin locus .

Here we summarize the importance of the <dna>IgH 3'LCR</dna> and its putative functional role in <dna>IgH gene</dna> expression and compare it with the <dna>5'LCR regulatory region</dna> of the <dna>human beta-globin locus</dna> .


HLA binding characteristics and generation of cytotoxic lymphocytes against peptides derived from oncogenic proteins .

HLA binding characteristics and generation of <cell_type>cytotoxic lymphocytes</cell_type> against peptides derived from <protein>oncogenic proteins</protein> .


AIMS AND BACKGROUND : Structurally altered proteins ( derived from chromosomal translocations or gene mutations ) can be considered tumor specific antigens and represent an attractive target for a T-cell mediated response .


T lymphocytes recognize antigens in the form of peptides bound to HLA-molecules .

T lymphocytes recognize antigens in the form of peptides bound to <protein>HLA-molecules</protein> .


MATERIALS AND METHODS : Peptides derived from oncogenic proteins were screened for the presence of HLA binding motifs ; actual binding were evaluated by HLA stabilization experiments using transfectants and specific anti-HLA antibodies .

MATERIALS AND METHODS : Peptides derived from <protein>oncogenic proteins</protein> were screened for the presence of <protein>HLA binding motifs</protein> ; actual binding were evaluated by HLA stabilization experiments using transfectants and specific <protein>anti-HLA antibodies</protein> .


Specific lymphocytes were induced by in vitro peptide sensitization and screened by thymidine uptake or cellular cytotoxic assays .

Specific <cell_type>lymphocytes</cell_type> were induced by in vitro peptide sensitization and screened by thymidine uptake or cellular cytotoxic assays .


RESULTS : We identified peptides derived from EWS/FLI-1 fusion protein and from mutated K-RAS protein ( encompassing respectively the fusion point and the mutation at position 12 ) that showed binding motif for HLA-Cw*0702 and HLA-A3 respectively .

RESULTS : We identified peptides derived from <protein>EWS/FLI-1 fusion protein</protein> and from <protein>mutated K-RAS protein</protein> ( encompassing respectively the <protein>fusion point</protein> and the mutation at <protein>position 12</protein> ) that showed <protein>binding motif</protein> for <protein>HLA-Cw*0702</protein> and <protein>HLA-A3</protein> respectively .


The actual binding of these peptides was analysed in a stabilization assay .


We detected binding for the EWS/FLI-1 peptide and for 5 RAS peptides ( 1 wild type and 4 mutated ) .


The effect of temperature , beta 2-microglobulin ( beta 2-m ) and fetal calf serum ( FCS ) on the binding and the stability of the HLA/peptide complex was studied .

The effect of temperature , <protein>beta 2-microglobulin</protein> ( <protein>beta 2-m</protein> ) and fetal calf serum ( FCS ) on the binding and the stability of the <protein>HLA/peptide complex</protein> was studied .


A low temperature ( 26 degrees C ) increased the binding both in HLA-A3 and HLA-Cw*0702 , while FCS reduced it .

A low temperature ( 26 degrees C ) increased the binding both in <protein>HLA-A3</protein> and <protein>HLA-Cw*0702</protein> , while FCS reduced it .


beta 2-m increased the binding to the HLA-A3 molecule but did not influence the binding to the HLA-Cw*0702 .

<protein>beta 2-m</protein> increased the binding to the <protein>HLA-A3 molecule</protein> but did not influence the binding to the <protein>HLA-Cw*0702</protein> .


The stability of already formed complexed was somewhat different in the HLA-A3 and HLA-Cw*0702 system : both were more stable at 26 degrees C than at 37 degrees C but while the beta 2-m and FCS did not influence the stability of the HLA-A3/peptide complex , they seemed to cause opposite effects in the HLA-Cw*0702 system ( beta 2-m stabilized and FCS destabilized the complex ) .

The stability of already formed complexed was somewhat different in the <protein>HLA-A3</protein> and <protein>HLA-Cw*0702</protein> system : both were more stable at 26 degrees C than at 37 degrees C but while the <protein>beta 2-m</protein> and FCS did not influence the stability of the <protein>HLA-A3/peptide complex</protein> , they seemed to cause opposite effects in the <protein>HLA-Cw*0702</protein> system ( <protein>beta 2-m</protein> stabilized and FCS destabilized the complex ) .


Finally , we were able to generate a specific CD8+ CTL line against a K-RAS mutated peptide .

Finally , we were able to generate a specific <cell_line>CD8+ CTL line</cell_line> against a K-RAS mutated peptide .


CONCLUSIONS : Although binding motifs and actual HLA binding can be detected in several cases , the generation of a cellular response is infrequent , confirming that HLA binding is necessary but not sufficient to obtain an in vitro response .

CONCLUSIONS : Although <protein>binding motifs</protein> and actual HLA binding can be detected in several cases , the generation of a cellular response is infrequent , confirming that HLA binding is necessary but not sufficient to obtain an in vitro response .


Further optimization of culture conditions , type of Antigen Presenting Cells ( APC ) , peptides , use of stabilizers like beta 2-m are still needed .

Further optimization of culture conditions , type of <cell_type>Antigen Presenting Cells</cell_type> ( <cell_type>APC</cell_type> ) , peptides , use of stabilizers like <protein>beta 2-m</protein> are still needed .


OCA-B is a functional analog of VP16 but targets a separate surface of the Oct-1 POU domain [ published erratum appears in Mol Cell Biol 1998 Apr ; 18 ( 4 ) : 2430 ]

<protein>OCA-B</protein> is a functional analog of <protein>VP16</protein> but targets a separate surface of the <protein>Oct-1 POU domain</protein> [ published erratum appears in Mol Cell Biol 1998 Apr ; 18 ( 4 ) : 2430 ]


OCA-B is a B-cell-specific coregulator of the broadly expressed POU domain transcription factor Oct-1 .

<protein>OCA-B</protein> is a <protein>B-cell-specific coregulator</protein> of the broadly expressed <protein>POU domain transcription factor</protein> <protein>Oct-1</protein> .


OCA-B associates with the Oct-1 POU domain , a bipartite DNA-binding structure containing a POU-specific ( POU [ S ] ) domain joined by a flexible linker to a POU homeodomain ( POU [ H ] ) .

<protein>OCA-B</protein> associates with the <protein>Oct-1 POU domain</protein> , a bipartite DNA-binding structure containing a <protein>POU-specific ( POU [ S ] ) domain</protein> joined by a flexible linker to a <protein>POU homeodomain</protein> ( <protein>POU [ H ]</protein> ) .


Here , we show that OCA-B alters the activity of Oct-1 in two ways .

Here , we show that <protein>OCA-B</protein> alters the activity of <protein>Oct-1</protein> in two ways .


It provides a transcriptional activation domain which , unlike Oct-1 , activates an mRNA-type promoter effectively , and it stabilizes Oct-1 on the Oct-1-responsive octamer sequence ATGCAAAT .

It provides a transcriptional activation domain which , unlike <protein>Oct-1</protein> , activates an <dna>mRNA-type promoter</dna> effectively , and it stabilizes <protein>Oct-1</protein> on the <dna>Oct-1-responsive octamer sequence</dna> ATGCAAAT .


These properties of OCA-B parallel those displayed by the herpes simplex virus Oct-1 coregulator VP16 .

These properties of <protein>OCA-B</protein> parallel those displayed by the <protein>herpes simplex virus Oct-1 coregulator</protein> <protein>VP16</protein> .


OCA-B , however , interacts with a different surface of the DNA-bound Oct-1 POU domain , interacting with both the POU ( S ) and POU ( H ) domains and the center of the ATGCAAAT octamer sequence .

<protein>OCA-B</protein> , however , interacts with a different surface of the DNA-bound <protein>Oct-1 POU domain</protein> , interacting with both the <protein>POU ( S ) and POU ( H ) domains</protein> and the center of the <dna>ATGCAAAT octamer sequence</dna> .


The OCA-B and VP16 interactions with the Oct-1 POU domain are sufficiently different to permit OCA-B and VP16 to bind the Oct-1 POU domain simultaneously .

The OCA-B and <protein>VP16</protein> interactions with the <protein>Oct-1 POU domain</protein> are sufficiently different to permit <protein>OCA-B</protein> and <protein>VP16</protein> to bind the <protein>Oct-1 POU domain</protein> simultaneously .


These results emphasize the structural versatility of the Oct-1 POU domain in its interaction with coregulators .

These results emphasize the structural versatility of the <protein>Oct-1 POU domain</protein> in its interaction with coregulators .


Phosphatidylinositol 3-kinase couples the interleukin-2 receptor to the cell cycle regulator E2F .

<protein>Phosphatidylinositol 3-kinase</protein> couples the <protein>interleukin-2 receptor</protein> to the <protein>cell cycle regulator</protein> <protein>E2F</protein> .


Cell cycle progression initiated by interleukin-2 ( IL-2 ) in T cells is critical for lymphoproliferation and an immune response .

Cell cycle progression initiated by <protein>interleukin-2</protein> ( <protein>IL-2</protein> ) in <cell_type>T cells</cell_type> is critical for lymphoproliferation and an immune response .


Phosphatidyl inositol 3-kinase ( PI3K ) is activated by IL-2 .

<protein>Phosphatidyl inositol 3-kinase</protein> ( <protein>PI3K</protein> ) is activated by <protein>IL-2</protein> .


However , nuclear targets for PI3K are not known .

However , nuclear targets for <protein>PI3K</protein> are not known .


Here we identify the cell cycle regulator E2F as an IL-2 target in T lymphocytes and PI3K as the critical signaling pathway .

Here we identify the <protein>cell cycle regulator</protein> <protein>E2F</protein> as an <protein>IL-2</protein> target in T <cell_type>lymphocytes</cell_type> and <protein>PI3K</protein> as the critical signaling pathway .


We eliminate both Stat5 and Raf /MEK pathways from E2F regulation .

We eliminate both <protein>Stat5</protein> and <protein>Raf</protein> <protein>/MEK</protein> pathways from <protein>E2F</protein> regulation .


Protein kinase B ( PKB ) is activated by IL-2 via PI3K .

<protein>Protein kinase B</protein> ( <protein>PKB</protein> ) is activated by <protein>IL-2</protein> via <protein>PI3K</protein> .


The expression of an active PKB is sufficient to induce E2F activity .

The expression of an active <protein>PKB</protein> is sufficient to induce <protein>E2F</protein> activity .


Inhibition of PI3K inhibits phosphorylation of Rb , induction of cyclin D3 , and degradation of p27kip1 .

Inhibition of <protein>PI3K</protein> inhibits phosphorylation of <protein>Rb</protein> , induction of <protein>cyclin D3</protein> , and degradation of <protein>p27kip1</protein> .


These results establish a crucial PI3K / PKB -mediated link between the IL-2 teceptor and the cell cycle machinery .

These results establish a crucial <protein>PI3K</protein> / <protein>PKB</protein> -mediated link between the <protein>IL-2 teceptor</protein> and the cell cycle machinery .


The Epstein-Barr virus oncogene product latent membrane protein 1 engages the tumor necrosis factor receptor-associated death domain protein to mediate B lymphocyte growth transformation and activate NF-kappaB .

The <protein>Epstein-Barr virus oncogene product</protein> <protein>latent membrane protein 1</protein> engages the tumor necrosis factor receptor-associated death domain protein to mediate B lymphocyte growth transformation and activate NF-kappaB .


The Epstein-Barr virus latent membrane protein 1 ( LMP1 ) is essential for the transformation of B lymphocytes into lymphoblastoid cell lines .

The Epstein-Barr virus <protein>latent membrane protein 1</protein> ( <protein>LMP1</protein> ) is essential for the transformation of <cell_type>B lymphocytes</cell_type> into <cell_line>lymphoblastoid cell lines</cell_line> .


Previous data are consistent with a model that LMP1 is a constitutively activated receptor that transduces signals for transformation through its carboxyl-terminal cytoplasmic tail .

Previous data are consistent with a model that <protein>LMP1</protein> is a <protein>constitutively activated receptor</protein> that transduces signals for transformation through its <protein>carboxyl-terminal cytoplasmic tail</protein> .


One transformation effector site ( TES1 ) , located within the membrane proximal 45 residues of the cytoplasmic tail , constitutively engages tumor necrosis factor receptor-associated factors .

One <protein>transformation effector site</protein> ( <protein>TES1</protein> ) , located within the <protein>membrane proximal 45 residues</protein> of the cytoplasmic tail , constitutively engages <protein>tumor necrosis factor receptor-associated factors</protein> .


Signals from TES1 are sufficient to drive initial proliferation of infected resting B lymphocytes , but most lymphoblastoid cells infected with a virus that does not express the 155 residues beyond TES1 fail to grow as long-term cell lines .

Signals from <protein>TES1</protein> are sufficient to drive initial proliferation of infected resting B <cell_type>lymphocytes</cell_type> , but most <cell_type>lymphoblastoid cells</cell_type> infected with a virus that does not express the <protein>155 residues</protein> beyond <protein>TES1</protein> fail to grow as <cell_line>long-term cell lines</cell_line> .


We now find that mutating two tyrosines to an isoleucine at the carboxyl end of the cytoplasmic tail cripples the ability of EBV to cause lymphoblastoid cell outgrowth , thereby marking a second transformation effector site , TES2 .

We now find that mutating two tyrosines to an isoleucine at the <protein>carboxyl end</protein> of the <protein>cytoplasmic tail</protein> cripples the ability of EBV to cause lymphoblastoid cell outgrowth , thereby marking a second <protein>transformation effector site</protein> , <protein>TES2</protein> .


A yeast two-hybrid screen identified TES2 interacting proteins , including the tumor necrosis factor receptor-associated death domain protein ( TRADD ) .

A yeast two-hybrid screen identified <protein>TES2 interacting proteins</protein> , including the <protein>tumor necrosis factor receptor-associated death domain protein</protein> ( <protein>TRADD</protein> ) .


TRADD was the only protein that interacted with wild-type TES2 and not with isoleucine-mutated TES2 .

<protein>TRADD</protein> was the only protein that interacted with <protein>wild-type TES2</protein> and not with <protein>isoleucine-mutated TES2</protein> .


TRADD associated with wild-type LMP1 but not with isoleucine-mutated LMP1 in mammalian cells , and TRADD constitutively associated with LMP1 in EBV-transformed cells .

<protein>TRADD</protein> associated with <protein>wild-type LMP1</protein> but not with isoleucine-mutated <protein>LMP1</protein> in mammalian cells , and <protein>TRADD</protein> constitutively associated with <protein>LMP1</protein> in <cell_line>EBV-transformed cells</cell_line> .


In transfection assays , TRADD and TES2 synergistically mediated high-level NF-kappaB activation .

In transfection assays , <protein>TRADD</protein> and <protein>TES2</protein> synergistically mediated high-level <protein>NF-kappaB</protein> activation .


These results indicate that LMP1 appropriates TRADD to enable efficient long-term lymphoblastoid cell outgrowth .

These results indicate that <protein>LMP1</protein> appropriates <protein>TRADD</protein> to enable efficient long-term lymphoblastoid cell outgrowth .


High-level NF-kappaB activation also appears to be a critical component of long-term outgrowth .

High-level <protein>NF-kappaB</protein> activation also appears to be a critical component of long-term outgrowth .


Inhibition of T cell and promotion of natural killer cell development by the dominant negative helix loop helix factor Id3 .

Inhibition of <cell_type>T cell</cell_type> and promotion of natural killer cell development by the <protein>dominant negative helix loop helix factor</protein> <protein>Id3</protein> .


Bipotential T/natural killer ( NK ) progenitor cells are present in the human thymus .

<cell_type>Bipotential T/natural killer ( NK ) progenitor cells</cell_type> are present in the human thymus .


Despite their bipotential capacity , these progenitors develop predominantly to T cells in the thymus .

Despite their bipotential capacity , these progenitors develop predominantly to <cell_type>T cells</cell_type> in the thymus .


The mechanisms controlling this developmental choice are unknown .


Here we present evidence that a member ( s ) of the family of basic helix loop helix ( bHLH ) transcription factors determines lineage specification of NK/T cell progenitors .

Here we present evidence that a member ( s ) of the family of <protein>basic helix loop helix ( bHLH ) transcription factors</protein> determines lineage specification of <cell_type>NK/T cell progenitors</cell_type> .


The natural dominant negative HLH factor Id3 , which blocks transcriptional activity of a number of known bHLH factors , was expressed in CD34+ progenitor cells by retrovirus-mediated gene transfer .

The natural <protein>dominant negative HLH factor</protein> <protein>Id3</protein> , which blocks transcriptional activity of a number of known <protein>bHLH factors</protein> , was expressed in <cell_type>CD34+ progenitor cells</cell_type> by retrovirus-mediated gene transfer .


Constitutive expression of Id3 completely blocks development of CD34+ cells into T cells in a fetal thymic organ culture ( FTOC ) .

Constitutive expression of <protein>Id3</protein> completely blocks development of <cell_type>CD34+ cells</cell_type> into <cell_type>T cells</cell_type> in a fetal thymic organ culture ( FTOC ) .


In contrast , development into NK cells in an FTOC is enhanced .

In contrast , development into <cell_type>NK cells</cell_type> in an FTOC is enhanced .


Thus , the activity of a bHLH transcription factor is necessary for T lineage differentiation of bipotential precursors , in the absence of which a default pathway leading to NK cell development is chosen .

Thus , the activity of a <protein>bHLH transcription factor</protein> is necessary for T lineage differentiation of <cell_type>bipotential precursors</cell_type> , in the absence of which a default pathway leading to NK cell development is chosen .


Our results identify a molecular switch for lineage specification in early lymphoid precursors of humans .

Our results identify a molecular switch for lineage specification in <cell_type>early lymphoid precursors</cell_type> of humans .


Late gene expression from the Epstein-Barr virus BcLF1 and BFRF3 promoters does not require DNA replication in cis .

Late gene expression from the Epstein-Barr virus <dna>BcLF1 and BFRF3 promoters</dna> does not require DNA replication in cis .


Late gene expression follows and is dependent upon lytic replication of the viral genome .

Late gene expression follows and is dependent upon lytic replication of the <dna>viral genome</dna> .


Although experimental evidence is lacking , lytic viral DNA replication is believed to remove modifications or binding factors from the genome which serve to repress late gene expression during latency or the early lytic cycle .

Although experimental evidence is lacking , lytic viral DNA replication is believed to remove modifications or <protein>binding factors</protein> from the genome which serve to repress late gene expression during latency or the early lytic cycle .


We have developed a reporter assay to begin characterizing the mechanisms that regulate late gene expression in Epstein-Barr virus ( EBV ) .


In this model system , the activities of late promoter-reporter fusions are measured following transient transfection into tissue culture cells expressing EBV during different stages of the lytic cycle .

In this model system , the activities of <dna>late promoter-reporter fusions</dna> are measured following transient transfection into <cell_line>tissue culture cells</cell_line> expressing EBV during different stages of the lytic cycle .


This system faithfully recapitulates late expression patterns from the endogenous virus , implicating specific cis-active sequences in the control of late gene expression .

This system faithfully recapitulates late expression patterns from the endogenous virus , implicating specific <dna>cis-active sequences</dna> in the control of late gene expression .


In addition , these promoters respond only indirectly to the viral immediate-early transactivator , ZEBRA .

In addition , these promoters respond only indirectly to the <dna>viral immediate-early transactivator</dna> , <dna>ZEBRA</dna> .


This indirect response is mediated by other viral or virally induced activities downstream of ZEBRA in the lytic cascade .

This indirect response is mediated by other viral or virally induced activities downstream of <dna>ZEBRA</dna> in the lytic cascade .


In this system , late gene expression is sensitive to inhibitors of the viral DNA polymerase such as phosphonoacetic acid , although the reporters lack a eukaryotic origin of replication and are not replicated under the assay conditions .

In this system , late gene expression is sensitive to inhibitors of the <protein>viral DNA polymerase</protein> such as phosphonoacetic acid , although the reporters lack a eukaryotic origin of replication and are not replicated under the assay conditions .


Thus , replication of the transcriptional template is not a prerequisite for expression with late kinetics , a finding inconsistent with the current models which posit a cis-active relationship between lytic EBV DNA replication and late gene expression .

Thus , replication of the <dna>transcriptional template</dna> is not a prerequisite for expression with late kinetics , a finding inconsistent with the current models which posit a cis-active relationship between lytic EBV DNA replication and late gene expression .


Rather , analysis of this system has revealed a trans relationship between late gene expression and viral DNA replication and highlights the indirect and complex link between these two events .


Modulation of mRNA expression of a novel human myeloid-selective CCAAT/enhancer binding protein gene ( C/EBP epsilon ) .

Modulation of mRNA expression of a <dna>novel human myeloid-selective CCAAT/enhancer binding protein gene</dna> ( <dna>C/EBP epsilon</dna> ) .


Human C/EBP epsilon is a newly cloned gene coding for a CCAAT/enhancer binding protein that may be involved in the regulation of myeloid differentiation .

Human <dna>C/EBP epsilon</dna> is a newly cloned gene coding for a <protein>CCAAT/enhancer binding protein</protein> that may be involved in the regulation of myeloid differentiation .


Our studies showed that levels of C/EBP epsilon mRNA were markedly increased in NB4 cells ( promyelocytic leukemia line ) , because they were induced by 9-cis retinoic acid ( 9-cis RA ) to differentiate towards granulocytes .

Our studies showed that levels of <rna>C/EBP epsilon mRNA</rna> were markedly increased in <cell_line>NB4 cells</cell_line> ( <cell_line>promyelocytic leukemia line</cell_line> ) , because they were induced by 9-cis retinoic acid ( 9-cis RA ) to differentiate towards <cell_type>granulocytes</cell_type> .


Accumulation of C/EBP epsilon mRNA occurred as early as 1 hour after exposure of NB4 cells to 9-cis RA ( 5 x 10 ( -7 ) mol/L ) ; and at 48 hours , levels were increased by 5.1-fold .

Accumulation of <rna>C/EBP epsilon mRNA</rna> occurred as early as 1 hour after exposure of <cell_line>NB4 cells</cell_line> to 9-cis RA ( 5 x 10 ( -7 ) mol/L ) ; and at 48 hours , levels were increased by 5.1-fold .


Dose-response studies showed that 10 ( -7 ) to 10 ( -6 ) mol/L 9-cis RA ( 12 hours ) resulted in peak levels of C/EBP epsilon mRNA ; but even 10 ( -10 ) mol/L 9-cis RA increased levels of these transcripts .

Dose-response studies showed that 10 ( -7 ) to 10 ( -6 ) mol/L 9-cis RA ( 12 hours ) resulted in peak levels of <rna>C/EBP epsilon mRNA</rna> ; but even 10 ( -10 ) mol/L 9-cis RA increased levels of these transcripts .


NB4 cells pulse-exposed ( 30 minutes ) to all-trans retinoic acid ( ATRA ) , washed , and cultured ( 3 days ) with either dimethylsulfoxide ( DMSO ) or hexamethylene bisacetamide ( HMBA ) had a prominent increase in levels of C/EBP epsilon mRNA and an increase in granulocytic differentiation , but exposure to either DMSO or HMBA alone had no effect on base levels of C/EBP epsilon and did not induce differentiation .

<cell_line>NB4 cells</cell_line> pulse-exposed ( 30 minutes ) to all-trans retinoic acid ( ATRA ) , washed , and cultured ( 3 days ) with either dimethylsulfoxide ( DMSO ) or hexamethylene bisacetamide ( HMBA ) had a prominent increase in levels of <rna>C/EBP epsilon mRNA</rna> and an increase in granulocytic differentiation , but exposure to either DMSO or HMBA alone had no effect on base levels of C/EBP epsilon and did not induce differentiation .


Macrophage-differentiation of NB4 reduced levels of C/EBP epsilon mRNA .

Macrophage-differentiation of <cell_line>NB4</cell_line> reduced levels of <rna>C/EBP epsilon mRNA</rna> .


Nuclear run-off assays and half-life studies showed that accumulation of C/EBP epsilon mRNA by 9-cis RA was due to enhanced transcription .

Nuclear run-off assays and half-life studies showed that accumulation of <rna>C/EBP epsilon mRNA</rna> by 9-cis RA was due to enhanced transcription .


Furthermore , this C/EBP epsilon mRNA accumulation did not require synthesis of new protein factors because 9-cis RA induced C/EBP epsilon mRNA accumulation in the absence of new protein synthesis .

Furthermore , this <rna>C/EBP epsilon mRNA</rna> accumulation did not require synthesis of new <protein>protein factors</protein> because 9-cis RA induced <rna>C/EBP epsilon mRNA</rna> accumulation in the absence of new protein synthesis .


ATRA also induced expression of C/EBP epsilon protein in NB4 cells , as shown by Western blotting .

ATRA also induced expression of <protein>C/EBP epsilon protein</protein> in <cell_line>NB4 cells</cell_line> , as shown by Western blotting .


In contrast to the increase of C/EBP epsilon in 9-cis RA-mediated granulocytic differentiation , the DMSO-induced differentiation of HL-60 cells down the granulocytic pathway was associated with an initial reduction of C/EBP epsilon mRNA levels .

In contrast to the increase of C/EBP epsilon in 9-cis RA-mediated granulocytic differentiation , the DMSO-induced differentiation of <cell_line>HL-60 cells</cell_line> down the granulocytic pathway was associated with an initial reduction of <rna>C/EBP epsilon mRNA</rna> levels .


In summary , we have discovered that expression of C/EBP epsilon mRNA is markedly enhanced as the NB4 promyelocytes are induced by retinoids to differentiate towards granulocytes .

In summary , we have discovered that expression of <rna>C/EBP epsilon mRNA</rna> is markedly enhanced as the <cell_line>NB4 promyelocytes</cell_line> are induced by retinoids to differentiate towards <cell_type>granulocytes</cell_type> .


This induction of C/EBP epsilon mRNA expression is transcriptionally mediated and occurs in the absence of synthesis of additional protein factors .

This induction of <rna>C/EBP epsilon mRNA</rna> expression is transcriptionally mediated and occurs in the absence of synthesis of additional <protein>protein factors</protein> .


We suspect that the C/EBP epsilon promoter/enhancer contains a retinoic acid-response element that is directly stimulated by retinoids .

We suspect that the <dna>C/EBP epsilon promoter/enhancer</dna> contains a <dna>retinoic acid-response element</dna> that is directly stimulated by retinoids .


Dual effects of LPS antibodies on cellular uptake of LPS and LPS-induced proinflammatory functions .

Dual effects of <protein>LPS antibodies</protein> on cellular uptake of LPS and LPS-induced proinflammatory functions .


Human phagocytes recognize bacterial LPS ( endotoxin ) through membrane CD14 ( mCD14 ) , a proinflammatory LPS receptor .

<cell_type>Human phagocytes</cell_type> recognize bacterial LPS ( endotoxin ) through <protein>membrane CD14</protein> ( <protein>mCD14</protein> ) , a <protein>proinflammatory LPS receptor</protein> .


This study tested the hypothesis that anti-LPS Abs neutralize endotoxin by blocking cellular uptake through mCD14 .

This study tested the hypothesis that <protein>anti-LPS Abs</protein> neutralize endotoxin by blocking cellular uptake through <protein>mCD14</protein> .


Ab-associated changes in the uptake and cellular distribution of FITC-LPS were assessed by flow cytometry and laser scanning confocal microscopy in human CD14-transfected Chinese hamster ovary fibroblasts ( CHO-CD14 cells ) and human peripheral blood monocytes .

Ab-associated changes in the uptake and cellular distribution of FITC-LPS were assessed by flow cytometry and laser scanning confocal microscopy in <cell_line>human CD14-transfected Chinese hamster ovary fibroblasts</cell_line> ( <cell_line>CHO-CD14 cells</cell_line> ) and <cell_type>human peripheral blood monocytes</cell_type> .


LPS core- and O-side chain-specific mAbs inhibited mCD14 -mediated LPS uptake by both cell types in the presence of serum .

LPS <protein>core- and O-side chain-specific mAbs</protein> inhibited <protein>mCD14</protein> -mediated LPS uptake by both cell types in the presence of serum .


O-side chain-specific mAb concurrently enhanced complement-dependent LPS uptake by monocytes through complement receptor-1 ( CR1 ) and uptake by CHO-CD14 cells involving another heat-labile serum factor ( s ) and cell-associated recognition molecule ( s ) .

<protein>O-side chain-specific mAb</protein> concurrently enhanced complement-dependent LPS uptake by monocytes through <protein>complement receptor-1</protein> ( <protein>CR1</protein> ) and uptake by <cell_line>CHO-CD14 cells</cell_line> involving another <protein>heat-labile serum factor</protein> ( s ) and <protein>cell-associated recognition molecule</protein> ( s ) .


Core-specific mAb inhibited mCD14 -mediated uptake of homologous and heterologous LPS , while producing less concurrent enhancement of non- mCD14 -mediated LPS uptake .

<protein>Core-specific mAb</protein> inhibited <protein>mCD14</protein> -mediated uptake of homologous and heterologous LPS , while producing less concurrent enhancement of non- <protein>mCD14</protein> -mediated LPS uptake .


The modulation by anti-LPS mAbs of mCD14 -mediated LPS uptake was associated with inhibition of LPS-induced nuclear factor-kappaB ( NF-kappaB ) translocation and TNF-alpha secretion in CHO-CD14 cells and monocytes , respectively , while mAb enhancement of non- mCD14 -mediated LPS uptake stimulated these activities .

The modulation by <protein>anti-LPS mAbs</protein> of <protein>mCD14</protein> -mediated LPS uptake was associated with inhibition of <protein>LPS-induced nuclear factor-kappaB</protein> ( <protein>NF-kappaB</protein> ) translocation and <protein>TNF-alpha</protein> secretion in <cell_line>CHO-CD14 cells</cell_line> and monocytes , respectively , while mAb enhancement of non- <protein>mCD14</protein> -mediated LPS uptake stimulated these activities .


LPS-specific Abs thus mediate anti-inflammatory and proinflammatory functions , respectively , by preventing target cell uptake of LPS through mCD14 and augmenting uptake through CR1 or other cell receptors .

<protein>LPS-specific Abs</protein> thus mediate anti-inflammatory and proinflammatory functions , respectively , by preventing target cell uptake of LPS through <protein>mCD14</protein> and augmenting uptake through <protein>CR1</protein> or other <protein>cell receptors</protein> .


Four P-like elements are required for optimal transcription of the mouse IL-4 gene : involvement of a distinct set of nuclear factor of activated T cells and activator protein-1 family proteins .

Four <dna>P-like elements</dna> are required for optimal transcription of the <dna>mouse IL-4 gene</dna> : involvement of a distinct set of <protein>nuclear factor of activated T cells</protein> and <protein>activator protein-1 family proteins</protein> .


We previously identified the P sequence as a critical regulatory element of the human IL-4 promoter .

We previously identified the <dna>P sequence</dna> as a critical <dna>regulatory element</dna> of the <dna>human IL-4 promoter</dna> .


In the mouse IL-4 promoter , there are five elements homologous to the human P sequence designated conserved lymphokine element 0 ( CLE0 ) , P , P2 , P3 and P4 .

In the <dna>mouse IL-4 promoter</dna> , there are five elements homologous to the human <dna>P sequence</dna> designated <dna>conserved lymphokine element 0</dna> ( <dna>CLE0</dna> ) , <dna>P</dna> , <dna>P2</dna> , <dna>P3</dna> and <dna>P4</dna> .


To characterize the role of these P-like elements and their binding factors in the native promoter , we did transient transfection and electrophoretic mobility shift assays ( EMSA ) .

To characterize the role of these <dna>P-like elements</dna> and their <protein>binding factors</protein> in the <dna>native promoter</dna> , we did transient transfection and electrophoretic mobility shift assays ( EMSA ) .


Transfection of EL-4 cells with the IL-4 promoter-reporter constructs carrying mutated P-like elements showed that four P-like elements , CLE0 , P , P2 and P4 , but not P3 , were required for optimal activation of the IL-4 promoter .

Transfection of <cell_line>EL-4 cells</cell_line> with the <dna>IL-4 promoter-reporter constructs</dna> carrying <dna>mutated P-like elements</dna> showed that four <dna>P-like elements</dna> , <dna>CLE0</dna> , <dna>P</dna> , <dna>P2</dna> and <dna>P4</dna> , but not <dna>P3</dna> , were required for optimal activation of the <dna>IL-4 promoter</dna> .


EMSA showed that both constitutive and inducible complexes bound to CLE0 , P , P2 and P4 , whereas only a constitutive complex bound to P3 .

EMSA showed that both constitutive and inducible complexes bound to <dna>CLE0</dna> , <dna>P</dna> , <dna>P2</dna> and <dna>P4</dna> , whereas only a constitutive complex bound to <dna>P3</dna> .


In competition and antibody supershift assays in EMSA , complexes formed with P or P2 proved to contain nuclear factor of activated T cells ( NFAT ) family proteins as major components .

In competition and antibody supershift assays in EMSA , complexes formed with <dna>P</dna> or <dna>P2</dna> proved to contain <protein>nuclear factor of activated T cells ( NFAT ) family proteins</protein> as major components .


Activator protein ( AP ) -1 family proteins interacted with CLE0 , P , P2 and P4 .

<protein>Activator protein ( AP ) -1 family proteins</protein> interacted with <dna>CLE0</dna> , P , <dna>P2</dna> and <dna>P4</dna> .


NFAT/AP-1 complex formed only with P and P2 .

<protein>NFAT/AP-1 complex</protein> formed only with <dna>P</dna> and <dna>P2</dna> .


Cross-competition assays among the P-like elements revealed element-specific and common complexes .

Cross-competition assays among the <dna>P-like elements</dna> revealed <protein>element-specific and common complexes</protein> .


Six tandem repeats of the P element linked to the SV40 promoter responded to phorbol 12-myristate 13-acetate , while that of other elements did not. It would thus appear that components of each P-like element-binding complexes are not identical and may coordinately contribute to transcriptional activity .

Six tandem repeats of the <dna>P element</dna> linked to the <dna>SV40 promoter</dna> responded to phorbol 12-myristate 13-acetate , while that of other elements did not. It would thus appear that components of each <protein>P-like element-binding complexes</protein> are not identical and may coordinately contribute to transcriptional activity .


Constitutive expression c-fos , c-jun , and NF kappa B mRNA is in nucleated fetal blood cells and up-regulation of c-fos and c-jun with anti-CD3 stimulation .

Constitutive expression <rna>c-fos , c-jun , and NF kappa B mRNA</rna> is in <cell_type>nucleated fetal blood cells</cell_type> and up-regulation of <protein>c-fos</protein> and <protein>c-jun</protein> with <protein>anti-CD3</protein> stimulation .


Fetal and neonatal lymphocytes are relatively resistant to activation and cytokine production when stimulated either via their T-cell antigen receptors or lectins .

<cell_type>Fetal and neonatal lymphocytes</cell_type> are relatively resistant to activation and <protein>cytokine</protein> production when stimulated either via their <protein>T-cell antigen receptors</protein> or <protein>lectins</protein> .


The molecular mechanism ( s ) responsible for this phenomenon have not been clearly elucidated .


We have hypothesized that such defects in fetal/neonatal T-cell activation may be due to lack of expression of the transcriptional regulatory elements required for T-cell activation .

We have hypothesized that such defects in fetal/neonatal T-cell activation may be due to lack of expression of the <protein>transcriptional regulatory elements</protein> required for T-cell activation .


We used reverse transcriptase -polymerase chain reaction to examine both fetal and term neonatal cord bloods for mRNA expression of three transcription factors implicated in T-cell activation : c-jun , c-fos , and NF kappa B ( p50 subunit ) .

We used <protein>reverse transcriptase</protein> -polymerase chain reaction to examine both fetal and term neonatal cord bloods for mRNA expression of three <protein>transcription factors</protein> implicated in T-cell activation : <protein>c-jun</protein> , <protein>c-fos</protein> , and <protein>NF kappa B</protein> ( <protein>p50 subunit</protein> ) .


We demonstrate that mRNAs for all three of these regulatory factors are expressed in fetal blood cells by the 27th week of gestation and in term cord bloods .

We demonstrate that <rna>mRNAs</rna> for all three of these regulatory factors are expressed in <cell_type>fetal blood cells</cell_type> by the 27th week of gestation and in term cord bloods .


Activation of term infant cord blood mononuclear cells with anti-CD3 monoclonal antibodies resulted in up-regulation of both c-jun and c-fos mRNAs within 15 min of stimulation .

Activation of <cell_type>term infant cord blood mononuclear cells</cell_type> with <protein>anti-CD3 monoclonal antibodies</protein> resulted in up-regulation of both <protein>c-jun</protein> and c-fos <rna>mRNAs</rna> within 15 min of stimulation .


However , secretion of IL-2 by anti-CD3-stimulated cord blood mononuclear cells was still blunted compared with control cells from adults .

However , secretion of <protein>IL-2</protein> by <cell_line>anti-CD3-stimulated cord blood mononuclear cells</cell_line> was still blunted compared with <cell_type>control cells</cell_type> from adults .


We conclude that fetal nucleated blood cells constitutively express important genes for cytokine regulation and are able to increase intracellular accumulation of the mRNAs for these factors in response to anti-CD3 stimulation .

We conclude that <cell_type>fetal nucleated blood cells</cell_type> constitutively express important genes for <protein>cytokine</protein> regulation and are able to increase intracellular accumulation of the <rna>mRNAs</rna> for these factors in response to <protein>anti-CD3</protein> stimulation .


Thus , qualitative differences in the capacity to regulate these factors could not be shown in fetal blood cells .

Thus , qualitative differences in the capacity to regulate these factors could not be shown in <cell_type>fetal blood cells</cell_type> .


Quantitative experiments comparing binding of these transcription factors to the IL-2 promoter are currently under investigation .

Quantitative experiments comparing binding of these <protein>transcription factors</protein> to the <dna>IL-2 promoter</dna> are currently under investigation .


Nuclear accumulation of NFAT4 opposed by the JNK signal transduction pathway .

Nuclear accumulation of <protein>NFAT4</protein> opposed by the <protein>JNK</protein> signal transduction pathway .


The nuclear factor of activated T cells ( NFAT ) group of transcription factors is retained in the cytoplasm of quiescent cells .

The <protein>nuclear factor of activated T cells ( NFAT ) group</protein> of <protein>transcription factors</protein> is retained in the cytoplasm of <cell_type>quiescent cells</cell_type> .


NFAT activation is mediated in part by induced nuclear import .

<protein>NFAT</protein> activation is mediated in part by induced nuclear import .


This process requires calcium-dependent dephosphorylation of NFAT caused by the phosphatase calcineurin .

This process requires calcium-dependent dephosphorylation of <protein>NFAT</protein> caused by the <protein>phosphatase</protein> <protein>calcineurin</protein> .


The c-Jun amino-terminal kinase ( JNK ) phosphorylates NFAT4 on two sites .

The <protein>c-Jun amino-terminal kinase</protein> ( <protein>JNK</protein> ) phosphorylates <protein>NFAT4</protein> on two sites .


Mutational removal of the JNK phosphorylation sites caused constitutive nuclear localization of NFAT4 .

Mutational removal of the <protein>JNK phosphorylation sites</protein> caused constitutive nuclear localization of <protein>NFAT4</protein> .


In contrast , JNK activation in calcineurin-stimulated cells caused nuclear exclusion of NFAT4 .

In contrast , <protein>JNK</protein> activation in <cell_line>calcineurin-stimulated cells</cell_line> caused nuclear exclusion of <protein>NFAT4</protein> .


These findings show that the nuclear accumulation of NFAT4 promoted by calcineurin is opposed by the JNK signal transduction pathway .

These findings show that the nuclear accumulation of <protein>NFAT4</protein> promoted by <protein>calcineurin</protein> is opposed by the <protein>JNK</protein> signal transduction pathway .


Thiol modulation inhibits the interleukin ( IL ) -1 -mediated activation of an IL-1 receptor-associated protein kinase and NF-kappa B .

Thiol modulation inhibits the <protein>interleukin ( IL ) -1</protein> -mediated activation of an <protein>IL-1 receptor-associated protein kinase</protein> and <protein>NF-kappa B</protein> .


The interleukin-1 receptor type I ( IL-1RI ) is associated with other proteins thus forming a complex system by which IL-1 exerts its various signals .

The <protein>interleukin-1 receptor type I</protein> ( <protein>IL-1RI</protein> ) is associated with other proteins thus forming a <protein>complex system</protein> by which <protein>IL-1</protein> exerts its various signals .


The initiating event is still uncertain , but activation of a recently described receptor-associated protein kinase is one of the earliest events detectable ( Martin et al. , Eur.J.Immunol.1994.24 : 1566 ) .

The initiating event is still uncertain , but activation of a recently described <protein>receptor-associated protein kinase</protein> is one of the earliest events detectable ( Martin et al. , Eur.J.Immunol.1994.24 : 1566 ) .


IL-1 signaling is commonly accompanied by oxidative processes and is thought to be subject to redox regulation .

<protein>IL-1</protein> signaling is commonly accompanied by oxidative processes and is thought to be subject to redox regulation .


We therefore investigated whether the activation of the IL-1RI-associated protein kinase could be a target for redox regulation and whether an altered activity of the kinase could influence IL-1 -mediated NF-kappa B activation .

We therefore investigated whether the activation of the <protein>IL-1RI-associated protein kinase</protein> could be a target for redox regulation and whether an altered activity of the kinase could influence <protein>IL-1</protein> -mediated <protein>NF-kappa B</protein> activation .


A murine T cell line , EL4 , was stimulated with IL-1 with and without pretreatment with different compounds known to influence the cellular redox status .

A <cell_line>murine T cell line</cell_line> , <cell_line>EL4</cell_line> , was stimulated with <protein>IL-1</protein> with and without pretreatment with different compounds known to influence the cellular redox status .


Thiol modifying agents like diamide , menadione , pyrrolidine dithiocarbamate ( PDTC ) , diethyl dithiocarbamate or phenylarsine oxide inhibited the IL-1 -induced activation of the IL-1RI-associated protein kinase .

Thiol modifying agents like diamide , menadione , pyrrolidine dithiocarbamate ( PDTC ) , diethyl dithiocarbamate or phenylarsine oxide inhibited the <protein>IL-1</protein> -induced activation of the <protein>IL-1RI-associated protein kinase</protein> .


N-Acetylcysteine , alpha , alpha'-dipyridyl , aminotriazole or nitrofurantoin did not show any effect .


The inhibition by PDTC was reversible unless glutathione synthesis was blocked by buthionine sulfoximine .


The described conditions which inhibited or prevented the activation of the IL-1RI-associated kinase similarly impaired the activation of NF-kappa B in EL4 cells .

The described conditions which inhibited or prevented the activation of the <protein>IL-1RI-associated kinase</protein> similarly impaired the activation of <protein>NF-kappa B</protein> in <cell_line>EL4 cells</cell_line> .


From these observations we conclude that free thiols in the IL-1RI complex are essential for the activation of the IL-1RI-associated protein kinase and that this process is mandatory for IL-1 signaling leading to NF-kappa B activation .

From these observations we conclude that free thiols in the <protein>IL-1RI complex</protein> are essential for the activation of the <protein>IL-1RI-associated protein kinase</protein> and that this process is mandatory for <protein>IL-1</protein> signaling leading to <protein>NF-kappa B</protein> activation .


Switching gears during T-cell maturation : RANTES and late transcription .

Switching gears during T-cell maturation : <protein>RANTES</protein> and late transcription .


Although much is understood about the induction of genes expressed early ( within 24 h ) after T-cell activation , little is known about the regulation of expression of genes expressed 'late ' ( three or more days ) post-stimulation .

Although much is understood about the induction of <dna>genes</dna> expressed early ( within 24 h ) after T-cell activation , little is known about the regulation of expression of <dna>genes</dna> expressed 'late ' ( three or more days ) post-stimulation .


A better understanding of transcriptional regulation at this important stage of T-cell maturation may yield new insights into T-cell development and new immunotherapeutic targets .


Involvement of different transduction pathways in NF-kappa B activation by several inducers .

Involvement of different transduction pathways in <protein>NF-kappa B</protein> activation by several <dna>inducers</dna> .


Double-stimulation was used to demonstrate that , in a T lymphocytic cell line ( CEM ) , phorbol myristate acetate ( PMA ) rapidly induced NF-kappa B through a signaling pathway which did not involve reactive oxygen species ( ROS ) and was different from the activation triggered by either H2O2 or tumor necrosis factor-alpha ( TNF-alpha ) .

Double-stimulation was used to demonstrate that , in a <cell_line>T lymphocytic cell line</cell_line> ( <cell_line>CEM</cell_line> ) , phorbol myristate acetate ( PMA ) rapidly induced <protein>NF-kappa B</protein> through a signaling pathway which did not involve reactive oxygen species ( ROS ) and was different from the activation triggered by either H2O2 or <protein>tumor necrosis factor-alpha</protein> ( <protein>TNF-alpha</protein> ) .


Since these latter compounds were known to activate NF-kappa B translocation in a redox-sensitive way , we have demonstrated that NF-kappa B activation by PMA was resistant to antioxidant N-acetyl-L-cysteine ( NAC ) and sensitive to kinase inhibitors staurosporine and H7 while activation by H2O2 or TNF-alpha were not .

Since these latter compounds were known to activate <protein>NF-kappa B</protein> translocation in a redox-sensitive way , we have demonstrated that <protein>NF-kappa B</protein> activation by PMA was resistant to antioxidant N-acetyl-L-cysteine ( NAC ) and sensitive to kinase inhibitors staurosporine and H7 while activation by H2O2 or <protein>TNF-alpha</protein> were not .


Blockade of T-cell activation by dithiocarbamates involves novel mechanisms of inhibition of nuclear factor of activated T cells .

Blockade of T-cell activation by dithiocarbamates involves novel mechanisms of inhibition of <protein>nuclear factor of activated T cells</protein> .


Dithiocarbamates ( DTCs ) have recently been reported as powerful inhibitors of NF-kappaB activation in a number of cell types .

Dithiocarbamates ( DTCs ) have recently been reported as powerful inhibitors of <protein>NF-kappaB</protein> activation in a number of cell types .


Given the role of this transcription factor in the regulation of gene expression in the inflammatory response , NF-kappaB inhibitors have been suggested as potential therapeutic drugs for inflammatory diseases .

Given the role of this transcription factor in the regulation of gene expression in the inflammatory response , <protein>NF-kappaB</protein> inhibitors have been suggested as potential therapeutic drugs for inflammatory diseases .


We show here that DTCs inhibited both interleukin 2 ( IL-2 ) synthesis and membrane expression of antigens which are induced during T-cell activation .

We show here that DTCs inhibited both <protein>interleukin 2</protein> ( <protein>IL-2</protein> ) synthesis and membrane expression of <protein>antigens</protein> which are induced during T-cell activation .


This inhibition , which occurred with a parallel activation of c-Jun transactivating functions and expression , was reflected by transfection experiments at the IL-2 promoter level , and involved not only the inhibition of NF-kappaB -driven reporter activation but also that of nuclear factor of activated T cells ( NFAT ) .

This inhibition , which occurred with a parallel activation of <protein>c-Jun</protein> transactivating functions and expression , was reflected by transfection experiments at the <dna>IL-2 promoter</dna> level , and involved not only the inhibition of <protein>NF-kappaB</protein> -driven reporter activation but also that of <protein>nuclear factor of activated T cells</protein> ( <protein>NFAT</protein> ) .


Accordingly , electrophoretic mobility shift assays ( EMSAs ) indicated that pyrrolidine DTC ( PDTC ) prevented NF-kappaB , and NFAT DNA-binding activity in T cells stimulated with either phorbol myristate acetate plus ionophore or antibodies against the CD3-T-cell receptor complex and simultaneously activated the binding of AP-1 .

Accordingly , electrophoretic mobility shift assays ( EMSAs ) indicated that pyrrolidine DTC ( PDTC ) prevented <protein>NF-kappaB</protein> , and <protein>NFAT</protein> DNA-binding activity in <cell_type>T cells</cell_type> stimulated with either phorbol myristate acetate plus ionophore or antibodies against the <protein>CD3-T-cell receptor complex</protein> and simultaneously activated the binding of <protein>AP-1</protein> .


Furthermore , PDTC differentially targeted both NFATp and NFATc family members , inhibiting the transactivation functions of NFATp and mRNA induction of NFATc .

Furthermore , PDTC differentially targeted both <protein>NFATp and NFATc family members</protein> , inhibiting the transactivation functions of <protein>NFATp</protein> and mRNA induction of <protein>NFATc</protein> .


Strikingly , Western blotting and immunocytochemical experiments indicated that PDTC promoted a transient and rapid shuttling of NFATp and NFATc , leading to their accelerated export from the nucleus of activated T cells .

Strikingly , Western blotting and immunocytochemical experiments indicated that PDTC promoted a transient and rapid shuttling of <protein>NFATp</protein> and <protein>NFATc</protein> , leading to their accelerated export from the nucleus of <cell_type>activated T cells</cell_type> .


We propose that the activation of an NFAT kinase by PDTC could be responsible for the rapid shuttling of the NFAT , therefore transiently converting the sustained transactivation of this transcription factor that occurs during lymphocyte activation , and show that c-Jun NH2-terminal kinase ( JNK ) can act by directly phosphorylating NFATp .

We propose that the activation of an NFAT kinase by PDTC could be responsible for the rapid shuttling of the <protein>NFAT</protein> , therefore transiently converting the sustained transactivation of this <protein>transcription factor</protein> that occurs during lymphocyte activation , and show that <protein>c-Jun NH2-terminal kinase</protein> ( <protein>JNK</protein> ) can act by directly phosphorylating <protein>NFATp</protein> .


In addition , the combined inhibitory effects on NFAT and NF-KB support a potential use of DTCs as immunosuppressants .

In addition , the combined inhibitory effects on <protein>NFAT</protein> and <protein>NF-KB</protein> support a potential use of DTCs as immunosuppressants .


Activation of a novel gene in 3q21 and identification of intergenic fusion transcripts with ecotropic viral insertion site I in leukemia .

Activation of a <dna>novel gene</dna> in <dna>3q21</dna> and identification of intergenic fusion transcripts with <dna>ecotropic viral insertion site I</dna> in leukemia .


We have identified a novel gene , GR6 , located within the leukemia breakpoint region of 3q21 , that is normally expressed in early fetal development but not in adult peripheral blood .

We have identified a <dna>novel gene</dna> , <dna>GR6</dna> , located within the leukemia breakpoint region of <dna>3q21</dna> , that is normally expressed in early fetal development but not in adult peripheral blood .


GR6 is activated in the UCSD-AML1 cell line and in a leukemic sample , both of which carry a t ( 3 ; 3 ) ( q21 ; q26 ) .

<dna>GR6</dna> is activated in the <cell_line>UCSD-AML1 cell line</cell_line> and in a <cell_line>leukemic sample</cell_line> , both of which carry a <dna>t ( 3 ; 3 ) ( q21 ; q26 )</dna> .


In UCSD-AML1 , we have also identified fusion transcripts between the ecotropic viral insertion site I ( EVI1 ) gene in 3q26 and GR6 and between EVI1 and Ribophorin I that maps 30 kb telomeric to GR6 in 3q21 .

In <cell_line>UCSD-AML1</cell_line> , we have also identified <rna>fusion transcripts</rna> between the <dna>ecotropic viral insertion site I ( EVI1 ) gene</dna> in <dna>3q26</dna> and <dna>GR6</dna> and between <dna>EVI1</dna> and <dna>Ribophorin I</dna> that maps 30 kb telomeric to <dna>GR6</dna> in <dna>3q21</dna> .


All fusions splice the 5 ' ends of the 3q21 genes into exon 2 of the EVI1 gene , an event that is similar to the normal intergenic splicing of MDS1-EVI1 and to those previously documented in leukemias with t ( 3 ; 21 ) and t ( 3 ; 12 ) , in which acute myelogenous leukemia 1-EVI1 fusions and ETV6-EVI1 fusions , respectively , occur .

All fusions splice the <dna>5 ' ends</dna> of the <dna>3q21 genes</dna> into <dna>exon 2</dna> of the <dna>EVI1</dna> gene , an event that is similar to the normal intergenic splicing of <dna>MDS1-EVI1</dna> and to those previously documented in leukemias with <dna>t ( 3 ; 21 )</dna> and <dna>t ( 3 ; 12 )</dna> , in which acute <dna>myelogenous leukemia 1-EVI1 fusions</dna> and <dna>ETV6-EVI1 fusions</dna> , respectively , occur .


The Ribophorin I-EVI1 fusion in particular may be a common occurrence in t ( 3 ; 3 ) .

The <dna>Ribophorin I-EVI1 fusion</dna> in particular may be a common occurrence in <dna>t ( 3 ; 3 )</dna> .


The A-myb transcription factor in neoplastic and normal B cells .

The <protein>A-myb transcription factor</protein> in <cell_type>neoplastic and normal B cells</cell_type> .


The myb family of transcription factors has been strongly implicated in the regulation of cell growth and differentiation in the haematopoietic system .

The <protein>myb family</protein> of <protein>transcription factors</protein> has been strongly implicated in the regulation of cell growth and differentiation in the haematopoietic system .


The v-myb oncogene , carried by avian defective retroviruses , causes leukaemias in the chicken and transforms haematopoietic cells in vitro .

The <dna>v-myb oncogene</dna> , carried by avian defective retroviruses , causes leukaemias in the chicken and transforms <cell_type>haematopoietic cells</cell_type> in vitro .


Its normal cellular equivalent c-myb , has been shown to promote the proliferation and block the differentiation of haematopoietic cells in several experimental models and is required for fetal haematopoiesis .

Its normal cellular equivalent <dna>c-myb</dna> , has been shown to promote the proliferation and block the differentiation of <cell_type>haematopoietic cells</cell_type> in several experimental models and is required for fetal haematopoiesis .


Two other members of the family have been cloned more recently , A-myb and B-myb , which show sequence homology with c-myb in several domains , of which the DNA binding domain as well as other regulatory domains .

Two other members of the family have been cloned more recently , <protein>A-myb</protein> and <protein>B-myb</protein> , which show sequence homology with <dna>c-myb</dna> in several domains , of which the <protein>DNA binding domain</protein> as well as other <protein>regulatory domains</protein> .


Both have been shown to be transcription factors .

Both have been shown to be <protein>transcription factors</protein> .


B-myb is also involved in the control of proliferation and differentiation , but , unlike c-myb , it is expressed in many cell types .

<protein>B-myb</protein> is also involved in the control of proliferation and differentiation , but , unlike <dna>c-myb</dna> , it is expressed in many cell types .


The third member of the family , A-myb , shows the most restricted pattern of expression , suggesting a very specific role for this transcription factor .

The third member of the <protein>family</protein> , <protein>A-myb</protein> , shows the most restricted pattern of expression , suggesting a very specific role for this <protein>transcription factor</protein> .


A-myb is expressed in a subpopulation of normal B lymphocytes activated in vivo and localised in the germinal center of peripheral lymphoid organs and is not detected at significant levels in all other mature or immature haematopoietic populations studied , including bone marrow cells , T lymphocytes , granulocytes , monocytes , either at rest or after in vitro activation .

<protein>A-myb</protein> is expressed in a subpopulation of <cell_type>normal B lymphocytes</cell_type> activated in vivo and localised in the germinal center of peripheral lymphoid organs and is not detected at significant levels in all other mature or <cell_type>immature haematopoietic populations</cell_type> studied , including <cell_type>bone marrow cells</cell_type> , <cell_type>T lymphocytes</cell_type> , <cell_type>granulocytes</cell_type> , <cell_type>monocytes</cell_type> , either at rest or after in vitro activation .


These studies indicate that A-myb plays a role during a narrow window of normal B cell differentiation .

These studies indicate that <protein>A-myb</protein> plays a role during a narrow window of <cell_type>normal B cell</cell_type> differentiation .


A-myb expression has also been studied in a wide range of neoplastic B cells , representing the whole spectrum of B cell differentiation .

<protein>A-myb</protein> expression has also been studied in a wide range of <cell_type>neoplastic B cells</cell_type> , representing the whole spectrum of B cell differentiation .


A-myb is strongly expressed in Burkitt 's lymphomas ( BL ) and slg+ B-acute lymphoblastic leukaemias ( B-ALL ) and not in all other leukaemias/lymphomas tested , with the exception of a subset of CLL ( about 25 % of cases ) .

<protein>A-myb</protein> is strongly expressed in Burkitt 's lymphomas ( BL ) and slg+ B-acute lymphoblastic leukaemias ( B-ALL ) and not in all other leukaemias/lymphomas tested , with the exception of a subset of CLL ( about 25 % of cases ) .


It is intriguing that the A-myb genome has been localised relatively close to the c-myc gene on chromosome 8 , suggesting that the c-myc translocation in BL and B-ALL may affect A-myb transcription .

It is intriguing that the <dna>A-myb genome</dna> has been localised relatively close to the <dna>c-myc gene</dna> on <dna>chromosome 8</dna> , suggesting that the c-myc translocation in BL and B-ALL may affect <protein>A-myb</protein> transcription .


Studies are in progress to investigate the functional relationship between A-myb and c-myc , particularly in the context of BL cells and to determine whether A-myb is deregulated in these cells .

Studies are in progress to investigate the functional relationship between <protein>A-myb</protein> and <dna>c-myc</dna> , particularly in the context of <cell_type>BL cells</cell_type> and to determine whether <protein>A-myb</protein> is deregulated in these cells .


Expression of bcl-6 protein in normal skin and epidermal neoplasms .

Expression of <protein>bcl-6 protein</protein> in normal skin and epidermal neoplasms .


Bcl-6 protein is a recently identified novel transcription factor whose deregulated expression is associated with diffuse large B cell lymphomas .

<protein>Bcl-6 protein</protein> is a recently identified novel <protein>transcription factor</protein> whose deregulated expression is associated with diffuse large B cell lymphomas .


It was recently shown by us that the protein is located in germinal center B cells and their neoplastic counterparts .

It was recently shown by us that the protein is located in <cell_type>germinal center B cells</cell_type> and their <cell_type>neoplastic counterparts</cell_type> .


In the present study , the expression of bcl-6 protein on normal epidermis , benign , and malignant tumors originating from epidermal cells , and squamous cell carcinoma ( SCC ) cell lines are investigated .

In the present study , the expression of <protein>bcl-6 protein</protein> on normal epidermis , benign , and malignant tumors originating from <cell_type>epidermal cells</cell_type> , and <cell_line>squamous cell carcinoma ( SCC ) cell lines</cell_line> are investigated .


With the use of immunohistochemistry , bcl-6 protein was shown to stain intensely on normal prickle cells , but none to only slightly on epidermal basal cells .

With the use of immunohistochemistry , <protein>bcl-6 protein</protein> was shown to stain intensely on <cell_line>normal prickle cells</cell_line> , but none to only slightly on <cell_type>epidermal basal cells</cell_type> .


Papillomas and keratoacanthomas copied their normal counterparts in the mode of expression .


Various levels of expression were found on seborrheic keratoses , while the expression level on basal cell epitheliomas was low .


Peculiarly , eccrine poromas and undifferentiated spindle-shaped basal cell epitheliomas were totally unstained .


Squamous cell carcinomas showed a variety of expression levels , while two undifferentiated spindle-shaped carcinomas and one undifferentiated SCC cell line remained unstained .

Squamous cell carcinomas showed a variety of expression levels , while two undifferentiated spindle-shaped carcinomas and one undifferentiated <cell_line>SCC cell line</cell_line> remained unstained .


These results suggest that the expression of bcl-6 protein may be associated with morphological differentiation in normal and neoplastic epidermal cells .

These results suggest that the expression of <protein>bcl-6 protein</protein> may be associated with morphological differentiation in normal and <cell_type>neoplastic epidermal cells</cell_type> .


Sp3 mediates transcriptional activation of the leukocyte integrin genes CD11C and CD11B and cooperates with c-Jun to activate CD11C .

<protein>Sp3</protein> mediates transcriptional activation of the <dna>leukocyte integrin genes</dna> <dna>CD11C</dna> and <dna>CD11B</dna> and cooperates with c-Jun to activate <dna>CD11C</dna> .


The leukocyte integrin genes CD11c and CD11b are expressed predominately in myelomonocytic cells .

The <dna>leukocyte integrin genes</dna> <dna>CD11c</dna> and <dna>CD11b</dna> are expressed predominately in myelomonocytic cells .


In previous experiments , the -70 to -65 and -121 to -103 regions of the CD11c promoter and the -66 to -59 region of the CD11b promoter were shown to be essential for Sp1- mediated activation of these genes .

In previous experiments , the <dna>-70 to -65 and -121 to -103 regions</dna> of the <dna>CD11c promoter</dna> and the <dna>-66 to -59 region</dna> of the <dna>CD11b promoter</dna> were shown to be essential for <protein>Sp1-</protein> mediated activation of these genes .


In vivo genomic footprinting had also revealed cell-specific binding of protein , presumably Sp1 , to these regions .

In vivo genomic footprinting had also revealed cell-specific binding of protein , presumably <protein>Sp1</protein> , to these regions .


In this study , electrophoretic mobility shift analysis showed that the Sp1-related factor , Sp3 , also binds at or near these same regions .

In this study , electrophoretic mobility shift analysis showed that the <protein>Sp1-related factor</protein> , <protein>Sp3</protein> , also binds at or near these same regions .


Cotransfection of Sp3 along with CD11c promoter-luciferase constructs into Sp-deficient Drosophila Schneider 2 cells showed that Sp3 could activate the CD11c promoter .

Cotransfection of <protein>Sp3</protein> along with <dna>CD11c promoter-luciferase constructs</dna> into <cell_line>Sp-deficient Drosophila Schneider 2 cells</cell_line> showed that <protein>Sp3</protein> could activate the <dna>CD11c promoter</dna> .


Deletion of both the -70 to -65 and -121 to -103 regions of the CD11c promoter resulted in the loss of activation by Sp3 .

Deletion of both the <dna>-70 to -65 and -121 to -103 regions</dna> of the <dna>CD11c promoter</dna> resulted in the loss of activation by <protein>Sp3</protein> .


Both sites showed activation by Sp3 ; however , the -70 to -65 region was more responsive to Sp3 than to Sp1 .

Both sites showed activation by <protein>Sp3</protein> ; however , the <dna>-70 to -65 region</dna> was more responsive to <protein>Sp3</protein> than to <protein>Sp1</protein> .


Similar transfection analysis of the -66 to -59 region of the CD11b promoter showed Sp3 -dependent expression .

Similar transfection analysis of the <dna>-66 to -59 region</dna> of the <dna>CD11b promoter</dna> showed <protein>Sp3</protein> -dependent expression .


Further , cotransfection analysis in Drosophila cells showed that Sp3 , as was previously shown for Sp1 , also synergizes with c-Jun to activate CD11c .

Further , cotransfection analysis in <cell_line>Drosophila cells</cell_line> showed that <protein>Sp3</protein> , as was previously shown for <protein>Sp1</protein> , also synergizes with <protein>c-Jun</protein> to activate <dna>CD11c</dna> .


Antisense experiments that knocked out endogenous Sp3 expression in the myelomocytic cell line , HL60 , revealed that Sp3 participates in activation of the CD11c and CD11b promoters in vivo .

Antisense experiments that knocked out endogenous <protein>Sp3</protein> expression in the <cell_line>myelomocytic cell line</cell_line> , <cell_line>HL60</cell_line> , revealed that <protein>Sp3</protein> participates in activation of the <dna>CD11c and CD11b promoters</dna> in vivo .


The winged-helix transcription factor Trident is expressed in actively dividing lymphocytes .

The <protein>winged-helix transcription factor</protein> <protein>Trident</protein> is expressed in actively <cell_type>dividing lymphocytes</cell_type> .


We recently identified the winged-helix transcription factor Trident and described its expression pattern in synchronized fibroblasts .

We recently identified the <protein>winged-helix transcription factor</protein> <protein>Trident</protein> and described its expression pattern in <cell_type>synchronized fibroblasts</cell_type> .


We have now studied Trident expression in cell lines , differentiating thymocytes and in lymphocytes derived from peripheral blood .

We have now studied <protein>Trident</protein> expression in <cell_line>cell lines</cell_line> , <cell_type>differentiating thymocytes</cell_type> and in lymphocytes derived from peripheral blood .


During T cell differentiation , expression peaked in the actively dividing immature single positive cells .

During T cell differentiation , expression peaked in the <cell_type>actively dividing immature single positive cells</cell_type> .


In peripheral blood lymphocytes , expression of Trident mRNA was absent , but could be induced upon stimulation with mitogens in vitro .

In peripheral blood lymphocytes , expression of <rna>Trident mRNA</rna> was absent , but could be induced upon stimulation with mitogens in vitro .


These observations imply a function for Trident in dividing lymphocytes .

These observations imply a function for <protein>Trident</protein> in <cell_type>dividing lymphocytes</cell_type> .


Estrogen receptor diminishes DNA-binding activities of chicken GATA-1 and CACCC-binding proteins .

<protein>Estrogen receptor</protein> diminishes DNA-binding activities of <protein>chicken GATA-1</protein> and <protein>CACCC-binding proteins</protein> .


The estrogen receptor ( ER ) repressed erythroid differentiation and erythroid-specific gene expression .

The <protein>estrogen receptor</protein> ( <protein>ER</protein> ) repressed erythroid differentiation and <dna>erythroid-specific gene</dna> expression .


In this study , we investigated the effect of ER alpha ( referred to throughout as ER ) on DNA-binding activities of transcription factors involved in regulating the expression of erythroid-specific genes , and , in particular , the histone H5 gene .

In this study , we investigated the effect of <protein>ER alpha</protein> ( referred to throughout as <protein>ER</protein> ) on DNA-binding activities of <protein>transcription factors</protein> involved in regulating the expression of <dna>erythroid-specific genes</dna> , and , in particular , the <dna>histone H5 gene</dna> .


Using electrophoretic mobility shift assays , we found that in the presence of rabbit reticulocyte lysate , human ER reduced the binding activities of chicken immature erythrocyte nuclear extracted proteins to GATA and CACCC sites in the H5 promoter and enhancer .

Using electrophoretic mobility shift assays , we found that in the presence of rabbit reticulocyte lysate , <protein>human ER</protein> reduced the binding activities of <protein>chicken immature erythrocyte nuclear extracted proteins</protein> to <dna>GATA and CACCC sites</dna> in the <dna>H5 promoter and enhancer</dna> .


In contrast , the binding activities of NF1 and Sp1 were not affected by ER .

In contrast , the binding activities of <protein>NF1</protein> and <protein>Sp1</protein> were not affected by <protein>ER</protein> .


Binding of ER to an estrogen response element was enhanced by addition of rabbit reticulocyte lysate .

Binding of <protein>ER</protein> to an <dna>estrogen response element</dna> was enhanced by addition of rabbit reticulocyte lysate .


This lysate was also necessary for ER to diminish the DNA-binding activity of GATA-1 .

This lysate was also necessary for <protein>ER</protein> to diminish the DNA-binding activity of <protein>GATA-1</protein> .


These results suggest that additional factor ( s ) are necessary for full ER function .

These results suggest that additional factor ( s ) are necessary for full <protein>ER</protein> function .


Both GATA-1 and CACCC-binding proteins are critical for the developmentally regulated expression of erythroid-specific genes .

Both <protein>GATA-1</protein> and <protein>CACCC-binding proteins</protein> are critical for the developmentally regulated expression of <dna>erythroid-specific genes</dna> .


We hypothesize that interference in DNA-binding activities of GATA-1 and CACCC-binding proteins is the mechanism by which the ER inhibits regulation of these genes .

We hypothesize that interference in DNA-binding activities of <protein>GATA-1</protein> and <protein>CACCC-binding proteins</protein> is the mechanism by which the <protein>ER</protein> inhibits regulation of these genes .


Regulation of Id3 cell cycle function by Cdk-2-dependent phosphorylation .

Regulation of <protein>Id3</protein> cell cycle function by Cdk-2-dependent phosphorylation .


The functions of basic helix-loop-helix ( bHLH ) transcription factors in activating differentiation-linked gene expression and in inducing G1 cell cycle arrest are negatively regulated by members of the Id family of HLH proteins .

The functions of <protein>basic helix-loop-helix ( bHLH ) transcription factors</protein> in activating differentiation-linked gene expression and in inducing G1 cell cycle arrest are negatively regulated by members of the <protein>Id family of HLH proteins</protein> .


These bHLH antagonists are induced during a mitogenic signalling response , and they function by sequestering their bHLH targets in inactive heterodimers that are unable to bind to specific gene regulatory ( E box ) sequences .

These <protein>bHLH antagonists</protein> are induced during a mitogenic signalling response , and they function by sequestering their <protein>bHLH targets</protein> in <protein>inactive heterodimers</protein> that are unable to bind to specific <dna>gene regulatory ( E box ) sequences</dna> .


Recently , cyclin E-Cdk2- and cyclin A-Cdk2-dependent phosphorylation of a single conserved serine residue ( Ser5 ) in Id2 has been shown to occur during late G1-to-S phase transition of the cell cycle , and this neutralizes the function of Id2 in abrogating E-box-dependent bHLH homo -or heterodimer complex formation in vitro ( E.Hara , M.Hall , and G.Peters , EMBO J.16 : 332-342 , 1997 ) .

Recently , cyclin E-Cdk2- and cyclin A-Cdk2-dependent phosphorylation of a single conserved serine residue ( Ser5 ) in <protein>Id2</protein> has been shown to occur during late G1-to-S phase transition of the cell cycle , and this neutralizes the function of <protein>Id2</protein> in abrogating <protein>E-box-dependent bHLH</protein> homo -or heterodimer complex formation in vitro ( E.Hara , M.Hall , and G.Peters , EMBO J.16 : 332-342 , 1997 ) .


We now show that an analogous cell-cycle-regulated phosphorylation of Id3 alters the specificity of Id3 for abrogating both E-box-dependent bHLH homo- or heterodimer complex formation in vitro and E-box-dependent reporter gene function in vivo .

We now show that an analogous cell-cycle-regulated phosphorylation of <protein>Id3</protein> alters the specificity of <protein>Id3</protein> for abrogating both <protein>E-box-dependent bHLH</protein> homo- or heterodimer complex formation in vitro and E-box-dependent <dna>reporter gene</dna> function in vivo .


Furthermore , compared with wild-type Id3 , an Id3 Asp5 mutant ( mimicking phosphorylation ) is unable to promote cell cycle S phase entry in transfected fibroblasts , whereas an Id3 Ala5 mutant ( ablating phosphorylation ) displays an activity significantly greater than that of wild-type Id3 protein .

Furthermore , compared with <protein>wild-type Id3</protein> , an <protein>Id3 Asp5 mutant</protein> ( mimicking phosphorylation ) is unable to promote cell cycle S phase entry in <cell_line>transfected fibroblasts</cell_line> , whereas an <protein>Id3 Ala5 mutant</protein> ( ablating phosphorylation ) displays an activity significantly greater than that of <protein>wild-type Id3 protein</protein> .


Cdk2-dependent phosphorylation therefore provides a switch during late G1-to-S phase that both nullifies an early G1 cell cycle regulatory function of Id3 and modulates its target bHLH specificity .

Cdk2-dependent phosphorylation therefore provides a switch during late G1-to-S phase that both nullifies an early G1 cell cycle regulatory function of <protein>Id3</protein> and modulates its target <protein>bHLH</protein> specificity .


These data also demonstrate that the ability of Id3 to promote cell cycle S phase entry is not simply a function of its ability to modulate bHLH heterodimer-dependent gene expression and establish a biologically important mechanism through which Cdk2 and Id-bHLH functions are integrated in the coordination of cell proliferation and differentiation .

These data also demonstrate that the ability of <protein>Id3</protein> to promote cell cycle S phase entry is not simply a function of its ability to modulate <protein>bHLH</protein> heterodimer-dependent gene expression and establish a biologically important mechanism through which Cdk2 and Id-bHLH functions are integrated in the coordination of cell proliferation and differentiation .


Molecular cloning and functional characterization of murine cDNA encoding transcription factor NFATc .

Molecular cloning and functional characterization of <protein>murine cDNA encoding transcription factor</protein> <protein>NFATc</protein> .


Transcription factors of the NFAT ( nuclear factor of activated T cells ) family play important roles in immune and inflammatory responses by regulating the expression of genes encoding cytokines and immunoregulatory proteins .

<protein>Transcription factors</protein> of the <protein>NFAT ( nuclear factor of activated T cells ) family</protein> play important roles in immune and inflammatory responses by regulating the expression of genes encoding <protein>cytokines</protein> and <protein>immunoregulatory proteins</protein> .


Here we describe cloning and characterization of full-length cDNA encoding murine ( m ) NFATc which predicts that the protein has all the conserved structural motifs of NFAT family members , including the rel homology domain , the NFAT homology domain and the nuclear translocation signals .

Here we describe cloning and characterization of full-length <protein>cDNA encoding murine ( m ) NFATc</protein> which predicts that the protein has all the <protein>conserved structural motifs</protein> of <protein>NFAT family members</protein> , including the rel homology domain , the <protein>NFAT homology domain</protein> and the nuclear translocation signals .


mNFATc complexed with AP-1 bound specifically to the murine IL-2 NFAT recognition sequence and activated transcription from the co-transfected IL-2 promoter in COS-7 cells .

<protein>mNFATc</protein> complexed with <protein>AP-1</protein> bound specifically to the <protein>murine IL-2 NFAT recognition sequence</protein> and activated transcription from the <dna>co-transfected IL-2 promoter</dna> in <cell_line>COS-7 cells</cell_line> .


Northern blot analysis showed that the cDNA probe hybridized with a 4.5 kb transcript which is highly inducible in murine T cells .

Northern blot analysis showed that the <dna>cDNA probe</dna> hybridized with a 4.5 kb transcript which is highly inducible in <cell_type>murine T cells</cell_type> .


By Northern and in situ hybridization , mNFATc transcript was detected from the early stage of development .

By Northern and in situ hybridization , <rna>mNFATc transcript</rna> was detected from the early stage of development .


In the mouse embryo , mNFATc transcript was strongly expressed in thymus , lung and submandibular gland and weakly in skeletal muscle and heart suggesting that mNFATc may have a role both in embryogenesis and in mature T cells .

In the mouse embryo , <rna>mNFATc transcript</rna> was strongly expressed in thymus , lung and submandibular gland and weakly in skeletal muscle and heart suggesting that <protein>mNFATc</protein> may have a role both in embryogenesis and in <cell_type>mature T cells</cell_type> .


TNFalpha cooperates with the protein kinase A pathway to synergistically increase HIV-1 LTR transcription via downstream TRE-like cAMP response elements .

<protein>TNFalpha</protein> cooperates with the <protein>protein kinase A</protein> pathway to synergistically increase <dna>HIV-1 LTR</dna> transcription via downstream <dna>TRE-like cAMP response elements</dna> .


Activating protein-1 ( AP-1 ) binding TPA responsive elements ( TRE ) are located downstream of the transcription initiation site in the U5 region of the HIV-1 long terminal repeat ( LTR ) .

<protein>Activating protein-1</protein> ( <protein>AP-1</protein> ) binding <dna>TPA responsive elements</dna> ( <dna>TRE</dna> ) are located downstream of the <dna>transcription initiation site</dna> in the <dna>U5 region</dna> of the <dna>HIV-1 long terminal repeat</dna> ( <dna>LTR</dna> ) .


These downstream sequence elements , termed DSE , can bind both AP-1 and CREB/ATF transcription factors .

These <dna>downstream sequence elements</dna> , termed <dna>DSE</dna> , can bind both <protein>AP-1</protein> and <protein>CREB/ATF</protein> <protein>transcription factors</protein> .


Recently , we demonstrated that the DSE are also cAMP-responsive elements ( CRE ) , since they mediated activation signals elicited by cholera toxin ( Ctx ) , a potent activator of the cAMP-dependent protein kinase A ( PKA ) signal transduction pathway .

Recently , we demonstrated that the <dna>DSE</dna> are also <dna>cAMP-responsive elements</dna> ( <dna>CRE</dna> ) , since they mediated activation signals elicited by <protein>cholera toxin</protein> ( <protein>Ctx</protein> ) , a potent activator of the cAMP-dependent <protein>protein kinase A</protein> ( <protein>PKA</protein> ) signal transduction pathway .


In the present study , we demonstrate that the HIV-1 DSE can mediate the transcriptional synergy elicited by the combination of Ctx and TNFalpha .

In the present study , we demonstrate that the <dna>HIV-1 DSE</dna> can mediate the transcriptional synergy elicited by the combination of <protein>Ctx</protein> and <protein>TNFalpha</protein> .


Ctx combined with TNFalpha or IL-1beta to produce a synergistic increase in p24 antigen production in U1 promonocytic cells .

<protein>Ctx</protein> combined with <protein>TNFalpha</protein> or <protein>IL-1beta</protein> to produce a synergistic increase in <protein>p24 antigen</protein> production in <cell_line>U1 promonocytic cells</cell_line> .


Transfection studies of LTR reporter constructs indicated that mutation of the DSE sites abrogated the LTR -mediated synergy induced by Ctx and TNFalpha , whereas the synergy induced by Ctx and IL-1beta was unaffected , suggesting TNFalpha and IL-1beta cooperate differently with the cAMP /PKA activation pathway to induce HIV-1 expression in U1 cells .

Transfection studies of <dna>LTR reporter constructs</dna> indicated that mutation of the <dna>DSE sites</dna> abrogated the <dna>LTR</dna> -mediated synergy induced by <protein>Ctx</protein> and <protein>TNFalpha</protein> , whereas the synergy induced by <protein>Ctx</protein> and <protein>IL-1beta</protein> was unaffected , suggesting <protein>TNFalpha</protein> and <protein>IL-1beta</protein> cooperate differently with the cAMP <protein>/PKA</protein> activation pathway to induce HIV-1 expression in <cell_line>U1 cells</cell_line> .


Because the DSE are also TRE sites , we assessed the effect of the agonist combinations on AP-1 -dependent transcription .

Because the <dna>DSE</dna> are also <dna>TRE</dna> sites , we assessed the effect of the agonist combinations on <protein>AP-1</protein> -dependent transcription .


TNFalpha as well as IL-1beta cooperated with Ctx to produce a synergistic activation of AP-1 -mediated transcription .

<protein>TNFalpha</protein> as well as <protein>IL-1beta</protein> cooperated with <protein>Ctx</protein> to produce a synergistic activation of <protein>AP-1</protein> -mediated transcription .


These data indicate that the TRE-like cAMP-responsive DSE sites within the 5'-untranslated leader can mediate the transcriptional cooperativity between TNFalpha and the cAMP /PKA pathway .

These data indicate that the <dna>TRE-like cAMP-responsive DSE sites</dna> within the <dna>5'-untranslated leader</dna> can mediate the transcriptional cooperativity between <protein>TNFalpha</protein> and the cAMP <protein>/PKA</protein> pathway .


Since the DSE and TRE sites can not bind CREB/ATF homodimers , we propose a mechanism in which the HIV-1 DSE bind heterodimers composed of both AP-1 and CREB/ATF proteins .

Since the <dna>DSE</dna> and <dna>TRE</dna> sites can not bind <protein>CREB/ATF homodimers</protein> , we propose a mechanism in which the HIV-1 <dna>DSE</dna> bind <protein>heterodimers</protein> composed of both <protein>AP-1 and CREB/ATF proteins</protein> .


Copyright 1997 Academic Press .


Association of glucocorticoid insensitivity with increased expression of glucocorticoid receptor beta .

Association of glucocorticoid insensitivity with increased expression of <protein>glucocorticoid receptor beta</protein> .


In many chronic inflammatory disorders , glucocorticoid ( GC ) insensitivity is a challenging clinical problem associated with life-threatening disease progression .


The molecular basis of GC insensitivity , however , is unknown .


Alternative splicing of the GC receptor ( R ) pre-messenger RNA generates a second GCR , termed GCR-beta , which does not bind GCs but antagonizes the transactivating activity of the classic GCR , termed GCR-alpha .

Alternative splicing of the <rna>GC receptor ( R ) pre-messenger RNA</rna> generates a second <protein>GCR</protein> , termed <protein>GCR-beta</protein> , which does not bind GCs but antagonizes the transactivating activity of the classic <protein>GCR</protein> , termed <protein>GCR-alpha</protein> .


In the current study , we demonstrate that GC-insensitive asthma is associated with a significantly higher number of GCR-beta-immunoreactive cells in peripheral blood than GC-sensitive asthmatics or normal controls .

In the current study , we demonstrate that GC-insensitive asthma is associated with a significantly higher number of <cell_type>GCR-beta-immunoreactive cells</cell_type> in peripheral blood than GC-sensitive asthmatics or normal controls .


Furthermore , we show that patients with GC-insensitive asthma have cytokine-induced abnormalities in the DNA binding capability of the GCR .

Furthermore , we show that patients with GC-insensitive asthma have cytokine-induced abnormalities in the DNA binding capability of the <protein>GCR</protein> .


These abnormalities can be reproduced by transfection of cell lines with the GCR-beta gene resulting in significant reduction of their GCR-alpha DNA binding capacity .

These abnormalities can be reproduced by transfection of <cell_line>cell lines</cell_line> with the <protein>GCR-beta</protein> gene resulting in significant reduction of their <protein>GCR-alpha</protein> DNA binding capacity .


We conclude that increased expression of GCR-beta is cytokine inducible and may account for GC insensitivity in this common inflammatory condition .

We conclude that increased expression of <protein>GCR-beta</protein> is <protein>cytokine</protein> inducible and may account for GC insensitivity in this common inflammatory condition .


Epstein-Barr virus EBNA3C represses Cp , the major promoter for EBNA expression , but has no effect on the promoter of the cell gene CD21 .

Epstein-Barr virus <protein>EBNA3C</protein> represses <dna>Cp</dna> , the major promoter for <protein>EBNA</protein> expression , but has no effect on the promoter of the <dna>cell gene CD21</dna> .


EBNA3C is a potent repressor of transcription when bound to DNA as a fusion with the DNA binding domain ( DBD ) of GALA .

<protein>EBNA3C</protein> is a potent repressor of transcription when bound to DNA as a fusion with the <protein>DNA binding domain</protein> ( <protein>DBD</protein> ) of GALA .


A survey of promoters has revealed that the wild-type , unfused EBNA3C can specifically repress expression from reporter plasmids containing the Epstein-Barr virus Cp latency-associated promoter .

A survey of promoters has revealed that the <protein>wild-type , unfused EBNA3C</protein> can specifically repress expression from <dna>reporter plasmids</dna> containing the Epstein-Barr virus <dna>Cp latency-associated promoter</dna> .


Repression of Cp activity required amino acids 207 to 368 , which encompasses a region resembling a basic DBD adjacent to a leucine zipper DNA binding motif and a site which binds to the cellular factor CBF1/RBP-Jkappa .

Repression of <dna>Cp</dna> activity required <protein>amino acids 207 to 368</protein> , which encompasses a region resembling a <protein>basic DBD</protein> adjacent to a <protein>leucine zipper DNA binding motif</protein> and a site which binds to the <protein>cellular factor CBF1/RBP-Jkappa</protein> .


However , amino acids 207 to 368 are dispensable when the protein is bound to DNA as a fusion with the GAL4 DBD , thus implicating this region in DNA binding .

However , amino acids 207 to 368 are dispensable when the protein is bound to DNA as a fusion with the GAL4 <protein>DBD</protein> , thus implicating this region in DNA binding .


Mutation of the CBF1/RBP-Jkappa binding site in EBNA3C abrogated repression , strongly suggesting that CBF1/RBP-Jkappa is necessary for targeting the viral protein to Cp .

Mutation of the <protein>CBF1/RBP-Jkappa binding site</protein> in <protein>EBNA3C</protein> abrogated repression , strongly suggesting that <protein>CBF1/RBP-Jkappa</protein> is necessary for targeting the viral protein to <dna>Cp</dna> .


Consistent with this result , mutation of the EBNA2 response element ( a CBF1/RBP-Jkappa binding site ) in Cp also prevented significant repression .

Consistent with this result , mutation of the <dna>EBNA2 response element</dna> ( a <dna>CBF1/RBP-Jkappa binding site</dna> ) in <dna>Cp</dna> also prevented significant repression .


In addition , amino acids 346 to 543 , which were previously defined as important for the repressor activity of the GAL4-EBNA3C fusion proteins , also appear to be necessary for the repression of Cp .

In addition , <protein>amino acids 346 to 543</protein> , which were previously defined as important for the repressor activity of the <protein>GAL4-EBNA3C fusion proteins</protein> , also appear to be necessary for the repression of <dna>Cp</dna> .


Since repression by these fusions was not observed in all cell types , it seems likely that EBNA3C either depends on a corepressor which may interact with amino acids 346 to 543 or is modified in a cell-specific manner in order to repress .

Since repression by these fusions was not observed in all cell types , it seems likely that <protein>EBNA3C</protein> either depends on a <protein>corepressor</protein> which may interact with <protein>amino acids 346 to 543</protein> or is modified in a cell-specific manner in order to repress .


These data are consistent with EBNA3C contributing to the regulation of EBNA expression in latently infected B cells through CBF1/RBP-Jkappa and another factor , but this need not directly involve EBNA2 .

These data are consistent with <protein>EBNA3C</protein> contributing to the regulation of <protein>EBNA</protein> expression in <cell_type>latently infected B cells</cell_type> through <protein>CBF1/RBP-Jkappa</protein> and another factor , but this need not directly involve <protein>EBNA2</protein> .


Finally , although it has been reported that EBNA3C can upregulate CD21 in some B cells , we were unable to demonstrate any effect of EBNA3C on reporter plasmids which contain the CD21 promoter .

Finally , although it has been reported that <protein>EBNA3C</protein> can upregulate <protein>CD21</protein> in some <cell_type>B cells</cell_type> , we were unable to demonstrate any effect of <protein>EBNA3C</protein> on <dna>reporter plasmids</dna> which contain the <dna>CD21 promoter</dna> .


Relationship between glucocorticoid receptor and response to glucocorticoid therapy in ulcerative colitis .

Relationship between <protein>glucocorticoid receptor</protein> and response to glucocorticoid therapy in ulcerative colitis .


PURPOSE : To clarify the relationship between the glucocorticoid receptor and the effectiveness of glucocorticoid therapy in patients with ulcerative colitis , we investigated the number and apparent dissociation constant of glucocorticoid receptor in peripheral blood mononuclear leukocytes of patients with ulcerative colitis .

PURPOSE : To clarify the relationship between the <protein>glucocorticoid receptor</protein> and the effectiveness of glucocorticoid therapy in patients with ulcerative colitis , we investigated the number and apparent dissociation constant of <protein>glucocorticoid receptor</protein> in <cell_type>peripheral blood mononuclear leukocytes</cell_type> of patients with ulcerative colitis .


MATERIALS AND METHODS : Eleven patients with ulcerative colitis ( 5 who responded to intravenous glucocorticoids and 6 who did not ) and ten control subjects were studied .


The number and apparent dissociation constant of glucocorticoid receptor were measured using a whole-cell binding assay .

The number and apparent dissociation constant of <protein>glucocorticoid receptor</protein> were measured using a whole-cell binding assay .


Results were expressed as a median ( interquartile range ) .


RESULTS : The number of glucocorticoid receptors from the six nonresponders , five responders , and ten healthy controls were 4922 ( range , 4484-5643 ) , 3413 ( range , 3183-4450 ) , and 3610 ( range , 2594-3979 ) binding sites/cell , respectively .

RESULTS : The number of <protein>glucocorticoid receptors</protein> from the six nonresponders , five responders , and ten healthy controls were 4922 ( range , 4484-5643 ) , 3413 ( range , 3183-4450 ) , and 3610 ( range , 2594-3979 ) binding sites/cell , respectively .


The apparent dissociation constant of the glucocorticoid receptors from the nonresponders , responders , and healthy controls were 7.03 ( range , 5.66-10 ) , 4.27 ( range , 4-5.13 ) , and 6.18 ( range , 5.86-6.74 ) nM , respectively .

The apparent dissociation constant of the <protein>glucocorticoid receptors</protein> from the nonresponders , responders , and healthy controls were 7.03 ( range , 5.66-10 ) , 4.27 ( range , 4-5.13 ) , and 6.18 ( range , 5.86-6.74 ) nM , respectively .


Nonresponders had a significant increase both in the number of binding sites and in the apparent dissociation constant compared with responders ( P = 0.045 ; P = 0.029 ) .

Nonresponders had a significant increase both in the number of <protein>binding sites</protein> and in the apparent dissociation constant compared with responders ( P = 0.045 ; P = 0.029 ) .


CONCLUSIONS : The increased number and apparent dissociation constant of glucocorticoid receptor are closely associated with the effectiveness of glucocorticoid therapy .

CONCLUSIONS : The increased number and apparent dissociation constant of <protein>glucocorticoid receptor</protein> are closely associated with the effectiveness of glucocorticoid therapy .


The measurement of the number and apparent dissociation constant of glucocorticoid receptor may be useful in predicting response to glucocorticoids .

The measurement of the number and apparent dissociation constant of <protein>glucocorticoid receptor</protein> may be useful in predicting response to glucocorticoids .


Cooperation of binding sites for STAT6 and NF kappa B/rel in the IL-4 -induced up-regulation of the human IgE germline promoter .

Cooperation of <protein>binding sites</protein> for <protein>STAT6</protein> and <protein>NF kappa B/rel</protein> in the <protein>IL-4</protein> -induced up-regulation of the <dna>human IgE germline promoter</dna> .


Ig heavy chain class switching is directed by cytokines inducing transcription from unrearranged CH genes .

<protein>Ig heavy chain</protein> class switching is directed by <protein>cytokines</protein> inducing transcription from unrearranged <dna>CH genes</dna> .


Subsequently , such primed cells can undergo switch recombination to express the selected new isotype .

Subsequently , such <cell_type>primed cells</cell_type> can undergo switch recombination to express the selected new isotype .


In the case of IgE class switching , IL-4 activates the IgE germline promoter by inducing the interaction of the transcription factor STAT6 ( IL-4STAT ) with a responsive DNA element in the proximal region of the promoter .

In the case of <protein>IgE</protein> class switching , <protein>IL-4</protein> activates the <dna>IgE germline promoter</dna> by inducing the interaction of the <protein>transcription factor</protein> <protein>STAT6</protein> ( <protein>IL-4STAT</protein> ) with a <dna>responsive DNA element</dna> in the <dna>proximal region</dna> of the <dna>promoter</dna> .


This study describes the characterization of two additional cis-acting elements that interact with members of the NF kappa B/rel transcription factor family in an IL-4 -independent fashion .

This study describes the characterization of two additional <dna>cis-acting elements</dna> that interact with members of the <protein>NF kappa B/rel transcription factor family</protein> in an <protein>IL-4</protein> -independent fashion .


Electrophoretic mobility shift assays show that the nucleoprotein complex formed on the upstream site ( NF kappa B1 ) contains the classical p50/p65 heterodimer .

Electrophoretic mobility shift assays show that the <protein>nucleoprotein complex</protein> formed on the <dna>upstream site</dna> ( <protein>NF kappa B1</protein> ) contains the <protein>classical p50/p65 heterodimer</protein> .


The complex on the proximal site ( NF kappa B2 ) appears to be composed of p50 and relB .

The complex on the <dna>proximal site</dna> ( <protein>NF kappa B2</protein> ) appears to be composed of <protein>p50</protein> and <protein>relB</protein> .