Input:
Lipopolysaccharide induction of the tumor necrosis factor-alpha promoter in human monocytic cells. Regulation by Egr-1, c-Jun, and NF-kappaB transcription factors.
Biosynthesis of tumor necrosis factor-alpha (TNF-alpha) is predominantly by cells of the monocytic lineage. This study examined the role of various cis-acting regulatory elements in the lipopolysaccharide (LPS) induction of the human TNF-alpha promoter in cells of monocytic lineage. Functional analysis of monocytic THP-1 cells transfected with plasmids containing various lengths of TNF-alpha promoter localized enhancer elements in a region (-182 to -37 base pairs (bp)) that were required for optimal transcription of the TNF-alpha gene in response to LPS. Two regions were identified: region I (-182 to -162 bp) contained an overlapping Sp1/Egr-1 site, and region II (-119 to -88) contained CRE and NF-kappaB (designated kappaB3) sites. In unstimulated THP-1, CRE-binding protein and, to a lesser extent, c-Jun complexes were found to bind to the CRE site. LPS stimulation increased the binding of c-Jun-containing complexes. In addition, LPS stimulation induced the binding of cognate nuclear factors to the Egr-1 and kappaB3 sites, which were identified as Egr-1 and p50/p65, respectively. The CRE and kappaB3 sites in region II together conferred strong LPS responsiveness to a heterologous promoter, whereas individually they failed to provide transcriptional activation. Furthermore, increasing the spacing between the CRE and the kappaB3 sites completely abolished LPS induction, suggesting a cooperative interaction between c-Jun complexes and p50/p65. These studies indicate that maximal LPS induction of the TNF-alpha promoter is mediated by concerted participation of at least two separate cis-acting regulatory elements.

promoter

promoter

promoter

tumor necrosis factor-alpha

c-Jun

tumor necrosis factor-alpha

TNF-alpha

TNF-alpha

TNF-alpha

TNF-alpha

c-Jun

c-Jun

c-Jun

TNF-alpha

bind

c-Jun

binding

c-Jun

binding

binding

binding

Biosynthesis

TNF-alpha

induction

tumor necrosis factor-alpha

promoter

induction

TNF-alpha

promoter

required

in response to

in response to

transcription

increased

binding

induction

TNF-alpha

promoter

Regulation

induction

Regulation

induction

c-Jun

role

induction

mediated

induction

transcription

TNF-alpha

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Involvement of Rel, Fos, and Jun proteins in binding activity to the IL-2 promoter CD28 response element/AP-1 sequence in human T cells.
CD28 is an important costimulatory molecule in the activation of human T cells. Costimulation of T cells through both the Ag receptor and CD28 leads to high level IL-2 production, which is vital to the development of an immune response in vivo. Previous reports have suggested the CD28 stimulation contributes to the activation of the IL-2 promoter by up-regulating the activity of several transcription factors, including AP-1 and nuclear factor-kappaB (NF-kappaB)/Rel family members as well as an uncharacterized transcription factor called CD28 response complex. While several lines of investigation have suggested that NF-kappaB/Rel family members make up the CD28 response complex transcription factor, other work has not supported this conclusion. Recent studies suggest that the CD28 response element (CD28RE) does not function independently but works instead in conjunction with the adjacent promoter proximal AP-1-binding site and this hypothesis is confirmed here. Also in the current study, binding activity to the CD28RE/AP-1 sequence of the IL-2 promoter is evaluated. Although four specific complexes can be detected binding to this sequence, only one of these complexes is specific for both the CD28RE and the adjacent AP-1 site. Of the NF-kappaB/Rel family members tested, this CD28RE/AP-1-specific complex contains predominantly c-Rel, despite the fact that both p50 and RelA can efficiently bind to the CD28RE. c-Fos and c-Jun are also found in this CD28RE/AP-1-specific complex. These data indicate that functional complexes encompassing both the CD28RE and the AP-1-binding sites influence IL-2 promoter activity in CD28-costimulated T cells.

promoter

CD28RE/AP-1 sequence

promoter

Fos

Jun

c-Rel

RelA

c-Fos

c-Jun

binding activity

CD28RE/AP-1 sequence

binding

CD28RE/AP-1 sequence

bind

bind

RelA

production

leads

high level

high level

production

contributes

activation

activation

promoter

influence

promoter

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The word or phrase that expresses the event.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
c-Myb and Ets proteins synergize to overcome transcriptional repression by ZEB.
The Zfh family of zinc finger/homeodomain proteins was first identified in Drosophila where it is required for differentiation of tissues such as the central nervous system and muscle. ZEB, a vertebrate homolog of Zfh-1, binds a subset of E boxes and blocks myogenesis through transcriptional repression of muscle genes. We present evidence here that ZEB also has an important role in controlling hematopoietic gene transcription. Two families of transcription factors that are required for normal hematopoiesis are c-Myb and Ets. These factors act synergistically to activate transcription, and this synergy is required for transcription of at least several important hematopoietic genes. ZEB blocks the activity of c-Myb and Ets individually, but together the factors synergize to resist this repression. Such repression imposes a requirement for both c-Myb and Ets for transcriptional activity, providing one explanation for why synergy between these factors is important. The balance between repression by ZEB and transcriptional activation by c-Myb/Ets provides a flexible regulatory mechanism for controlling gene expression in hematopoietic cells. We demonstrate that one target of this positive/negative regulation in vivo is the alpha4 integrin, which play a key role in normal hematopoiesis and function of mature leukocytes.

c-Myb

ZEB

ZEB

ZEB

c-Myb

ZEB

c-Myb

c-Myb

ZEB

c-Myb

alpha4 integrin

binds

ZEB

blocks

ZEB

c-Myb

synergize to resist

blocks

c-Myb

positive/negative regulation

alpha4 integrin

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
GABP factors bind to a distal interleukin 2 (IL-2) enhancer and contribute to c-Raf-mediated increase in IL-2 induction.
Triggering of the T-cell receptor-CD3 complex activates two major signal cascades in T lymphocytes, (i) Ca2+-dependent signal cascades and (ii) protein kinase cascades. Both signal cascades contribute to the induction of the interleukin 2 (IL-2) gene during T-cell activation. Prominent protein kinase cascades are those that activate mitogen-activated protein (MAP) kinases. We show here that c-Raf, which is at the helm of the classic MAP-Erk cascade, contributes to IL-2 induction through a distal enhancer element spanning the nucleotides from positions -502 to -413 in front of the transcriptional start site of the IL-2 gene. Induction of this distal IL-2 enhancer differs from induction of the proximal IL-2 promoter-enhancer, since it is induced by phorbol esters alone and independent from Ca2+ signals. In DNA-protein binding studies, we detected the binding of transcription factors GABP alpha and -beta to a dyad symmetry element (DSE) of the distal enhancer, which is formed by palindromic binding sites of Ets-like factors. Introduction of point mutations suppressing GABP binding to the DSE interfered with the induction of the distal enhancer and the entire IL-2 promoter-enhancer, while overexpression of both GABP factors enhanced the IL-2 promoter-enhancer induction. Overexpression of BXB, a constitutive active version of c-Raf, and of further members of the Ras-Raf-Erk signal cascade exerted an increase of GABP-mediated promoter-enhancer induction. In conjunction with previously published data on c-Raf-induced phosphorylation of GABP factors (E.Flory, A. Hoffmeyer, U.Smola, U.R.Rapp, and J.T.Bruder, J.Virol.70:2260- 2268, 1996), these results indicate a contribution of GABP factors to the Raf-mediated enhancement of IL-2 induction during T-cell activation.

enhancer

promoter-enhancer

promoter-enhancer

promoter-enhancer

interleukin 2

c-Raf

interleukin 2

c-Raf

GABP alpha

-beta

BXB

c-Raf

c-Raf

bind

interleukin 2

enhancer

binding

GABP alpha

binding

-beta

overexpression

GABP alpha

overexpression

-beta

Overexpression

BXB

interfered

induction

increase

c-Raf

induction

induction

induction

induction

induction

promoter-enhancer

induced

promoter-enhancer

independent

promoter-enhancer

induction

promoter-enhancer

overexpression

overexpression

enhanced

overexpression

induction

induction

promoter-enhancer

Overexpression

Overexpression

enhancement

induction

induction

contribute

increase

contribute

induction

contribution

enhancement

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Genomic organization, sequence, and transcriptional regulation of the human eotaxin gene.
Eotaxin is an eosinophil specific beta-chemokine assumed to be involved in eosinophilic inflammatory diseases such as atopic dermatitis, allergic rhinitis, asthma and parasitic infections. Its expression is stimulus- and cell-specific. We here describe the genomic organisation (3 exons of 132, 112 and 542 bp and 2 introns of 1211 and 378 bp) and sequence including 3 kb of DNA from the immediate 5' upstream region of the human eotaxin gene. Among the regulatory promoter elements potentially regulating eotaxin gene expression and/or mediating the effects of anti-inflammatory drugs we identified consensus sequences known to interact with nuclear factors like NF-IL6, AP-1, a NF-kappa-B like consensus sequence and gamma-interferon- as well as glucocorticoid response elements.

eotaxin

Eotaxin

eotaxin

eotaxin

gamma-interferon

interact

expression

Eotaxin

expression

eotaxin

Genomic organization

eotaxin

transcriptional regulation

eotaxin

regulating

expression

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Dysregulation of monocytic nuclear factor-kappa B by oxidized low-density lipoprotein.
Nuclear factor-kappa B (NF-kappa B)/Rel transcription factors may be involved in atherosclerosis, as is suggested by the presence of activated NF-kappa B in human atherosclerotic lesions. The aim of the present study was to investigate the effects of oxidized LDL (oxLDL) on the NF-kappa B system in human THP-1 monocytic cells as well as adherent monocytes. Our results demonstrate that short-term incubation of these cells with oxLDL activated p50/p65 containing NF-kappa B dimers and induced the expression of the target gene IL-8. This activation of NF-kappa B was inhibited by the antioxidant and H2O2 scavenger pyrrolidine dithiocarbamate and the proteasome inhibitor PSI. The oxLDL-induced NF-kappa B activation was accompanied by an initial depletion of I kappa B-alpha followed by a slight transient increase in the level of this inhibitor protein. In contrast, long-term treatment with oxLDL prevented the lipopolysaccharide-induced depletion of I kappa B-alpha, accompanied by an inhibition of both NF-kappa B activation and the expression of tumor necrosis factor-alpha and interleukin-1 beta genes. These observations provide additional evidence that oxLDL is a potent modulator of gene expression and suggest that (dys)regulation of NF-kappa B/Rel is likely to play an important role in atherogenesis.

I kappa B-alpha

I kappa B-alpha

tumor necrosis factor-alpha

interleukin-1 beta

expression

tumor necrosis factor-alpha

expression

interleukin-1 beta

depletion

I kappa B-alpha

prevented

depletion

depletion

I kappa B-alpha

inhibition

expression

increase

I kappa B-alpha

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Suppression of MHC class II expression by human class II trans-activator constructs lacking the N-terminal domain.
The class II trans-activator (CIITA) is a bi- or multi-functional domain protein which plays a critical role in the expression of MHC class II genes. We report that removal of the N-terminal 151 amino acids, encompassing all of the acidic domain but leaving intact the proline/serine/threonine-rich domain, results in a mutant protein with potent suppressive properties for MHC class II expression. HeLa cells stably or transiently transfected with mutant CIITA constructs showed up to 99% suppression of MHC class II antigen induction by IFN-gamma and marked suppression of HLA-DRA mRNA expression. Transient transfection of a B lymphoma line resulted in up to 89% reduction of constitutive MHC class II expression within 5 days and suppression of HLA-DRA mRNA synthesis.

class II trans-activator

CIITA

CIITA

IFN-gamma

transfected

CIITA

transfection

CIITA

transfected

transfected

transfection

transfection

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
CD30-dependent degradation of TRAF2: implications for negative regulation of TRAF signaling and the control of cell survival.
CD30 is a cell-surface receptor that can augment lymphocyte activation and survival through its ability to induce the transcription factor NF-kappaB. CD30, however, has also been implicated in the induction of apoptotic cell death of lymphocytes. Here we show that one of the effects of CD30 signal transduction is to render cells sensitive to apoptosis induced by the type 1 tumor necrosis factor receptor (TNFR1). This sensitization is dependent on the TRAF-binding sites within the CD30 cytoplasmic domain. One of the proteins that binds to these sites is TRAF2, a signal transduction molecule that is also utilized by TNFR1 to mediate the activation of several downstream kinases and transcription factors. During CD30 signal transduction, we found that binding of TRAF2 to the cytoplasmic domain of CD30 results in the rapid depletion of TRAF2 and the associated protein TRAF1 by proteolysis. These data suggest a model in which CD30 limits its own ability to transduce cell survival signals through signal-coupled depletion of TRAF2. Depletion of intracellular TRAF2 and its coassociated proteins also increased the sensitivity of the cell to undergoing apoptosis during activation of death-inducing receptors such as TNFR1. Consistent with this hypothesis, expression of a dominant-negative form of TRAF2 was found to potentiate TNFR1-mediated death. These studies provide a potential mechanism through which CD30, as well as other TRAF-binding members of the TNFR superfamily, can negatively regulate cell survival.

TRAF-binding sites

type 1 tumor necrosis factor receptor

binds

TRAF-binding sites

binding

results

binding

proteolysis

activation

degradation

proteolysis

dependent

degradation

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
TNFalpha cooperates with the protein kinase A pathway to synergistically increase HIV-1 LTR transcription via downstream TRE-like cAMP response elements.
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). These downstream sequence elements, termed DSE, can bind both AP-1 and CREB/ATF transcription factors. 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. In the present study, we demonstrate that the HIV-1 DSE can mediate the transcriptional synergy elicited by the combination of Ctx and TNFalpha. Ctx combined with TNFalpha or IL-1beta to produce a synergistic increase in p24 antigen production in U1 promonocytic cells. 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. Because the DSE are also TRE sites, we assessed the effect of the agonist combinations on AP-1-dependent transcription. TNFalpha as well as IL-1beta cooperated with Ctx to produce a synergistic activation of AP-1-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. Since the DSE and TRE sites cannot 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. Copyright 1997 Academic Press.

TNFalpha

TNFalpha

TNFalpha

p24 antigen

TNFalpha

TNFalpha

TNFalpha

TNFalpha

production

p24 antigen

to produce a synergistic increase

TNFalpha

production

to produce a synergistic increase

production

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
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 Epstein-Barr virus latent membrane protein 1 (LMP1) is essential for the transformation of B lymphocytes into lymphoblastoid cell lines. 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. One transformation effector site (TES1), located within the membrane proximal 45 residues of the cytoplasmic tail, constitutively engages tumor necrosis factor receptor-associated factors. 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. 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. A yeast two-hybrid screen identified TES2 interacting proteins, including the tumor necrosis factor receptor-associated death domain protein (TRADD). TRADD was the only protein that interacted with wild-type TES2 and not with isoleucine-mutated TES2. 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. In transfection assays, TRADD and TES2 synergistically mediated high-level NF-kappaB activation. These results indicate that LMP1 appropriates TRADD to enable efficient long-term lymphoblastoid cell outgrowth. High-level NF-kappaB activation also appears to be a critical component of long-term outgrowth.

latent membrane protein 1

tumor necrosis factor receptor

latent membrane protein 1

tumor necrosis factor receptor-associated death domain protein

TRADD

TRADD

TRADD

TRADD

TRADD

TRADD

engages

latent membrane protein 1

interacting

TRADD

interacted

TRADD

associated

TRADD

associated

TRADD

associated

TRADD

activated

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Transcription factor NF-kappaB regulates inducible Oct-2 gene expression in precursor B lymphocytes.
The POU transcription factors Oct-1 and Oct-2 regulate the activity of octamer-dependent promoters, including those that direct transcription from rearranged immunoglobulin genes. Unlike Oct-1, which is constitutively expressed in many cell types, Oct-2 expression is restricted primarily to B lymphocytes and can be induced in precursor B cells by stimulation with bacterial lipopolysaccharide (LPS). However, the precise factors that mediate this induction mechanism remain unknown. In the present study, we monitored Oct-2 expression in cells arrested for the activation of NF-kappaB, an LPS-responsive member of the Rel transcription factor family. Despite stimulation with LPS, disruption of the NF-kappaB signaling pathway in precursor B cells led to the loss of inducible Oct-2 DNA binding activity in vitro and the suppression of Oct-2-directed transcription in vivo. This biochemical defect correlated with a specific block to Oct-2 gene expression at the level of transcription, whereas the expression of Oct-1 was unaffected. The finding that Oct-2 is under NF-kappaB control highlights an important cross-talk mechanism involving two distinct transcription factor families that regulate B lymphocyte function.

binding

gene expression

expressed

expression

expression

expression

Despite

loss

loss

binding

block

expression at the level of transcription

unaffected

expression

inducible

gene expression

restricted

expressed

restricted

expression

induced

expression

mediate

induced

inducible

binding

regulates

gene expression

control

expression at the level of transcription

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Paternal expression of WT1 in human fibroblasts and lymphocytes.
The Wilms' tumor suppressor gene ( WT1 ) was previously identified as being imprinted, with frequent maternal expression in human placentae and fetal brains. We examined the allele-specific expression of WT1 in cultured human fibroblasts from 15 individuals. Seven of 15 fibroblast lines were heterozygous for polymorphic alleles, and the expression patterns were variable, i.e., equal, unequal or monoallelic paternal expression in three, two and two cases, respectively. Exclusive paternal expression of WT1 was also shown in non-cultured peripheral lymphocytes from the latter two individuals. The allele-specific expression profiles of other imprinted genes, IGF2 and H19, on human chromosome 11 were constant and consistent with those in other tissues. Our unexpected observations of paternal or biallelic expression of WT1 in fibroblasts and lymphocytes, together with the previous findings of maternal or biallelic expression in placentae and brains, suggest that the allele-specific regulatory system of WT1 is unique and may be controlled by a putative tissue- and individual-specific modifier.

Paternal expression

maternal expression

allele-specific expression

expression

paternal expression

allele-specific expression

allele-specific expression

biallelic expression

expression

biallelic expression

expression

allele-specific regulatory

controlled

allele-specific regulatory

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Extinction of immunoglobulin gene expression in B cells upon fusion with HeLa cells is preceded by rapid nuclear depletion of essential transcription factors and is accompanied by widespread inactivation of genes expressed in a B cell-specific manner.
When immunoglobulin (Ig) expressing B cells are fused with non-B cells, Ig expression is rapidly suppressed at the level of transcription, a phenomenon termed extinction. Here we demonstrate that fusion of HeLa cells with either diploid or tetraploid B cells (Daudi) results in widespread extinction of several other B cell-encoded genes that are expressed in a B cell-specific manner. In contrast, expression of B cell-expressed genes that are not dependent on cell-specific controls is unaffected. We show that the molecular mechanism(s) underlying Ig gene extinction can be explained, at least in part, by a lack of transcription factors that are essential for Ig gene transcription. These transcription factors are either not produced due to block of transcription of their respective genes (Oct-2, OBF-1, PU.1), or are rendered inactive posttranslationally (NF-kappa B, E47). By isolating Daudi x HeLa heterokaryons a few hours after fusion, we have studied the initial fate of two B cell-specific transcription factors involved in Ig gene transcription, Oct-2 and NF-kappa B. This report provides the first demonstration that upon fusion with HeLa cells, the nuclear contents of B cell-expressed transcription factors are depleted within a few hours with kinetics that are as fast or faster than that of Ig gene extinction. Thus, the extinguishing mechanism is effective very early after fusion. We suggest that extinction of Ig genes is part of a global mechanism that suppresses the differentiation program foreign to the HeLa phenotype.

block

transcription

inactive

transcription

transcription

transcription

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Regulation of Id3 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. 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. 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). 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. 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. 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. 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.

cyclin E

cyclin A

phosphorylation

phosphorylation

phosphorylation

dependent

phosphorylation

dependent

cyclin E

phosphorylation

dependent

phosphorylation

dependent

cyclin A

phosphorylation

dependent

phosphorylation

Regulation

phosphorylation

regulated

phosphorylation

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Cytokine rescue from glucocorticoid induced apoptosis in T cells is mediated through inhibition of IkappaBalpha.
We previously reported that dexamethasone (DEX), a synthetic glucocorticoid, causes apoptosis in mature Th cell lines, and that this induction of cell death is prevented by specific cytokines, namely, by IL-2 in Th1 cells and by IL-4 in Th2 cells. We now show that this differential rescue by specific cytokines in Th cells correlates with the level of IkappaBalpha that is regulated by DEX and cytokines. In both cell types the cellular levels of IkappaBalpha mRNA and protein were evaluated by DEX treatment. Interestingly, the DEX-mediated IkappaBalpha induction was completely inhibited by IL-2, but not IL-4, in Th1 cells, while the reverse profile was seen in Th2 cells. In both cell types, the cytokine that inhibits the induction of IkappaBalpha by DEX, also rescues these cells from DEX-induced apoptosis, although the rescue cytokine is different in Th1 and Th2 cells. Our results imply that T cells need to maintain a certain level of NF-kappaB transcriptional activity in order to survive; up- or down-regulation of nuclear NF kappaB through modulation of IkappaBalpha expression by cytokines or DEX may lead to cell survival or cell death, respectively.

IkappaBalpha

IkappaBalpha

IkappaBalpha

IkappaBalpha

IkappaBalpha

IkappaBalpha

level

IkappaBalpha

levels

IkappaBalpha

expression

IkappaBalpha

inhibition

IkappaBalpha

inhibited

induction

inhibited

induction

inhibits

induction

induction

IkappaBalpha

induction

IkappaBalpha

regulated

level

modulation

expression

levels

IkappaBalpha

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Xenogeneic human serum promotes leukocyte adhesion to porcine endothelium under flow conditions, possibly through the activation of the transcription factor NF-kappa B.
Endothelial cell activation and leukocyte infiltration are a consistent feature of discordant xenograft rejection. Here we evaluated whether xenogeneic serum, as a source of xenoreactive natural antibodies and complement, induced endothelial cell activation with consequent leukocyte adhesion under flow conditions. Porcine aortic endothelial cells (PAEC) were incubated for 1 hr 30 min or 5 hr with 10% homologous porcine serum (control) or 10% xenogeneic human serum and then perfused with total human leukocytes in a parallel plate flow chamber under laminar flow (1.5 dynes/cm2). Adherent cells were counted by digital image analysis. Xenogeneic human serum significantly (P < 0.01) increased the number of adherent leukocytes as compared with porcine serum. A similar adhesive response was elicited by TNF alpha (100 U/ml), one of the most potent inducers of endothelial cell adhesive properties, here used as positive control. In order to elucidate possible mechanisms underlying endothelial cell activation by xenogeneic serum, we focussed on transcription factor NF-kappa B, a central regulator for the induction of different genes, including adhesive molecules and chemoattractants. By confocal fluorescence microscopy, we observed a positive staining for NF-kappa B (p65 subunit) in the nuclei of PAEC exposed for 1 hr 30 min to human serum, which indicated NF-kappa B activation in this setting. At variance, in PAEC incubated with the homologous serum, NF-kappa B was strictly localized in the cell cytoplasm. Treatment of PAEC exposed to xenogeneic serum with the NF-kappa B inhibitors pyrrolidinedithiocarbamate (PDTC, 25 microM) and tosyl-phechloromethylketone (TPCK, 25 microM) significantly (P < 0.01) reduced leukocyte adhesion in respect to PAEC treated with human serum alone. Findings that xenogeneic serum promotes leukocyte-endothelium interaction possibly through NF-kappa B activation might be relevant for designing future therapeutic strategies aimed at prolonging xenograft survival.

positive staining

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
IL-2-induced growth of CD8+ T cell prolymphocytic leukemia cells mediated by NF-kappaB induction and IL-2 receptor alpha expression.
The binding of interleukin-2 (IL-2) to its receptor on normal T cells induces nuclear expression of nuclear factor kappaB (NF-kappaB), activation of the IL-2 receptor (IL-2R) alpha chain gene, and cell proliferation. In the present study, the role of IL-2R signaling in the growth of CD8+ T cell prolymphocytic leukemia (T-PLL) cells has been investigated. Flow cytometry revealed that primary leukemia cells from a patient with CD8+ T-PLL expressed IL-2Ralpha and beta chains, and the cells showed a proliferative response and an increase in IL-2Ralpha expression on culture with exogeneous IL-2. Northern blot analysis failed to detect IL-2 mRNA, suggesting that IL-2 may act in a paracrine manner in vivo. Electrophoretic mobility-shift assays revealed that recombinant IL-2 increased NF-kappaB binding activity in nuclear extracts of the leukemia cells, and Northern blot analysis showed that IL-2 increased the abundance of mRNAs encoding the NF-kappaB components c-Rel and KBF1 in these cells. IL-2 binding analysis demonstrated that IL-2 markedly increased the number of low affinity IL-2Rs on the leukemia cells, without an effect on the number of high-affinity IL-2Rs. These results show that IL-2 is capable of inducing the nuclear expression of NF-kappaB in primary CD8+ T-PLL cells, and that this effect is mediated, at least in part, at a pretranslational level.

IL-2 receptor alpha

IL-2 receptor (IL-2R) alpha chain

beta

c-Rel

binding

binding

expression

IL-2 receptor alpha

expressed

expressed

beta

expression

induces

binding

activation

activation

IL-2 receptor (IL-2R) alpha chain

increase

expression

increased

abundance of mRNAs

detect

abundance of mRNAs

c-Rel

abundance of mRNAs

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Molecular and cellular analysis of human immunodeficiency virus-induced apoptosis in lymphoblastoid T-cell-line-expressing wild-type and mutated CD4 receptors.
We have previously shown that the presence of the CD4 cytoplasmic tail is critical for human immunodeficiency virus (HIV)-induced apoptosis (J.Corbeil, M.Tremblay, and D.D.Richman, J.Exp.Med.183:39-48, 1996). We have pursued our investigation of the role of the CD4 transduction pathway in HIV-induced apoptosis. To do this, wild-type and mutant forms of the CD4 cytoplasmic tail were stably expressed in the lymphoblastoid T-cell line A2.01. Apoptosis was prevented when CD4 truncated at residue 402 was expressed; however, cells expressing mutated receptors that do not associate with p56(lck) (mutated at the dicysteine motif and truncated at residue 418) but which conserved proximal domains of the cytoplasmic tail underwent apoptosis like wild- type CD4. The differences between wild-type and mutated receptors in the induction of apoptosis were not related to levels of p56(lck) or NF- kappaB activation. Initial signaling through the CD4 receptor played a major role in the sensitization of HIV-infected T cells to undergo apoptosis. Incubation of HIV-infected cells with monoclonal antibody (MAb) 13B8-2, which binds to CD4 in a region critical for dimerization of the receptor, prevented apoptosis without inhibiting HIV replication. Moreover, the apoptotic process was not related to Fas-Fas ligand interaction; however, an antagonistic anti-Fas MAb (ZB-4) enhanced apoptosis in HIV-infected cells without inducing apoptosis in uninfected cells. These observations demonstrate that CD4 signaling mediates HIV-induced apoptosis by a mechanism independent of Fas-Fas ligand interaction, does not require p56(lck) signaling, and may involve a critical region for CD4 dimerization.

CD4 receptor

CD4 receptor

Fas

Fas ligand

Fas

Fas

Fas ligand

associate

binds

interaction

Fas

Fas ligand

interaction

Fas

Fas ligand

dimerization

expressing

CD4 receptor

expressed

expressed

expressing

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
NF-kappaB only partially mediates Epstein-Barr virus latent membrane protein 1 activation of B cells.
The latent membrane protein 1 (LMP1) of Epstein-Barr virus (EBV) is required for EBV-induced immortalization of human B cells and causes tumorigenic transformation of cell lines. LMP1 expression induces phenotypic changes resembling B cell activation, such as cell size increase and up-regulation of cell surface activation markers. LMP1 contains two domains that activate the transcription factor NF-kappaB, one through interactions with TRAF proteins and the other with the TRADD protein. The purpose of the present study was to investigate the importance of NF-kappaB induction in the up-regulation of the B cell activation markers ICAM-1 and CD71 by LMP1. This study shows that expression of LMP1 activates transcription from p50/p65- and c-Rel- responsive promoters, and that this activity can be completely inhibited by expression of a dominant inhibitory IkappaB mutant. ICAM-1 and CD71 are nevertheless up-regulated by LMP1 in primary B cells and cell lines expressing the dominant IkappaB. Furthermore, LMP1-induced cell size increase of primary B cells was unaffected by IkappaB expression. It was concluded that even when LMP1 is unable to activate NF-kappaB, it is still capable of inducing certain characteristics of activated B cells, strongly suggesting that LMP1 can also activate cells independently of NF-kappaB.

latent membrane protein 1

latent membrane protein 1

c-Rel

expression

expression

up-regulation

up-regulation

up-regulated

up-regulated

importance

up-regulation

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Relationship between IkappaBalpha constitutive expression, TNFalpha synthesis, and apoptosis in EBV-infected lymphoblastoid cells.
In order to understand the role of NF-kappaB in EBV transformation we have established stably transfected IkappaBalpha into lymphoblastoid cells. Two clones were obtained in which the loss of NF-kappaB binding activity correlated with the constitutive expression of the transgenic IkappaBalpha. Protein latency expression was determined by immunocytochemistry. Expression of surface markers, intracytoplasmic content of cytokines cell cycle analysis after BrdU incorporation and DNA staining with propidium iodide were studied by flow cytometry. Percentage of apoptotic cells was determined by in-situ labelling of DNA strand breaks. No significative changes in EBV latency nor in cell surface marker expression was found. In contrast, intracytoplasmic TNFalpha levels were strongly reduced in transfected clones. Furthermore, 30% of IkappaBalpha transfected cells were apoptotic after 8 h of TNFalpha treatment. This correlated with a strong reduction of BrdU incorporation after 24 h of TNFalpha treatment. No effect was seen with non transfected cells or with cells transfected with a control plasmid. Our results suggest that the TNFalpha gene could be one of the targets of NF-kappaB in EBV infected cells and that NF-kappaB protects EBV-infected cells from apoptosis induced by TNFalpha, which may favour the proliferative effect of this cytokine.

IkappaBalpha

TNFalpha

IkappaBalpha

IkappaBalpha

TNFalpha

IkappaBalpha

TNFalpha

TNFalpha

TNFalpha

TNFalpha

expression

IkappaBalpha

synthesis

TNFalpha

stably transfected

IkappaBalpha

constitutive expression

IkappaBalpha

transfected

IkappaBalpha

reduced

TNFalpha

stably transfected

stably transfected

transfected

transfected

targets

TNFalpha

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Uncoupling activation-dependent HS1 phosphorylation from nuclear factor of activated T cells transcriptional activation in Jurkat T cells: differential signaling through CD3 and the costimulatory receptors CD2 and CD28.
CD3, CD2, and CD28 are functionally distinct receptors on T lymphocytes. Engagement of any of these receptors induces the rapid tyrosine phosphorylation of a shared group of intracellular signaling proteins, including Vav, Cbl, p85 phosphoinositide 3-kinase, and the Src family kinases Lck and Fyn. Ligation of CD3 also induces the tyrosine phosphorylation of HS1, a 75-kDa hematopoietic cell-specific intracellular signaling protein of unknown function. We have examined changes in HS1 phosphorylation after differential stimulation of CD3, CD2, and CD28 to elucidate its role in T cells and to further delineate the signaling pathways recruited by these receptors. Unlike ligation of CD3, stimulation with anti-CD28 mAb or CHO cells expressing the CD28 ligands CD80 or CD86 did not lead to tyrosine phosphorylation of HS1 in Jurkat T cells. Additionally, no tyrosine phosphorylation of HS1 was induced by mitogenic pairs of anti-CD2 mAbs capable of activating the transcription factor NFAT (nuclear factor of activated T cells). Costimulation through CD28 and/or CD2 did not modulate the CD3-dependent phosphorylation of HS1. In vivo studies indicated that CD3-induced HSI phosphorylation was dependent upon both the Src family tyrosine kinase Lck and the tyrosine phosphatase CD45, did not require MEK1 kinase activity, and was regulated by protein kinase C activation. Thus, although CD3, CD28, and CD2 activate many of the same signaling molecules, they differed in their capacity to induce the tyrosine phosphorylation of HSI. Furthermore, activation-dependent tyrosine phosphorylation of HS1 was not required for NFAT transcriptional activation.

tyrosine

tyrosine

tyrosine

tyrosine

tyrosine

tyrosine

Vav

Cbl

Src

Lck

Fyn

HSI

Src

Lck

HSI

Engagement

Engagement

ligands

ligands

expressing

expressing

phosphorylation

phosphorylation

tyrosine

Vav

phosphorylation

tyrosine

Cbl

phosphorylation

tyrosine

Lck

phosphorylation

tyrosine

Fyn

phosphorylation

tyrosine

phosphorylation

phosphorylation

tyrosine

phosphorylation

tyrosine

phosphorylation

phosphorylation

HSI

phosphorylation

tyrosine

HSI

phosphorylation

tyrosine

dependent

phosphorylation

induces

Engagement

phosphorylation

induces

phosphorylation

stimulation

stimulation

lead

phosphorylation

induced

phosphorylation

dependent

phosphorylation

induced

phosphorylation

dependent

induced

Lck

dependent

induced

require

induced

induce

phosphorylation

induce

phosphorylation

dependent

phosphorylation

changes

phosphorylation

stimulation

modulate

dependent

regulated

induced

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
IL-7 reconstitutes multiple aspects of v-Abl-mediated signaling.
The mechanism by which early lymphoid cells are selectively transformed by v-Abl is currently unknown. Previous studies have shown constitutive activation of IL-4 and IL-7 signaling pathways, as measured by activation of Janus protein kinase (JAK)1, JAK3, STAT5, and STAT6, in pre-B cells transformed by v-Abl. To determine whether activation of these cytokine signaling pathways by v-Abl is important in the cellular events induced by the Abelson murine leukemia virus, the effects of IL-4 and IL-7 on pre-B cells transformed with a temperature-sensitive v-Abl mutant were examined. Whereas IL-4 had little or no effect, IL-7 delayed both the apoptosis and cell cycle arrest that occur upon v-Abl kinase inactivation. IL-7 also delayed the decreases in the levels of c-Myc, Bcl-2, and Bcl-xL that occur upon loss of v-Abl kinase activity. IL-7 did not maintain v-Abl-mediated differentiation arrest of the pre-B cells, as activation of NF-kappaB and RAG gene transcription was unaffected by IL-7. These results identify a potential role for IL-7 signaling pathways in transformation by v-Abl while demonstrating that a combination of IL-4 and IL-7 signaling cannot substitute for an active v-Abl kinase in transformed pre-B cells.

v-Abl

v-Abl

Janus protein kinase (JAK)1

v-Abl

v-Abl

v-Abl

c-Myc

Bcl-xL

v-Abl

v-Abl

v-Abl

delayed

decreases

decreases

c-Myc

decreases

decreases

Bcl-xL

reconstitutes

signaling

signaling

v-Abl

activation

Janus protein kinase (JAK)1

activation

activation

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
p21ras initiates Rac-1 but not phosphatidyl inositol 3 kinase/PKB, mediated signaling pathways in T lymphocytes.
p21ras is activated by the T cell antigen receptor (TCR) and then co-ordinates important signaling pathways for T lymphocyte activation. Effector pathways for this guanine nucleotide binding protein in T cells are mediated by the serine/threonine kinase Raf-1 and the Ras-related GTPase Rac-1. In fibroblasts, an important effector for the Ras oncogene is Phosphatidylinositol 3-kinase (PtdIns 3-kinase). Activation of this lipid kinase is able to induce critical Rac-1 signaling pathways and can couple p21ras to cell survival mechanisms via the serine/threonine kinase Akt/PKB. The role of PtdIns 3-kinase in Ras signaling in T cells has not been explored. In the present study, we examined the ability of PtdIns 3-kinase to initiate the Rac-1 signaling pathways important for T cell activation. We also examined the possibility that Akt/PKB is regulated by Ras signaling pathways in T lymphocytes. The results show that Ras can initiate a Rac-1 mediated pathway that regulates the transcriptional function of AP-1 complexes. PtdIns 3-kinase signals cannot mimic p21ras and induce the Rac mediated responses of AP-1 transcriptional activation. Moreover, neither TCR or Ras activation of AP-1 is dependent on PtdIns 3-kinase. PKB is activated in response to triggering of the T cell antigen receptor; PtdIns 3-kinase activity is both required and sufficient for this TCR response. In contrast, p21ras signals are unable to induce Akt/PKB activity in T cell nor is Ras function required for Akt/PKB activation in response to the TCR. The present data thus highlight that PtdIns 3-kinase and Akt/PKB are not universal Ras effector molecules. Ras can initiate Rac-1 regulated signaling pathways in the context of T cell antigen receptor function independently of PtdIns 3-kinase activity.

PKB

Akt

PKB

Akt

PKB

PKB

Akt

PKB

Akt

PKB

Akt

PKB

activated

mediated

Effector pathways

mediated

Effector pathways

couple

Akt

activated

PKB

required and sufficient

activated

induce

Akt

required

activation

activation

Akt

Effector pathways

regulated

Akt

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Signal transduction abnormalities in T lymphocytes from patients with advanced renal carcinoma: clinical relevance and effects of cytokine therapy.
Studies have demonstrated abnormalities of the CD3/T-cell antigen receptor (TCR) and pathways of signal transduction in T lymphocytes from animals and patients with advanced malignancy. Diminished expression of TCRzeta and p56(lck) that are associated with the TCR and reduced nuclear localization of RelA containing nuclear factor kappaB (NFkappaB) complexes have been noted. These defects have been described in T cells from patients with malignant melanoma, renal cell carcinoma (RCC), ovarian cancer, and colorectal cancer. Preliminary observations also indicate possible correlation with clinical variables such as stage in selected instances. To further characterize altered expression of TCRzeta, p56(lck), and impaired activation of NFkappaB, T lymphocytes were obtained from 65 patients with RCC, the majority of whom were receiving combination cytokine therapy [interleukin (IL)-2, IFN alpha-containing regimens] and 37 control individuals. In 29 of these patients, levels of TCRzeta and p56(lck) were determined by Western blots of T-cell lysates and semiquantitated using densitometry. Relative levels were then correlated with a series of clinical variables including response to therapy, performance status, survival, disease sites, age, and others. In another group of 28 patients (three individuals from the first group), the frequency of abnormal NFkappaB activation was studied using electrophoretic mobility shift assays after activation of T cells with phorbol myristate acetate/ionomycin or anti-CD3 monoclonal antibody. Changes in these signaling molecules during cytokine treatment were also investigated. TCRzeta and p56(lck) were detected in the peripheral blood T cells in 27 of 29 patients, and overall, reduced levels were noted visually in 12 of 29 (41%) and 13 of 29 (45%) individuals, respectively. When levels were semiquantitated using densitometry, significant decreases of TCRzeta (P = 0.029) and p56(lck) (P = 0.029) but not CD3epsilon (P = 0.131), compared with control levels, were found. In patients treated with IL-2/IFN alpha-based therapy, relative levels of TCRzeta increased significantly (P = 0.002) on day 15 of cycle one compared with the baseline. Correlations of TCRzeta or p56(lck) levels with response or disease variables, except for lower TCRzeta levels (P < 0.001) in the presence of bone metastases, were not found. Abnormal NFkappaB activation after stimulation with phorbol myristate acetate/ionomycin and/or anti-CD3 monoclonal antibody was found in 59% of patients (17 of 28) and was not accounted for by the advanced age of the study cohort. Activation of NFkappaB in peripheral blood T cells was inducible during cytokine therapy in four of six individuals who displayed impaired NFkappaB activity prior to therapy. Moreover, impaired activation of NFkappaB does not appear linked to a reduction of TCRzeta expression, because in five patients, normal TCRzeta levels were present although kappaB binding was not inducible. In the majority of patients with advanced RCC, peripheral blood T cells express TCRzeta and p56(lck), and in a subset, reduced levels of these TCRzeta associated molecules are seen that may increase during cytokine-based therapy. Abnormal activation of NFkappaB is also present in >50% of patients and may also revert to normal during IL-2/IFN alpha-based treatment. This alteration in NFkappaB activation occurred in the presence of normal expression of TCRzeta-associated signaling elements. The clinical significance of these findings remains unclear.

nuclear

TCRzeta

RelA

TCRzeta

interleukin (IL)-2

IFN alpha

TCRzeta

TCRzeta

TCRzeta

IFN alpha

TCRzeta

TCRzeta

TCRzeta

TCRzeta

TCRzeta

TCRzeta

TCRzeta

FN alpha

TCRzeta

associated

TCRzeta

associated

expression

TCRzeta

expression

expression

TCRzeta

expression

levels

TCRzeta

levels

detected

TCRzeta

detected

expression

TCRzeta

express

TCRzeta

express

localization

nuclear

RelA

Diminished

expression

reduced

localization

defects

Diminished

defects

reduced

altered

expression

reduced

detected

decreases

TCRzeta

decreases

decreases

lower

TCRzeta

reduction

expression

reduced levels

TCRzeta

reduced levels

Changes

TCRzeta

Changes

increased

TCRzeta

increase

reduced levels

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Transcriptional regulation of lysosomal acid lipase in differentiating monocytes is mediated by transcription factors Sp1 and AP-2.
Human lysosomal acid lipase (LAL) is a hydrolase required for the cleavage of cholesteryl esters and triglycerides derived from plasma lipoproteins. It is shown here that during monocyte to macrophage differentiation, the expression of LAL-mRNA is induced. This induction is dependent on protein kinase C activity and protein synthesis. The cell type-specific increase in LAL expression is further investigated in the THP-1 cell line with respect to transcriptional regulation. The human monocytic leukemia cell line THP-1 differentiates into macrophage-like cells when treated with phorbol esters. In order to determine the cis-acting elements necessary for both basal and phorbol 12-myristate-13 acetate (PMA)-enhanced promoter activity, we performed deletion analysis and reporter gene assays. A PMA responsive element has been identified between -182 bp and -107 bp upstream of the major transcription start site. Gel mobility shift assays demonstrated that binding of Sp1 and AP-2 to the LAL promoter is increased by PMA in THP-1 cells. Co-transfections with expression plasmids for Sp1 and AP-2 further emphasized the important role of these transcription factors in both basal and PMA-enhanced LAL expression. Our data suggest that differentiation dependent increase of lysosomal acid lipase (LAL) expression in THP-1 cells is mediated by a concerted action of Sp1 and AP-2.

promoter

lysosomal acid lipase

lysosomal acid lipase

LAL

LAL

LAL

LAL

LAL

lysosomal acid lipase

LAL

binding

LAL

promoter

binding

LAL

promoter

expression

LAL

expression

LAL

expression

LAL

induced

expression

dependent

induced

increase

expression

increased

binding

enhanced

expression

increase

expression

Transcriptional regulation

lysosomal acid lipase

mediated

Transcriptional regulation

transcriptional regulation

LAL

important role

enhanced

important role

expression

dependent

increase

mediated

dependent

expression

LAL

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
A nongenomic mechanism for progesterone-mediated immunosuppression: inhibition of K+ channels, Ca2+ signaling, and gene expression in T lymphocytes.
The mechanism by which progesterone causes localized suppression of the immune response during pregnancy has remained elusive. Using human T lymphocytes and T cell lines, we show that progesterone, at concentrations found in the placenta, rapidly and reversibly blocks voltage-gated and calcium-activated K+ channels (KV and KCa, respectively), resulting in depolarization of the membrane potential. As a result, Ca2+ signaling and nuclear factor of activated T cells (NF-AT)-driven gene expression are inhibited. Progesterone acts distally to the initial steps of T cell receptor (TCR)-mediated signal transduction, since it blocks sustained Ca2+ signals after thapsigargin stimulation, as well as oscillatory Ca2+ signals, but not the Ca2+ transient after TCR stimulation. K+ channel blockade by progesterone is specific; other steroid hormones had little or no effect, although the progesterone antagonist RU 486 also blocked KV and KCa channels. Progesterone effectively blocked a broad spectrum of K+ channels, reducing both Kv1.3 and charybdotoxin-resistant components of KV current and KCa current in T cells, as well as blocking several cloned KV channels expressed in cell lines. Progesterone had little or no effect on a cloned voltage-gated Na+ channel, an inward rectifier K+ channel, or on lymphocyte Ca2+ and Cl- channels. We propose that direct inhibition of K+ channels in T cells by progesterone contributes to progesterone-induced immunosuppression.

reducing

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Role of IKK1 and IKK2 in lipopolysaccharide signaling in human monocytic cells.
Mononuclear phagocytes play a major role in immune and inflammatory responses. Bacterial lipopolysaccharide (LPS) induces monocytes to express a variety of genes by activating the NF-kappaB/Rel transcription factor family. Recently, we have reported that the tumor necrosis factor and interleukin 1 signaling pathways activate two kinases, IKK1 and IKK2. Phosphorylation of the IkappaB cytoplasmic inhibitors, IkappaBalpha, IkappaBbeta, and IkappaBepsilon, by these kinases triggers proteolytic degradation and the release of NF-kappaB/Rel proteins into the nucleus. At present, the role of the IKKs in LPS signaling has not been investigated. Here, we report that LPS induces IKK activity in human monocytes and THP-1 monocytic cells. The kinetics of activation of kinase activity in monocytic cells are relatively slow with maximal activity observed at 60 min, which coincides with the degradation of IkappaBs and the nuclear translocation of NF-kappaB. In transfection experiments, overexpression of wild type IKK1, a dominant negative mutant IKK1 (K44M), or wild type IKK2 did not affect LPS-induced kappaB-dependent transcription in monocytic cells. In contrast, a dominant negative mutant of IKK2 inhibited LPS induction of kappaB-dependent transcription in a dose-dependent manner. These results indicate that LPS induction of kappaB-dependent gene expression in human monocytic cells requires activation of IKK2.

IkappaBalpha

IkappaBbeta

IkappaBepsilon

overexpression

overexpression

Phosphorylation

IkappaBalpha

Phosphorylation

IkappaBbeta

Phosphorylation

IkappaBepsilon

activate

activate

by

Phosphorylation

by

Phosphorylation

triggers

by

proteolytic degradation

overexpression

overexpression

activation

proteolytic degradation

IkappaBalpha

proteolytic degradation

IkappaBbeta

proteolytic degradation

IkappaBepsilon

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.


Input:
Effects of overexpression of IL-1 receptor-associated kinase on NFkappaB activation, IL-2 production and stress-activated protein kinases in the murine T cell line EL4.
The association and activation of the IL-1 receptor-associated protein kinase (IRAK) to the IL-1 receptor complex is one of the earliest events detectable in IL-1 signal transduction. We generated permanent clones of the murine T cell line EL4 6.1 overexpressing human (h)IRAK to evaluate the role of this kinase in IL-1 signaling. Overexpression of hIRAK enhanced IL-1-stimulated activation of the transcription factor NFkappaB, whereas a truncated form (N-IRAK) specifically inhibited IL-1-dependent NFkappaB activity. In clones stably overexpressing hIRAK a weak constitutive activation of NFkappaB correlated with a low basal IL-2 production which was enhanced in an IL-1-dependent manner. Compared to the parental cell line the dose-response curve of IL-1-induced IL-2 production was shifted in both potency and efficacy. These results demonstrate that IRAK directly triggers NFkappaB-mediated gene expression in EL4 cells. Qualitatively different effects were observed for the IL-1-induced activation of stress-activated protein (SAP) kinases: permanent overexpression of IRAK did not affect the dose dependence but prolonged the kinetics of IL-1-induced activation of SAP kinases, suggesting that this signaling branch may be regulated by distinct mechanisms.

production

production

production

enhanced

production

induced

production

Effects

production

A generic non-protein physical entity used as an event site or location argument.

A gene or gene-product mention, represented by the GENIA Protein label.

Association of two or more biological participants.

The site on the theme where the event occurs; Site2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme2 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme3 distinguishes an additional filler of the same role.

The primary participant or event affected by the event; Theme4 distinguishes an additional filler of the same role.

Expression or production of a gene product.

Presence or movement of a biological participant at or to a location.

The location at which the event or its theme is situated.

The destination location to which a participant moves.

Regulation that decreases, suppresses, or prevents its theme.

The site on the cause that participates in a regulatory event.

The participant or event responsible for causing or regulating the event.

The site on the theme where the event occurs.

The primary participant or event affected by the event.

Addition of a phosphate group to a biological molecule.

Regulation that increases, promotes, or enables its theme.

Degradation or catabolic processing of a protein.

Regulation whose positive or negative direction is not specified.

Transcription of a gene or DNA template into RNA.