To investigate whether such adaptive changes in TR expression occur in humans , we determined the mRNA levels of the hTR beta 1 in various thyroid states .

To investigate whether such adaptive changes in <protein>TR</protein> expression occur in humans , we determined the mRNA levels of the <protein>hTR beta 1</protein> in various thyroid states .


Patients with overt hypo -or hyperthyroidism were enrolled in the study .


Total RNA was isolated from peripheral blood mononuclear cells and hTR beta 1 mRNA levels determined by quantitative competitive reverse transcription PCR .

Total RNA was isolated from <cell_type>peripheral blood mononuclear cells</cell_type> and <rna>hTR beta 1 mRNA</rna> levels determined by quantitative competitive reverse transcription PCR .


For comparison , hTR beta 1 mRNA levels were determined in lymphocytes and normal thyroid tissue of euthyroid patients .

For comparison , <rna>hTR beta 1 mRNA</rna> levels were determined in <cell_type>lymphocytes</cell_type> and normal thyroid tissue of euthyroid patients .


Human TR beta 1 mRNA levels in lymphocytes were 1.8 +/- 0.4 , 1.9 +/- 0.5 , 1.1 +/- 0.4 10 ( -18 ) mol/microgram RNA in hypo- , eu -and hyperthyroid patients , respectively , corresponding to an estimated 0.5 -2 molecules per cell .

<rna>Human TR beta 1 mRNA</rna> levels in <cell_type>lymphocytes</cell_type> were 1.8 +/- 0.4 , 1.9 +/- 0.5 , 1.1 +/- 0.4 10 ( -18 ) mol/microgram RNA in hypo- , eu -and hyperthyroid patients , respectively , corresponding to an estimated 0.5 -2 molecules per cell .


Although the mean hTR beta 1 mRNA levels were 40 % lower in hyperthyroid than in euthyroid subjects , this difference did not reach statistical significance .

Although the mean <rna>hTR beta 1 mRNA</rna> levels were 40 % lower in hyperthyroid than in euthyroid subjects , this difference did not reach statistical significance .


Similar levels of hTR beta 1 mRNA levels were detected in thyroid gland from euthyroid patients .

Similar levels of <rna>hTR beta 1 mRNA</rna> levels were detected in thyroid gland from euthyroid patients .


In summary , we developed an assay for the quantitative determination of hTR beta 1 mRNA levels in small human tissue samples , containing as little as 50 ng of total RNA .

In summary , we developed an assay for the quantitative determination of <rna>hTR beta 1 mRNA</rna> levels in small human tissue samples , containing as little as 50 ng of total RNA .


Absolute hTR beta 1 mRNA levels are very low with an estimated one molecule of mRNA being present in a mononuclear blood cell or thyrocyte .

Absolute <rna>hTR beta 1 mRNA</rna> levels are very low with an estimated one molecule of <rna>mRNA</rna> being present in a <cell_type>mononuclear blood cell</cell_type> or <cell_type>thyrocyte</cell_type> .


No up-regulation of hTR beta 1 was seen in hypothyroid relative to euthyroid patients .

No up-regulation of <protein>hTR beta 1</protein> was seen in hypothyroid relative to euthyroid patients .


However , there is a non-significant trend towards a down-regulation of hTR beta 1 mRNA levels in hyperthyroid patients .

However , there is a non-significant trend towards a down-regulation of <rna>hTR beta 1 mRNA</rna> levels in hyperthyroid patients .


Multiple proteins interact with the nuclear inhibitory protein repressor element in the human interleukin-3 promoter .

Multiple proteins interact with the <dna>nuclear inhibitory protein repressor element</dna> in the <dna>human interleukin-3 promoter</dna> .


T cell expression of interleukin 3 ( IL-3 ) is directed by positive and negative cis-acting DNA elements clustered within 300 base pairs of the transcriptional start site .

T cell expression of <protein>interleukin 3</protein> ( <protein>IL-3</protein> ) is directed by <dna>positive and negative cis-acting DNA elements</dna> clustered within 300 base pairs of the <dna>transcriptional start site</dna> .


A strong repressor element , termed nuclear inhibitory protein ( NIP ) , was previously mapped to a segment of the IL-3 promoter between nucleotides -271 and -250 .

A strong <protein>repressor element</protein> , termed <protein>nuclear inhibitory protein</protein> ( <protein>NIP</protein> ) , was previously mapped to a segment of the <dna>IL-3 promoter</dna> between <dna>nucleotides -271 and -250</dna> .


Functional characterization of this element demonstrates that it can mediate repression when linked in cis to a heterologous promoter .

Functional characterization of this element demonstrates that it can mediate repression when linked in cis to a <dna>heterologous promoter</dna> .


DNA binding experiments were carried out to characterize the repressor activity .


Using varying conditions , three distinct complexes were shown to interact specifically with the NIP region , although only one correlates with repressor activity .

Using varying conditions , three distinct complexes were shown to interact specifically with the <dna>NIP region</dna> , although only one correlates with repressor activity .


Complex 1 results from binding of a ubiquitous polypeptide that recognizes the 3 ' portion of this sequence and is not required for repression .

<protein>Complex 1</protein> results from binding of a ubiquitous polypeptide that recognizes the <dna>3 ' portion</dna> of this sequence and is not required for repression .


Complex 2 corresponds to binding of transcription factor ( upstream stimulatory factor ) to an E-box motif in the 5 ' portion of the NIP region .

<protein>Complex 2</protein> corresponds to binding of <protein>transcription factor</protein> ( <protein>upstream stimulatory factor</protein> ) to an <dna>E-box motif</dna> in the <dna>5 ' portion</dna> of the <dna>NIP region</dna> .


DNA binding specificity of complex 3 overlaps with that of upstream stimulatory factor but is clearly distinct .

DNA binding specificity of complex 3 overlaps with that of <protein>upstream stimulatory factor</protein> but is clearly distinct .


To determine which of the latter two complexes represents NIP activity , we incorporated small alterations into the NIP site of an IL-3 promoter-linked reporter construct and examined their effects on NIP -mediated repression .

To determine which of the latter two complexes represents <protein>NIP</protein> activity , we incorporated small alterations into the <dna>NIP site</dna> of an <dna>IL-3 promoter-linked reporter construct</dna> and examined their effects on <protein>NIP</protein> -mediated repression .


Functional specificity for repression matches the DNA binding specificity of complex 3 ; both repressor activity and complex 3 binding require the consensus sequence CTCACNTNC .

Functional specificity for repression matches the DNA binding specificity of <protein>complex 3</protein> ; both repressor activity and <protein>complex 3</protein> binding require the <dna>consensus sequence</dna> CTCACNTNC .


The hematopoietic transcription factor PU.1 is downregulated in human multiple myeloma cell lines .

The <protein>hematopoietic transcription factor</protein> <protein>PU.1</protein> is downregulated in <cell_line>human multiple myeloma cell lines</cell_line> .


PU.1 is a hematopoietic transcription factor belonging to the Ets-family .

<protein>PU.1</protein> is a <protein>hematopoietic transcription factor</protein> belonging to the <protein>Ets-family</protein> .


It is identical to the Spi-1 oncogene , which is implicated in spleen focus-forming virus-induced murine erythroleukemias .

It is identical to the <dna>Spi-1 oncogene</dna> , which is implicated in spleen focus-forming virus-induced murine erythroleukemias .


PU.1 seems to be required for early development of multiple hematopoietic lineages , but its expression in mature cells is preferentially observed in cells of the B-cell-and monocyte/macrophage-differentiation lineage .

<protein>PU.1</protein> seems to be required for early development of <cell_type>multiple hematopoietic lineages</cell_type> , but its expression in <cell_type>mature cells</cell_type> is preferentially observed in cells of the <cell_type>B-cell-and monocyte/macrophage-differentiation lineage</cell_type> .


It binds the so-called Pu box , an important tissue-specific regulatory DNA element present in a number of genes expressed in these cell lineages .

It binds the so-called <dna>Pu box</dna> , an important <dna>tissue-specific regulatory DNA element</dna> present in a number of genes expressed in these <cell_type>cell lineages</cell_type> .


We have analyzed the expression and activity of PU.1 during human B-cell development using a panel of B-cell lines representing different stages of maturation , from early precursors to differentiated plasma cells .

We have analyzed the expression and activity of <protein>PU.1</protein> during human B-cell development using a panel of <cell_line>B-cell lines</cell_line> representing different stages of maturation , from <cell_line>early precursors</cell_line> to <cell_line>differentiated plasma cells</cell_line> .


PU.1 mRNA expression and PU.1 DNA binding activity , as measured by Northern blot analysis and electrophoretic mobility shift assay , respectively , were evident in cell lines representing pro-B , pre-B , and mature B cells .

<rna>PU.1 mRNA</rna> expression and <protein>PU.1</protein> DNA binding activity , as measured by Northern blot analysis and electrophoretic mobility shift assay , respectively , were evident in <cell_line>cell lines</cell_line> representing <cell_line>pro-B , pre-B , and mature B cells</cell_line> .


We could also show Pu box -dependent transactivation of a reporter gene in transient transfections in these cell lines .

We could also show <dna>Pu box</dna> -dependent transactivation of a reporter gene in transient transfections in these <cell_line>cell lines</cell_line> .


In contrast , in a number of multiple myeloma cell lines , representing differentiated , plasma cell-like B cells , PU.1 DNA binding activity , mRNA expression , and Pu box -dependent transactivation were absent or detectable at a very low level .

In contrast , in a number of <cell_line>multiple myeloma cell lines</cell_line> , representing <cell_type>differentiated , plasma cell-like B cells</cell_type> , <protein>PU.1</protein> DNA binding activity , mRNA expression , and <dna>Pu box</dna> -dependent transactivation were absent or detectable at a very low level .


In lymphoblastoid cell lines , which exemplify an intermediate stage of B-cell differentiation , a reduced expression and activity were observed .

In <cell_line>lymphoblastoid cell lines</cell_line> , which exemplify an intermediate stage of B-cell differentiation , a reduced expression and activity were observed .


The findings in the human multiple myeloma cell lines represent the first examples of B cells with downregulated PU.1 expression and apparently contradict observations in the murine system in which PU.1 is expressed and active in plasmacytoma cell lines .

The findings in the <cell_line>human multiple myeloma cell lines</cell_line> represent the first examples of <cell_type>B cells</cell_type> with downregulated <protein>PU.1</protein> expression and apparently contradict observations in the murine system in which <protein>PU.1</protein> is expressed and active in <cell_line>plasmacytoma cell lines</cell_line> .


At present , it is unclear whether the lack of PU.1 expression and activity in human multiple myeloma cell lines represents a malignancy-associated defect in these cells or exemplifies a normal developmental regulation in terminally differentiated B cells .

At present , it is unclear whether the lack of <protein>PU.1</protein> expression and activity in <cell_line>human multiple myeloma cell lines</cell_line> represents a malignancy-associated defect in these cells or exemplifies a normal developmental regulation in <cell_type>terminally differentiated B cells</cell_type> .


Regulation of granulocyte-macrophage colony-stimulating factor and E-selectin expression in endothelial cells by cyclosporin A and the T-cell transcription factor NFAT .

Regulation of <protein>granulocyte-macrophage colony-stimulating factor</protein> and <protein>E-selectin</protein> expression in <cell_type>endothelial cells</cell_type> by cyclosporin A and the <protein>T-cell transcription factor</protein> <protein>NFAT</protein> .


Nuclear factor of activated T cells ( NFAT ) was originally described as a T-cell-specific transcription factor athat supported the activation of cytokine gene expression and mediated the immunoregulatory effects of cyclosporin A ( CsA ) .

<protein>Nuclear factor of activated T cells</protein> ( <protein>NFAT</protein> ) was originally described as a <protein>T-cell-specific transcription factor</protein> athat supported the activation of <protein>cytokine</protein> gene expression and mediated the <protein>immunoregulatory</protein> effects of cyclosporin A ( CsA ) .


As we observed that activated endothelial cells also expressed NFAT , we tested the antiinflammatory properties of CsA in endothelial cells .

As we observed that <cell_type>activated endothelial cells</cell_type> also expressed <protein>NFAT</protein> , we tested the antiinflammatory properties of CsA in <cell_type>endothelial cells</cell_type> .


Significantly , CsA completely suppressed the induction of NFAT in endothelial cells and inhibited the activity of granulocyte-macrophage colony-stimulating factor ( GM-CSF ) gene regulatory elements that use NFAT by 60 % .

Significantly , CsA completely suppressed the induction of <protein>NFAT</protein> in <cell_type>endothelial cells</cell_type> and inhibited the activity of <dna>granulocyte-macrophage colony-stimulating factor ( GM-CSF ) gene regulatory elements</dna> that use <protein>NFAT</protein> by 60 % .


CsA similarly mediated a reduction of up to 65 % in GM-CSF mRNA and protein expression in activated endothelial cells .

CsA similarly mediated a reduction of up to 65 % in <rna>GM-CSF mRNA</rna> and protein expression in <cell_type>activated endothelial cells</cell_type> .


CsA also suppressed E-selectin , but not vascular cell adhesion molecule-1 ( VCAM-1 ) expression in endothelial cells , even though the E-selectin promoter is activated by NF-kappa B rather than NFAT .

CsA also suppressed <protein>E-selectin</protein> , but not <protein>vascular cell adhesion molecule-1</protein> ( <protein>VCAM-1</protein> ) expression in endothelial cells , even though the <dna>E-selectin promoter</dna> is activated by <protein>NF-kappa B</protein> rather than <protein>NFAT</protein> .


Hence , induction of cell surface expression of this leukocyte adhesion molecule by tumor necrosis factor ( TNF ) -alpha was reduced by 40 % in the presence of CsA , and this was reflected by a 29 % decrease in neutrophil adhesion .

Hence , induction of cell surface expression of this <protein>leukocyte adhesion molecule</protein> by <protein>tumor necrosis factor ( TNF ) -alpha</protein> was reduced by 40 % in the presence of CsA , and this was reflected by a 29 % decrease in <cell_type>neutrophil</cell_type> adhesion .


The effects of CsA on endothelial cells were also detected at the chromatin structure level , as DNasel hypersensitive sites within both the GM-CSF enhancer and the E-selectin promoter were suppressed by CsA .

The effects of CsA on <cell_type>endothelial cells</cell_type> were also detected at the <dna>chromatin</dna> structure level , as <dna>DNasel hypersensitive sites</dna> within both the <protein>GM-CSF</protein> enhancer and the <dna>E-selectin promoter</dna> were suppressed by CsA .


This represents the first report of NFAT in endothelial cells and suggests mechanisms by which CsA could function as an antiinflammatory agent .

This represents the first report of <protein>NFAT</protein> in <cell_type>endothelial cells</cell_type> and suggests mechanisms by which CsA could function as an antiinflammatory agent .


Costimulation requirement for AP-1 and NF-kappa B transcription factor activation in T cells .

Costimulation requirement for <protein>AP-1</protein> and <protein>NF-kappa B</protein> transcription factor activation in <cell_type>T cells</cell_type> .


The transcriptional activity of the IL-2 promoter requires T-cell costimulation delivered by the TCR and the auxiliary receptor CD28 .

The transcriptional activity of the <dna>IL-2 promoter</dna> requires <cell_type>T-cell</cell_type> costimulation delivered by the <protein>TCR</protein> and the <protein>auxiliary receptor CD28</protein> .


Several transcription factors participate in IL-2 promoter activation , among which are AP-1-like factors and NF-kappa B .

Several <protein>transcription factors</protein> participate in <protein>IL-2</protein> promoter activation , among which are <protein>AP-1-like factors</protein> and <protein>NF-kappa B</protein> .


Protein phosphorylation has an important role in the regulation of these two factors : ( 1 ) it induces the transactivating capacity of the AP-1 protein c-Jun ; and ( 2 ) it is involved in the release of the cytoplasmic inhibitor , I kappa B , from NF-kappa B , allowing translocation of the latter into the nucleus .

Protein phosphorylation has an important role in the regulation of these two factors : ( 1 ) it induces the transactivating capacity of the <protein>AP-1 protein</protein> <protein>c-Jun</protein> ; and ( 2 ) it is involved in the release of the <protein>cytoplasmic inhibitor</protein> , <protein>I kappa B</protein> , from <protein>NF-kappa B</protein> , allowing translocation of the latter into the nucleus .


We have recently shown that both phosphorylation processes require T-cell costimulation .


Furthermore , in activated T cells , the kinetics of the two phosphorylation events are essentially similar .

Furthermore , in <cell_type>activated T cells</cell_type> , the kinetics of the two phosphorylation events are essentially similar .


According to our results , however , the kinases responsible for the two processes are distinct entities .


Whereas TPCK inhibits phosphorylation of I kappa B and , consequently , activation of NF-kappa B , it markedly enhances the activity of JNK , the MAP kinase-related kinase that phosphorylates the transactivation domain of c-Jun .

Whereas <protein>TPCK</protein> inhibits phosphorylation of <protein>I kappa B</protein> and , consequently , activation of <protein>NF-kappa B</protein> , it markedly enhances the activity of <protein>JNK</protein> , the <protein>MAP kinase-related kinase</protein> that phosphorylates the <protein>transactivation domain</protein> of <protein>c-Jun</protein> .


We , therefore , propose the activation scheme presented in FIGURE 3 for T-cell costimulation .


Costimulation results in the activation of a signaling pathway that leads to the simultaneous induction of the two transcription factors , AP-1 and NF-kappa B .

Costimulation results in the activation of a signaling pathway that leads to the simultaneous induction of the two <protein>transcription factors</protein> , <protein>AP-1</protein> and <protein>NF-kappa B</protein> .


Integration of the signals generated by TCR and CD28 engagement occurs along this pathway , which then bifurcates to induce I kappa B phosphorylation and NF-kappa B activation on the one hand , and JNK activation and c-Jun phosphorylation on the other .

Integration of the signals generated by <protein>TCR</protein> and <protein>CD28</protein> engagement occurs along this pathway , which then bifurcates to induce <protein>I kappa B</protein> phosphorylation and <protein>NF-kappa B</protein> activation on the one hand , and <protein>JNK</protein> activation and <protein>c-Jun</protein> phosphorylation on the other .


We are currently engaged in defining where the two signals integrate along the AP-1 / NF-kappa B pathway .

We are currently engaged in defining where the two signals integrate along the <protein>AP-1</protein> / <protein>NF-kappa B</protein> pathway .


Regulation of human immunodeficiency virus type 1 and cytokine gene expression in myeloid cells by NF-kappa B/Rel transcription factors .

Regulation of human immunodeficiency virus type 1 and cytokine gene expression in <cell_type>myeloid cells</cell_type> by <protein>NF-kappa B/Rel</protein> <protein>transcription factors</protein> .


CD4+ macrophages in tissues such as lung , skin , and lymph nodes , promyelocytic cells in bone marrow , and peripheral blood monocytes serve as important targets and reservoirs for human immunodeficiency virus type 1 ( HIV-1 ) replication .

<cell_line>CD4+ macrophages</cell_line> in tissues such as lung , skin , and lymph nodes , <cell_type>promyelocytic cells</cell_type> in bone marrow , and <cell_type>peripheral blood monocytes</cell_type> serve as important targets and reservoirs for human immunodeficiency virus type 1 ( HIV-1 ) replication .


HIV-1-infected myeloid cells are often diminished in their ability to participate in chemotaxis , phagocytosis , and intracellular killing .

HIV-1-infected <cell_type>myeloid cells</cell_type> are often diminished in their ability to participate in chemotaxis , phagocytosis , and intracellular killing .


HIV-1 infection of myeloid cells can lead to the expression of surface receptors associated with cellular activation and/or differentiation that increase the responsiveness of these cells to cytokines secreted by neighboring cells as well as to bacteria or other pathogens .

HIV-1 infection of <cell_type>myeloid cells</cell_type> can lead to the expression of <protein>surface receptors</protein> associated with cellular activation and/or differentiation that increase the responsiveness of these cells to <protein>cytokines</protein> secreted by neighboring cells as well as to bacteria or other pathogens .


Enhancement of HIV-1 replication is related in part to increased DNA-binding activity of cellular transcription factors such as NF-kappa B .

Enhancement of HIV-1 replication is related in part to increased DNA-binding activity of <protein>cellular transcription factors</protein> such as <protein>NF-kappa B</protein> .


NF-kappa B binds to the HIV-1 enhancer region of the long terminal repeat and contributes to the inducibility of HIV-1 gene expression in response to multiple activating agents .

<protein>NF-kappa B</protein> binds to the <dna>HIV-1 enhancer</dna> region of the <dna>long terminal repeat</dna> and contributes to the inducibility of HIV-1 gene expression in response to multiple activating agents .


Phosphorylation and degradation of the cytoplasmic inhibitor I kappa B alpha are crucial regulatory events in the activation of NF-kappa B DNA-binding activity .

Phosphorylation and degradation of the <protein>cytoplasmic inhibitor</protein> <protein>I kappa B</protein> alpha are crucial regulatory events in the activation of <protein>NF-kappa B</protein> DNA-binding activity .


Both N- and C-terminal residues of I kappa B alpha are required for inducer-mediated degradation .

Both N- and C-terminal residues of <protein>I kappa B</protein> alpha are required for inducer-mediated degradation .


Chronic HIV-1 infection of myeloid cells leads to constitutive NF-kappa B DNA-binding activity and provides an intranuclear environment capable of perpetuating HIV-1 replication .

Chronic HIV-1 infection of <cell_type>myeloid cells</cell_type> leads to constitutive <protein>NF-kappa B</protein> DNA-binding activity and provides an intranuclear environment capable of perpetuating HIV-1 replication .


Increased intracellular stores of latent NF-kappa B may also result in rapid inducibility of NF-kappa B -dependent cytokine gene expression .

Increased intracellular stores of <protein>latent NF-kappa B</protein> may also result in rapid inducibility of <protein>NF-kappa B</protein> -dependent <protein>cytokine</protein> gene expression .


In response to secondary pathogenic infections or antigenic challenge , cytokine gene expression is rapidly induced , enhanced , and sustained over prolonged periods in HIV-1-infected myeloid cells compared with uninfected cells .

In response to secondary pathogenic infections or antigenic challenge , <protein>cytokine</protein> gene expression is rapidly induced , enhanced , and sustained over prolonged periods in <cell_line>HIV-1-infected myeloid cells</cell_line> compared with <cell_type>uninfected cells</cell_type> .


Elevated levels of several inflammatory cytokines have been detected in the sera of HIV-1-infected individuals .

Elevated levels of several <protein>inflammatory cytokines</protein> have been detected in the sera of HIV-1-infected individuals .


Secretion of myeloid cell-derived cytokines may both increase virus production and contribute to AIDS-associated disorders .

Secretion of <protein>myeloid cell-derived cytokines</protein> may both increase virus production and contribute to AIDS-associated disorders .


Steroid mediated lysis of lymphoblasts requires the DNA binding region of the steroid hormone receptor .

Steroid mediated lysis of <cell_type>lymphoblasts</cell_type> requires the DNA binding region of the steroid hormone receptor .


Glucocorticoids kill certain types of lymphoblasts , but the mechanisms are unknown .

Glucocorticoids kill certain types of <cell_type>lymphoblasts</cell_type> , but the mechanisms are unknown .


It is clear that sufficient numbers of functional glucocorticoid receptors are required to mediate lysis , but whether they do so through the classical model of steroid hormone activation and modulation of gene expression has not been established .

It is clear that sufficient numbers of functional <protein>glucocorticoid receptors</protein> are required to mediate lysis , but whether they do so through the classical model of steroid hormone activation and modulation of gene expression has not been established .


In this report we have asked which region ( s ) of the steroid receptor are important for mediating lysis in leukemic T lymphoblasts .

In this report we have asked which region ( s ) of the <protein>steroid receptor</protein> are important for mediating lysis in <cell_type>leukemic T lymphoblasts</cell_type> .


CEM-ICR 27 leukemic lymphoblasts , a clone of CEM cells which lack functional glucocorticoid receptors and therefore are neither lysed by dexamethasone nor capable of showing glutamine synthetase induction , were provided with steroid receptors by DNA transfections of various receptor gene constructs .

<cell_line>CEM-ICR 27 leukemic lymphoblasts</cell_line> , a clone of <cell_line>CEM cells</cell_line> which lack functional <protein>glucocorticoid receptors</protein> and therefore are neither lysed by dexamethasone nor capable of showing <protein>glutamine synthetase</protein> induction , were provided with <protein>steroid receptors</protein> by DNA transfections of various receptor gene constructs .


We measured steroid mediated lysis , receptor number and induction of glutamine synthetase in the transfected cells .

We measured steroid mediated lysis , receptor number and induction of <protein>glutamine synthetase</protein> in the <cell_line>transfected cells</cell_line> .


Our results provide evidence that the lysis mechanism in the ICR27 lymphoblasts is restored when functional receptor number is restored .

Our results provide evidence that the lysis mechanism in the <cell_line>ICR27 lymphoblasts</cell_line> is restored when functional receptor number is restored .


The DNA binding region specifying high affinity for GRE sites is required .

The <protein>DNA binding region</protein> specifying high affinity for <dna>GRE sites</dna> is required .


Lysis is mediated by any steroid that allows for activation of the receptor containing such a region .


Our data support the view that steroid-mediated cell death occurs by a process requiring direct interaction of steroid-receptor complexes with the genome .

Our data support the view that steroid-mediated cell death occurs by a process requiring direct interaction of <protein>steroid-receptor complexes</protein> with the genome .


Functional characterization of the murine homolog of the B cell-specific coactivator BOB.1/OBF.1 .

Functional characterization of the <protein>murine homolog</protein> of the <protein>B cell-specific coactivator</protein> <protein>BOB.1/OBF.1</protein> .


B cell-specific transcriptional promoter activity mediated by the octamer motif requires the Oct1 or Oct2 protein and additional B cell-restricted cofactors .

B cell-specific transcriptional promoter activity mediated by the <dna>octamer motif</dna> requires the <protein>Oct1</protein> or <protein>Oct2 protein</protein> and additional <protein>B cell-restricted cofactors</protein> .


One such cofactor , BOB.1/OBF.1 , was recently isolated from human B cells .

One such <protein>cofactor</protein> , <protein>BOB.1/OBF.1</protein> , was recently isolated from <cell_type>human B cells</cell_type> .


Here , we describe the isolation and detailed characterization of the murine homolog .

Here , we describe the isolation and detailed characterization of the <protein>murine homolog</protein> .


Full-length cDNAs and genomic clones were isolated , and the gene structure was determined. Comparison of the deduced amino acids shows 88 % sequence identity between mouse and human BOB.1/OBF.1 .

Full-length <dna>cDNAs</dna> and <dna>genomic clones</dna> were isolated , and the gene structure was determined. Comparison of the deduced amino acids shows 88 % sequence identity between <protein>mouse and human BOB.1/OBF.1</protein> .


The NH2-terminal 126 amino acids of BOB.1/OBF.1 are both essential and sufficient for interaction with the POU domains of either Oct1 or Oct2 .

The <protein>NH2-terminal 126 amino acids</protein> of <protein>BOB.1/OBF.1</protein> are both essential and sufficient for interaction with the <protein>POU domains</protein> of either <protein>Oct1</protein> or <protein>Oct2</protein> .


This protein-protein interaction does not require the simultaneous binding of Oct proteins to DNA , and high resolution footprinting of the Oct -DNA interaction reveals that binding of BOB.1/OBF.1 to Oct1 or Oct2 does not alter the interaction with DNA .

This protein-protein interaction does not require the simultaneous binding of <protein>Oct proteins</protein> to DNA , and high resolution footprinting of the <dna>Oct</dna> -DNA interaction reveals that binding of <protein>BOB.1/OBF.1</protein> to <protein>Oct1</protein> or <protein>Oct2</protein> does not alter the interaction with DNA .


BOB.1/OBF.1 can efficiently activate octamer-dependent promoters in fibroblasts ; however , it fails to stimulate octamer-dependent enhancer activity .

<protein>BOB.1/OBF.1</protein> can efficiently activate octamer-dependent promoters in <cell_type>fibroblasts</cell_type> ; however , it fails to stimulate <dna>octamer-dependent enhancer</dna> activity .


Fusion of subdomains of BOB.1/OBF.1 with the GAL4 DNA binding domain reveals that both NH2- and COOH-terminal domains of BOB.1/OBF.1 contribute to full transactivation function , the COOH-terminal domain is more efficient in this transactivation assay .

Fusion of subdomains of <protein>BOB.1/OBF.1</protein> with the <protein>GAL4 DNA binding domain</protein> reveals that both <protein>NH2- and COOH-terminal domains</protein> of <protein>BOB.1/OBF.1</protein> contribute to full transactivation function , the <protein>COOH-terminal domain</protein> is more efficient in this transactivation assay .


Consistent with the failure of full-length BOB.1/OBF.1 to stimulate octamer-dependent enhancer elements in non B cells , the GAL4 fusions likewise only stimulate from a promoter-proximal position .

Consistent with the failure of <protein>full-length BOB.1/OBF.1</protein> to stimulate <dna>octamer-dependent enhancer elements</dna> in <cell_type>non B cells</cell_type> , the <protein>GAL4 fusions</protein> likewise only stimulate from a promoter-proximal position .


Anti-immunoglobulin M activates nuclear calcium/calmodulin-dependent protein kinase II in human B lymphocytes .

<protein>Anti-immunoglobulin M</protein> activates <protein>nuclear calcium/calmodulin-dependent protein kinase II</protein> in <cell_type>human B lymphocytes</cell_type> .


We and others have previously shown that the nuclear protein , Ets-1 , is phosphorylated in a calcium-dependent manner after ligation of immunoglobulin ( Ig ) M on B lymphocytes .

We and others have previously shown that the <protein>nuclear protein</protein> , <protein>Ets-1</protein> , is phosphorylated in a calcium-dependent manner after ligation of immunoglobulin ( Ig ) M on <cell_type>B lymphocytes</cell_type> .


As this phosphorylation was independent of protein kinase C activity , we tested whether a calcium/calmodulin-dependent protein kinase ( CaM kinase ) might phosphorylate the Ets-1 protein after elevation of intracellular free calcium concentrations .

As this phosphorylation was independent of <protein>protein kinase C</protein> activity , we tested whether a <protein>calcium/calmodulin-dependent protein kinase</protein> ( <protein>CaM kinase</protein> ) might phosphorylate the <protein>Ets-1 protein</protein> after elevation of intracellular free calcium concentrations .


The dephosphorylated form of Ets-1 has been shown to bind to chromatin , suggesting that the operative kinase should be detectable in the nucleus .

The dephosphorylated form of <protein>Ets-1</protein> has been shown to bind to chromatin , suggesting that the <protein>operative kinase</protein> should be detectable in the nucleus .


We prepared nuclear extracts from two human B cell lines in which increased intracellular free calcium levels correlated with increased phosphorylation of the Ets-1 protein .

We prepared nuclear extracts from two <cell_line>human B cell lines</cell_line> in which increased intracellular free calcium levels correlated with increased phosphorylation of the <protein>Ets-1</protein> protein .


Activity of the CaM kinases was determined using a synthetic peptide substrate both in the absence and presence of an inhibitor specific for the CaM kinase family , KN-62 .

Activity of the <protein>CaM kinases</protein> was determined using a synthetic peptide substrate both in the absence and presence of an inhibitor specific for the <protein>CaM kinase family</protein> , KN-62 .


Stimulation of cells with anti-IgM led to increased activity of a nuclear kinase that could phosphorylate the peptide , and this activity was reduced by 10 microM KN-62 .

Stimulation of cells with <protein>anti-IgM</protein> led to increased activity of a <protein>nuclear kinase</protein> that could phosphorylate the peptide , and this activity was reduced by 10 microM KN-62 .


Kinase activity was reduced in lysates preadsorbed using an antibody specific for CaM kinase II .

Kinase activity was reduced in lysates preadsorbed using an <protein>antibody</protein> specific for <protein>CaM kinase II</protein> .


Two-dimensional phosphopeptide maps of the Ets-1 protein from cells incubated with ionomycin or anti-IgM contained two unique phosphopeptides that were absent in untreated cells .

Two-dimensional phosphopeptide maps of the <protein>Ets-1</protein> protein from cells incubated with ionomycin or <protein>anti-IgM</protein> contained two unique phosphopeptides that were absent in untreated cells .


Incubation of isolated Ets-1 protein with purified CaM kinase II produced phosphorylation of peptides that migrated identically to those found in cells incubated with either anti-IgM or ionomycin .

Incubation of <protein>isolated Ets-1 protein</protein> with purified <protein>CaM kinase II</protein> produced phosphorylation of peptides that migrated identically to those found in cells incubated with either <protein>anti-IgM</protein> or ionomycin .


These data suggest a model of signal transduction by the antigen receptor on B lymphocytes in which increased intracellular free calcium can rapidly activate nuclear CaM kinase II , potentially resulting in phosphorylation and regulation of DNA-binding proteins .

These data suggest a model of signal transduction by the <protein>antigen receptor</protein> on <cell_type>B lymphocytes</cell_type> in which increased intracellular free calcium can rapidly activate <protein>nuclear CaM kinase II</protein> , potentially resulting in phosphorylation and regulation of <protein>DNA-binding proteins</protein> .


Transcriptional activation and repression , two properties of the lymphoid-specific transcription factor Oct-2a .

Transcriptional activation and repression , two properties of the <protein>lymphoid-specific transcription factor</protein> <protein>Oct-2a</protein> .


The lymphoid-specific transcription factor Oct-2a contains two transcriptional activation domains which are located within the N-terminal and C-terminal regions .

The <protein>lymphoid-specific transcription factor</protein> <protein>Oct-2a</protein> contains two <protein>transcriptional activation domains</protein> which are located within the <protein>N-terminal</protein> and <protein>C-terminal regions</protein> .


To study their differential activation properties , we linked the isolated effector domains to the GAL4 DNA-binding domain .

To study their differential activation properties , we linked the isolated <protein>effector domains</protein> to the <protein>GAL4 DNA-binding domain</protein> .


We have shown that both activating regions of Oct-2a , isolated from their natural context , can activate transcription as promoter factors .

We have shown that both activating regions of <protein>Oct-2a</protein> , isolated from their natural context , can activate transcription as <protein>promoter factors</protein> .


In contrast to the C-terminus , activation by the N-terminal domain is dependent on a yet unidentified factor ( s ) binding to the simian virus 40 enhancer .

In contrast to the <protein>C-terminus</protein> , activation by the <protein>N-terminal domain</protein> is dependent on a yet unidentified factor ( s ) binding to the <dna>simian virus 40 enhancer</dna> .


The results obtained by duplication of activation domains or their mixed combination suggest that the domains are functionally independent .

The results obtained by duplication of <protein>activation domains</protein> or their mixed combination suggest that the domains are functionally independent .


However , activation from a remote position could only be achieved with the C-terminus of Oct-2a in B cells .

However , activation from a remote position could only be achieved with the <protein>C-terminus</protein> of <protein>Oct-2a</protein> in <cell_type>B cells</cell_type> .


In lymphoid cells , higher activation levels were observed , suggesting that distinct B-cell-specific cofactors in concert with the effector domains of Oct-2a might be involved in mediating transcription from proximal and remote positions .

In <cell_type>lymphoid cells</cell_type> , higher activation levels were observed , suggesting that distinct <protein>B-cell-specific cofactors</protein> in concert with the <protein>effector domains</protein> of <protein>Oct-2a</protein> might be involved in mediating transcription from proximal and remote positions .


Furthermore , we identified a repression domain at the N-terminus of Oct-2a .

Furthermore , we identified a repression domain at the N-terminus of <protein>Oct-2a</protein> .


When transferred to a potent activator , transcriptional stimulation was inhibited efficiently .


These results underscore the modular structure of Oct-2a with separable domains for activation and repression and suggest that Oct-2a might have complex regulatory functions in vivo .

These results underscore the modular structure of <protein>Oct-2a</protein> with separable domains for activation and repression and suggest that <protein>Oct-2a</protein> might have complex regulatory functions in vivo .


Nonopsonic phagocytosis of Pseudomonas aeruginosa by macrophages and polymorphonuclear leukocytes requires the presence of the bacterial flagellum .

Nonopsonic phagocytosis of Pseudomonas aeruginosa by <cell_type>macrophages</cell_type> and <cell_type>polymorphonuclear leukocytes</cell_type> requires the presence of the bacterial flagellum .


Whereas the mechanism of nonopsonic phagocytosis of Pseudomonas aeruginosa has been described , the bacterial ligands required are poorly understood .


To identify the requisite bacterial ligands , studies with isogenic mutants of P. aeruginosa PAK lacking pili , flagella , and the RpoN sigma factor were undertaken .

To identify the requisite bacterial ligands , studies with isogenic mutants of P. aeruginosa PAK lacking pili , flagella , and the <protein>RpoN sigma factor</protein> were undertaken .


The RpoN mutant , lacking pili , flagella , and nonpilus adhesins , bound poorly and was resistant to ingestion by both macrophages and neutrophils .

The RpoN mutant , lacking pili , flagella , and <protein>nonpilus adhesins</protein> , bound poorly and was resistant to ingestion by both <cell_type>macrophages</cell_type> and <cell_type>neutrophils</cell_type> .


Pili were not absolutely required for binding or phagocytosis of P. aeruginosa .


The presence of a flagellum was not required for binding of P. aeruginosa to macrophages but was critical for the subsequent internalization of the bacterium , suggesting that this factor or a surface ligand associated with its assembly was responsible for stimulation of nonopsonic phagocytosis .

The presence of a flagellum was not required for binding of P. aeruginosa to <cell_type>macrophages</cell_type> but was critical for the subsequent internalization of the bacterium , suggesting that this factor or a surface ligand associated with its assembly was responsible for stimulation of nonopsonic phagocytosis .


Identification of essential GATA and Ets binding motifs within the promoter of the platelet glycoprotein Ib alpha gene .

Identification of essential <dna>GATA</dna> and <dna>Ets binding motifs</dna> within the promoter of the <dna>platelet glycoprotein Ib alpha gene</dna> .


Platelet glycoprotein ( GP ) Ib-IX-V is a multisubunit adhesion receptor that supports platelet attachment to thrombogenic surfaces at sites of vascular injury .

<protein>Platelet glycoprotein</protein> ( <protein>GP</protein> ) <protein>Ib-IX-V</protein> is a <protein>multisubunit adhesion receptor</protein> that supports platelet attachment to thrombogenic surfaces at sites of vascular injury .


The congenital absence of the receptor results in a bleeding disorder associated with `` giant '' platelets , a condition linking the expression of the complex to platelet morphogenesis .

The congenital absence of the receptor results in a bleeding disorder associated with <cell_type>`` giant '' platelets</cell_type> , a condition linking the expression of the complex to platelet morphogenesis .


To understand better the expression of the GP Ib-IX-V complex , studies were undertaken to define the essential genetic elements supporting the expression of the alpha-subunit of the complex ( GP Ib alpha ) .

To understand better the expression of the <protein>GP Ib-IX-V complex</protein> , studies were undertaken to define the essential <dna>genetic elements</dna> supporting the expression of the <protein>alpha-subunit</protein> of the complex ( <protein>GP Ib alpha</protein> ) .


GP Ib alpha promoter activity was evaluated by transfection of human erythroleukemia cells with reporter plasmids coding for the enzyme , luciferase .

<dna>GP Ib alpha promoter</dna> activity was evaluated by transfection of <cell_type>human erythroleukemia cells</cell_type> with reporter plasmids coding for the <protein>enzyme</protein> , <protein>luciferase</protein> .


Studies were initiated with a fragment extending 2 , 738 nucleotides 5 ' to the transcription start site and lead to the identification of 253 nucleotides retaining full promoter activity in human erythroleukemia cells .

Studies were initiated with a fragment extending 2 , 738 nucleotides 5 ' to the <dna>transcription start site</dna> and lead to the identification of 253 nucleotides retaining full promoter activity in <cell_type>human erythroleukemia cells</cell_type> .


In cells of nonhematopoietic lineage , human endothelial and HeLa cells , the GP Ib alpha promoter activity was no greater than background levels obtained with promoterless constructs .

In cells of <cell_type>nonhematopoietic lineage</cell_type> , <cell_type>human endothelial</cell_type> and <cell_type>HeLa cells</cell_type> , the <dna>GP Ib alpha promoter</dna> activity was no greater than background levels obtained with <dna>promoterless constructs</dna> .


Gel shift assays and site-directed mutagenesis studies defined essential GATA and Ets binding motifs 93 and 150 nucleotides upstream of the transcription start site , a finding which further substantiates these elements as important determinants of megakaryocytic gene expression .

Gel shift assays and site-directed mutagenesis studies defined essential <dna>GATA</dna> and <dna>Ets binding motifs</dna> <dna>93 and 150 nucleotides upstream</dna> of the <dna>transcription start site</dna> , a finding which further substantiates these elements as important determinants of megakaryocytic gene expression .


The results define essential cis-acting elements responsible for the expression of GP Ib alpha and provide insights into molecular events coinciding with the release of normal platelets into the bloodstream .

The results define essential <dna>cis-acting elements</dna> responsible for the expression of <protein>GP Ib alpha</protein> and provide insights into molecular events coinciding with the release of <cell_type>normal platelets</cell_type> into the bloodstream .


CD30 ligation induces nuclear factor-kappa B activation in human T cell lines .

<protein>CD30</protein> ligation induces <protein>nuclear factor-kappa B</protein> activation in <cell_line>human T cell lines</cell_line> .


CD30 is a recently described member of the tumor necrosis factor/nerve growth factor receptor superfamily .

<protein>CD30</protein> is a recently described member of the <protein>tumor necrosis factor/nerve growth factor receptor superfamily</protein> .


In this report , we show that following incubation of L540 cells ( Hodgkin 's disease-derived , T cell-like , CD30+ cells ) with the agonistic anti-CD30 monoclonal antibodies ( mAb ) M44 and M67 , two nuclear factor ( NF ) -kappa B DNA binding activities were induced in nuclear extracts , as determined in gel retardation assays .

In this report , we show that following incubation of <cell_line>L540 cells</cell_line> ( <cell_line>Hodgkin 's disease-derived , T cell-like , CD30+ cells</cell_line> ) with the <protein>agonistic anti-CD30 monoclonal antibodies</protein> ( <protein>mAb</protein> ) <protein>M44</protein> and <protein>M67</protein> , two <protein>nuclear factor ( NF ) -kappa B</protein> DNA binding activities were induced in nuclear extracts , as determined in gel retardation assays .


The effect of the mAb towards NF-kappa B activation was rapid , as it occurred within 20 min , and was sustained for up to 6 h .

The effect of the <protein>mAb</protein> towards <protein>NF-kappa B</protein> activation was rapid , as it occurred within 20 min , and was sustained for up to 6 h .


By comparison , an isotype-matched antibody had no effect on NF-kappa B activation .

By comparison , an <protein>isotype-matched antibody</protein> had no effect on <protein>NF-kappa B</protein> activation .


Moreover , in human T helper ( Th ) clones functionally characterized as being of the type 0 , type 1 and type 2 ( 28 % , < 1 % und 93 % CD30+ , respectively ) , the extent of CD30 -mediated NF-kappa B activation correlated with the proportion of CD30+ cells .

Moreover , in <cell_line>human T helper ( Th ) clones</cell_line> functionally characterized as being of the type 0 , type 1 and type 2 ( 28 % , < 1 % und 93 % CD30+ , respectively ) , the extent of <protein>CD30</protein> -mediated <protein>NF-kappa B</protein> activation correlated with the proportion of <cell_line>CD30+ cells</cell_line> .


In all cell lines investigated , the NF-kappa B complexes induced following CD30 engagement were shown to contain p50 NF-kappa B1 , p65 RelA , and possibly other transcription factors .

In all cell lines investigated , the <protein>NF-kappa B complexes</protein> induced following <protein>CD30</protein> engagement were shown to contain <protein>p50 NF-kappa B1</protein> , <protein>p65 RelA</protein> , and possibly other <protein>transcription factors</protein> .


Collectively , our results demonstrate that nuclear translocation and activation of NF-kappa B rank among the short-term cellular responses elicited following CD30 ligation .

Collectively , our results demonstrate that nuclear translocation and activation of <protein>NF-kappa B</protein> rank among the short-term cellular responses elicited following <protein>CD30</protein> ligation .


Pathogenesis of atherosclerosis .


The earliest lesion in the development of an atherosclerotic plaque is the fatty streak .


This chronic inflammatory reaction results from a sequence of events that begins with the trapping of low density lipoprotein ( LDL ) in the subendothelial space of the artery wall .


The trapped LDL is seeded with oxidative species released by the overlying endothelium , and lipid oxidation is initiated within the LDL particle. Some of the lipids that result lead to the activation of NFkB-like transcription factors that cause the expression of genes whose protein products mediate monocyte binding , monocyte chemotaxis into the subendothelial space , and conversion into macrophages .

The trapped LDL is seeded with oxidative species released by the overlying endothelium , and lipid oxidation is initiated within the LDL particle. Some of the lipids that result lead to the activation of <protein>NFkB-like transcription factors</protein> that cause the expression of genes whose <protein>protein products</protein> mediate monocyte binding , <cell_type>monocyte</cell_type> chemotaxis into the subendothelial space , and conversion into <cell_type>macrophages</cell_type> .


At least 1 major gene modulates the oxidation of LDL lipids and/or the biologic response to these lipids .

At least 1 <dna>major gene</dna> modulates the oxidation of LDL lipids and/or the biologic response to these lipids .


The inverse relation between high density lipoprotein ( HDL ) and atherosclerotic events may in part be due to enzymes associated with HDL that destroy the biologically active lipids generated in LDL .

The inverse relation between high density lipoprotein ( HDL ) and atherosclerotic events may in part be due to <protein>enzymes</protein> associated with HDL that destroy the biologically active lipids generated in LDL .


Estrogen and progesterone receptors in vernal keratoconjunctivitis .

<protein>Estrogen</protein> and <protein>progesterone receptors</protein> in vernal keratoconjunctivitis .


PURPOSE : Sex-related influences have been implicated in the pathogenesis of vernal keratoconjunctivitis ( VKC ) , an allergic eosinophilic disease .


METHODS : The authors evaluated tarsal and bulbar conjunctival biopsies from seven patients with severe and symptomatic VKC for the presence of estrogen and progesterone receptors by using monoclonal antibodies with a peroxidase-antiperoxidase technique .

METHODS : The authors evaluated tarsal and bulbar conjunctival biopsies from seven patients with severe and symptomatic VKC for the presence of <protein>estrogen</protein> and <protein>progesterone receptors</protein> by using <protein>monoclonal antibodies</protein> with a peroxidase-antiperoxidase technique .


RESULTS : Both the epithelium and subepithelium of the tarsal and bulbar conjunctiva of patients with VKC , but not those of four nonatopic control subjects , showed intense positive staining for estrogen and progesterone receptors .

RESULTS : Both the epithelium and subepithelium of the tarsal and bulbar conjunctiva of patients with VKC , but not those of four nonatopic control subjects , showed intense positive staining for estrogen and <protein>progesterone receptors</protein> .


Immunofluorescence colocalization of both estrogen and progesterone receptors with eosinophil cationic protein showed that approximately 70 % of positive cells were eosinophils .

Immunofluorescence colocalization of both <protein>estrogen</protein> and <protein>progesterone receptors</protein> with <protein>eosinophil cationic protein</protein> showed that approximately 70 % of <cell_type>positive cells</cell_type> were <cell_type>eosinophils</cell_type> .


CONCLUSIONS : Sexual hormones , through their receptors , may influence the activity of eosinophils in patients with VKC .

CONCLUSIONS : Sexual hormones , through their <protein>receptors</protein> , may influence the activity of <cell_type>eosinophils</cell_type> in patients with VKC .


CIITA activates the expression of MHC class II genes in mouse T cells .

<protein>CIITA</protein> activates the expression of <dna>MHC class II genes</dna> in mouse T cells .


It has long been a puzzle that MHC class II molecules are expressed in human T cells after activation but not in mouse T cells ; this expression is believed to play a role in the cell mediated immune response .

It has long been a puzzle that <protein>MHC class II molecules</protein> are expressed in <cell_type>human T cells</cell_type> after activation but not in <cell_type>mouse T cells</cell_type> ; this expression is believed to play a role in the cell mediated immune response .


Recently the MHC class II transactivator ( CIITA ) has been reported to be a major regulatory factor for both the constitutive and IFN inducible expression of MHC class II genes .

Recently the <protein>MHC class II transactivator</protein> ( <protein>CIITA</protein> ) has been reported to be a <protein>major regulatory factor</protein> for both the constitutive and IFN inducible expression of <dna>MHC class II genes</dna> .


Here we show that human T cells expressing MHC class II have CIITA transcripts while MHC class II-negative human T cells and mouse T cells do not .

Here we show that <cell_type>human T cells</cell_type> expressing MHC class II have <protein>CIITA</protein> transcripts while MHC class II-negative <cell_type>human T cells</cell_type> and <cell_type>mouse T cells</cell_type> do not .


The expression of MHC class II genes in mouse T cells can be reconstituted upon transfection with the human CIITA cDNA .

The expression of <dna>MHC class II genes</dna> in <cell_type>mouse T cells</cell_type> can be reconstituted upon transfection with the <dna>human CIITA cDNA</dna> .


These data indicate that the expression of CIITA explains the expression or lack of expression of MHC class II in human and mouse T cells respectively .

These data indicate that the expression of <protein>CIITA</protein> explains the expression or lack of expression of <protein>MHC class II</protein> in <cell_type>human</cell_type> and <cell_type>mouse T cells</cell_type> respectively .


Anti-Ro ( SSA ) autoantibodies are associated with T cell receptor beta genes in systemic lupus erythematosus patients .

<protein>Anti-Ro ( SSA ) autoantibodies</protein> are associated with <dna>T cell receptor beta genes</dna> in systemic lupus erythematosus patients .


Several of the heterogeneous clinical manifestations of systemic lupus erythematosus have been associated with specific autoantibodies .

Several of the heterogeneous clinical manifestations of systemic lupus erythematosus have been associated with specific <protein>autoantibodies</protein> .


Associations between HLA class II antigens and autoantibodies to the ribonucleoproteins Ro ( SSA ) and La ( SSB ) have been reported in these patients .

Associations between <protein>HLA class II antigens</protein> and <protein>autoantibodies</protein> to the <protein>ribonucleoproteins</protein> <protein>Ro ( SSA )</protein> and <protein>La ( SSB )</protein> have been reported in these patients .


Because HLA class II molecules present antigen to T cell receptors ( TCRs ) , we have searched for a TCR gene associated with the production of anti-Ro ( SSA ) antibodies .

Because <protein>HLA class II molecules</protein> present antigen to <protein>T cell receptors</protein> ( <protein>TCRs</protein> ) , we have searched for a <dna>TCR gene</dna> associated with the production of <protein>anti-Ro ( SSA ) antibodies</protein> .


A pair of restriction fragment length polymorphisms ( RFLPs ) , one of which hybridizes to the TCR constant region C beta 1 and the other to the C beta 2 gene , has been identified , suggesting these may be genotypic markers for an extended region of the TCR beta locus .

A pair of <dna>restriction fragment length polymorphisms</dna> ( <dna>RFLPs</dna> ) , one of which hybridizes to the <dna>TCR constant region C beta 1</dna> and the other to the <dna>C beta 2 gene</dna> , has been identified , suggesting these may be genotypic markers for an extended region of the <dna>TCR beta locus</dna> .


This RFLP pair occurs in 76 % of patients with Ro ( SSA ) precipitins , 84 % of anti- Ro ( SSA ) -positive patients lacking La ( SSB ) precipitins , but only 41 % of the patients lacking both precipitins ( P = 0.0004 ) .

This <dna>RFLP pair</dna> occurs in 76 % of patients with <protein>Ro ( SSA )</protein> precipitins , 84 % of anti- <protein>Ro ( SSA )</protein> -positive patients lacking <protein>La ( SSB )</protein> <protein>precipitins</protein> , but only 41 % of the patients lacking both <protein>precipitins</protein> ( P = 0.0004 ) .


This disproportionate occurrence in a subset of lupus patients indicates that these RFLPs are not disease susceptibility markers , but rather are important markers for TCR genes whose products are involved in the production of anti-Ro ( SSA ) antibodies .

This disproportionate occurrence in a subset of lupus patients indicates that these <dna>RFLPs</dna> are not disease susceptibility markers , but rather are important markers for <dna>TCR genes</dna> whose products are involved in the production of <protein>anti-Ro ( SSA ) antibodies</protein> .


The majority of patients who have these RFLPs and HLA class II antigens previously associated with the anti- Ro ( SSA ) response make this antibody , suggesting that interactions between products of these loci occur in response to Ro ( SSA ) .

The majority of patients who have these <dna>RFLPs</dna> and <protein>HLA class II antigens</protein> previously associated with the anti- <protein>Ro ( SSA )</protein> response make this antibody , suggesting that interactions between products of these <dna>loci occur</dna> in response to <protein>Ro ( SSA )</protein> .


Inhibition of NF-AT -dependent transcription by NF-kappa B : implications for differential gene expression in T helper cell subsets .

Inhibition of <protein>NF-AT</protein> -dependent transcription by <protein>NF-kappa B</protein> : implications for differential gene expression in <cell_type>T helper cell subsets</cell_type> .


Activation of individual CD4+ T cells results in differential lymphokine expression : interleukin 2 ( IL-2 ) is preferentially produced by T helper type 1 ( TH1 ) cells , which are involved in cell-mediated immune responses , whereas IL-4 is synthesized by TH2 cells , which are essential for humoral immunity .

Activation of individual <cell_line>CD4+ T cells</cell_line> results in differential <protein>lymphokine</protein> expression : <protein>interleukin 2</protein> ( <protein>IL-2</protein> ) is preferentially produced by <cell_line>T helper type 1 ( TH1 ) cells</cell_line> , which are involved in cell-mediated immune responses , whereas <protein>IL-4</protein> is synthesized by <cell_line>TH2 cells</cell_line> , which are essential for humoral immunity .


The Ca ( 2+ ) -dependent factor NF-ATp plays a key role in the inducible transcription of both these lymphokine genes .

The <protein>Ca ( 2+ ) -dependent factor</protein> <protein>NF-ATp</protein> plays a key role in the inducible transcription of both these <dna>lymphokine genes</dna> .


However , while IL2 expression requires the contribution of Ca ( 2+ ) - and protein kinase C -dependent signals , we report that activation of human IL4 transcription through the Ca ( 2+ ) -dependent pathway is diminished by protein kinase C stimulation in Jurkat T cells .

However , while <protein>IL2</protein> expression requires the contribution of Ca ( 2+ ) - and <protein>protein kinase C</protein> -dependent signals , we report that activation of <protein>human IL4</protein> transcription through the Ca ( 2+ ) -dependent pathway is diminished by <protein>protein kinase C</protein> stimulation in <cell_line>Jurkat T cells</cell_line> .


This phenomenon is due to mutually exclusive binding of NF-ATp and NF-kappa B to the P sequence , an element located 69 bp upstream of the IL4 transcription initiation site .

This phenomenon is due to mutually exclusive binding of <protein>NF-ATp</protein> and <protein>NF-kappa B</protein> to the <dna>P sequence</dna> , an element located <dna>69 bp upstream</dna> of the <dna>IL4 transcription initiation site</dna> .


Human IL4 promoter -mediated transcription is downregulated in Jurkat cells stimulated with the NF-kappa B-activating cytokine tumor necrosis factor alpha and suppressed in RelA-overexpressing cells .

<dna>Human IL4 promoter</dna> -mediated transcription is downregulated in <cell_line>Jurkat cells</cell_line> stimulated with the <protein>NF-kappa B-activating cytokine</protein> <protein>tumor necrosis factor alpha</protein> and suppressed in <cell_line>RelA-overexpressing cells</cell_line> .


In contrast , protein kinase C stimulation or RelA overexpression does not affect the activity of a human IL4 promoter containing a mouse P sequence , which is a higher-affinity site for NF-ATp and a lower-affinity site for RelA .

In contrast , <protein>protein kinase C</protein> stimulation or <protein>RelA</protein> overexpression does not affect the activity of a <dna>human IL4 promoter</dna> containing a <dna>mouse P sequence</dna> , which is a <dna>higher-affinity site</dna> for <protein>NF-ATp</protein> and a <dna>lower-affinity site</dna> for <protein>RelA</protein> .


Thus , competition between two general transcriptional activators , RelA and NF-ATp , mediates the inhibitory effect of protein kinase C stimulation on IL4 expression and may contribute to differential gene expression in TH cells .

Thus , competition between two general <protein>transcriptional activators</protein> , <protein>RelA</protein> and <protein>NF-ATp</protein> , mediates the inhibitory effect of <protein>protein kinase C</protein> stimulation on <protein>IL4</protein> expression and may contribute to differential gene expression in <cell_line>TH cells</cell_line> .


Regulation of the balance of cytokine production and the signal transducer and activator of transcription ( STAT ) transcription factor activity by cytokines and inflammatory synovial fluids .

Regulation of the balance of <protein>cytokine</protein> production and the signal transducer and activator of transcription ( <protein>STAT</protein> ) transcription factor activity by <protein>cytokines</protein> and inflammatory synovial fluids .


The balance between type 1 and 2 T helper cell cytokine production plays an important role in several animal models of autoimmunity , and skewed patterns of cytokine expression have been described in human inflammatory diseases .

The balance between type 1 and 2 T helper cell cytokine production plays an important role in several animal models of autoimmunity , and skewed patterns of <protein>cytokine</protein> expression have been described in human inflammatory diseases .


Many cytokines activate signal transducer and activation of transcription ( STAT ) transcription factors , which , in turn , activate transcription of inflammatory effector genes .

Many <protein>cytokines</protein> activate <protein>signal transducer and activation of transcription ( STAT ) transcription factors</protein> , which , in turn , activate transcription of inflammatory effector genes .


We used mononuclear cell priming cultures and inflammatory synovial fluids ( SFs ) derived from arthritis patients to examine the regulation of cytokine production and STAT activity by an inflammatory synovial microenvironment .

We used <cell_line>mononuclear cell priming cultures</cell_line> and <cell_type>inflammatory synovial fluids</cell_type> ( <cell_type>SFs</cell_type> ) derived from arthritis patients to examine the regulation of <protein>cytokine</protein> production and <protein>STAT</protein> activity by an inflammatory synovial microenvironment .


Exposure to SFs during priming resulted in an 81 % inhibition of interferon ( IFN ) -gamma , but not interleukin ( IL ) 4 , production by effector cells generated in priming cultures .

Exposure to <cell_type>SFs</cell_type> during priming resulted in an 81 % inhibition of <protein>interferon ( IFN ) -gamma</protein> , but not <protein>interleukin ( IL ) 4</protein> , production by <cell_type>effector cells</cell_type> generated in priming cultures .


SF suppression was mediated by IL-4 and IL-10 and inhibition of IL-12 expression , and it was reversed in a dominant fashion by exogenous IL-12 .

SF suppression was mediated by <protein>IL-4</protein> and <protein>IL-10</protein> and inhibition of <protein>IL-12</protein> expression , and it was reversed in a dominant fashion by exogenous IL-12 .


SFs blocked the sustained activity of transcription factor Stat1 , but not Stat3 , during the priming period , and Stat1 activity was differentially regulated by cytokines in parallel with their positive or negative regulation of IFN-gamma production .

<cell_type>SFs</cell_type> blocked the sustained activity of <protein>transcription factor Stat1</protein> , but not <protein>Stat3</protein> , during the priming period , and <protein>Stat1</protein> activity was differentially regulated by <protein>cytokines</protein> in parallel with their positive or negative regulation of <protein>IFN-gamma</protein> production .


Active Stat3 , but not Stat1 , was detected in cells from inflamed joints .

Active <protein>Stat3</protein> , but not <protein>Stat1</protein> , was detected in cells from inflamed joints .


These results suggest a role for altered balance of Stat1 and Stat3 transcriptional activity in the regulation of T cell differentiation and in the pathogenesis of inflammatory synovitis .

These results suggest a role for altered balance of Stat1 and Stat3 transcriptional activity in the regulation of <cell_type>T cell</cell_type> differentiation and in the pathogenesis of inflammatory synovitis .


Triggering of complement receptors CR1 ( CD35 ) and CR3 ( CD11b/CD18 ) induces nuclear translocation of NF-kappa B ( p50/p65 ) in human monocytes and enhances viral replication in HIV-infected monocytic cells .

Triggering of <protein>complement receptors</protein> <protein>CR1</protein> ( <protein>CD35</protein> ) and <protein>CR3</protein> ( <protein>CD11b/CD18</protein> ) induces nuclear translocation of <protein>NF-kappa B</protein> ( <protein>p50/p65</protein> ) in <cell_type>human monocytes</cell_type> and enhances viral replication in <cell_type>HIV-infected monocytic cells</cell_type> .


Monocyte/macrophages may harbor HIV in a nonproductive fashion for prolonged periods of time .

<cell_type>Monocyte/macrophages</cell_type> may harbor HIV in a nonproductive fashion for prolonged periods of time .


Viral gene expression may be reactivated by stimulation of the cells with LPS or cytokines such as TNF-alpha in vitro .

Viral gene expression may be reactivated by stimulation of the cells with LPS or <protein>cytokines</protein> such as <protein>TNF-alpha</protein> in vitro .


The effect of LPS and TNF-alpha is mediated by their ability to induce nuclear translocation of the DNA-binding heterodimer NF-kappa B ( p50/p65 ) , which binds to a specific sequence in the HIV-long terminal repeat .

The effect of LPS and <protein>TNF-alpha</protein> is mediated by their ability to induce nuclear translocation of the <protein>DNA-binding heterodimer</protein> <protein>NF-kappa B</protein> ( <protein>p50/p65</protein> ) , which binds to a specific sequence in the <dna>HIV-long terminal repeat</dna> .


The present study demonstrates that triggering of complement receptors CR1 ( CD35 ) and CR3 ( CD11b/CD18 ) enhances viral replication in HIV-infected human monocytic cells .

The present study demonstrates that triggering of <protein>complement receptors</protein> <protein>CR1</protein> ( <protein>CD35</protein> ) and <protein>CR3</protein> ( <protein>CD11b/CD18</protein> ) enhances viral replication in <cell_line>HIV-infected human monocytic cells</cell_line> .


Monocytic cell lines and normal peripheral blood monocytes were infected with HIV-1 in vitro and cultured in the presence or absence of F ( ab ' ) 2 fragments of monoclonal anti-CR1 or anti-CR3 Abs or with C3 fragments .

<cell_line>Monocytic cell lines</cell_line> and normal peripheral blood monocytes were infected with HIV-1 in vitro and cultured in the presence or absence of <protein>F ( ab ' ) 2 fragments</protein> of <protein>monoclonal anti-CR1</protein> or <protein>anti-CR3 Abs</protein> or with <protein>C3 fragments</protein> .


Stimulation of CR1 or CR3 induces a two- to fourfold increase in the amount of cell-associated and released p24 Ag in cell cultures that was equivalent to that observed in control cultures triggered with LPS .

Stimulation of <protein>CR1</protein> or <protein>CR3</protein> induces a two- to fourfold increase in the amount of cell-associated and released <protein>p24 Ag</protein> in <cell_line>cell cultures</cell_line> that was equivalent to that observed in <cell_line>control cultures</cell_line> triggered with LPS .


We further observed that stimulation of CR1 or CR3 induces the nuclear translocation of NF-kappa B p50/p65 in infected cells .

We further observed that stimulation of <protein>CR1</protein> or <protein>CR3</protein> induces the nuclear translocation of <protein>NF-kappa B</protein> <protein>p50/p65</protein> in <cell_type>infected cells</cell_type> .


Translocation of NF-kappa B p50/p65 was also observed following stimulation of CR1 or CR3 of uninfected peripheral blood monocytes from HIV-seronegative donors .

Translocation of <protein>NF-kappa B</protein> <protein>p50/p65</protein> was also observed following stimulation of <protein>CR1</protein> or <protein>CR3</protein> of <cell_type>uninfected peripheral blood monocytes</cell_type> from HIV-seronegative donors .


The amount of protein translocated was similar to that observed when cells were stimulated with rhTNF-alpha .

The amount of protein translocated was similar to that observed when cells were stimulated with <protein>rhTNF-alpha</protein> .


TNF-alpha did not mediate the translocation of NF-kappa B p50/p65 induced by triggering of complement receptors .

<protein>TNF-alpha</protein> did not mediate the translocation of <protein>NF-kappa B</protein> <protein>p50/p65</protein> induced by triggering of <protein>complement receptors</protein> .


Taken together , these observations suggest that HIV gene expression may be activated in infected monocytes through interaction of the cells with complement-opsonized particles and that enhanced viral replication is associated with C3 receptor -mediated nuclear translocation of the NF-kappa B complex .

Taken together , these observations suggest that HIV gene expression may be activated in <cell_type>infected monocytes</cell_type> through interaction of the cells with <protein>complement-opsonized particles</protein> and that enhanced viral replication is associated with <protein>C3 receptor</protein> -mediated nuclear translocation of the <protein>NF-kappa B complex</protein> .


Attenuation of gamma interferon -induced tyrosine phosphorylation in mononuclear phagocytes infected with Leishmania donovani : selective inhibition of signaling through Janus kinases and Stat1 .

Attenuation of <protein>gamma interferon</protein> -induced tyrosine phosphorylation in <cell_type>mononuclear phagocytes</cell_type> infected with Leishmania donovani : selective inhibition of signaling through <protein>Janus kinases</protein> and <protein>Stat1</protein> .


The induction of gene transcription in response to gamma interferon is impaired in mononuclear phagocytes infected with Leishmania donovani , and the mechanisms involved are not fully understood .

The induction of gene transcription in response to <protein>gamma interferon</protein> is impaired in <cell_type>mononuclear phagocytes</cell_type> infected with Leishmania donovani , and the mechanisms involved are not fully understood .


The changes in gene expression brought about by gamma interferon are thought to involve transient increases in the activities of cellular protein tyrosine kinases , including the Janus kinases Jak1 and Jak2 , leading to tyrosine phosphorylation of the transcription factor Stat1 .

The changes in gene expression brought about by <protein>gamma interferon</protein> are thought to involve transient increases in the activities of <protein>cellular protein tyrosine kinases</protein> , including the <protein>Janus kinases</protein> <protein>Jak1</protein> and <protein>Jak2</protein> , leading to tyrosine phosphorylation of the <protein>transcription factor Stat1</protein> .


To investigate the mechanisms accounting for the impaired responses to gamma interferon , a model system for examining overall changes in protein tyrosine phosphorylation , activation of Jak1 and Jak2 and phosphorylation of Stat1 was developed in phorbol 12-myristate 13-acetate-differentiated U-937 cells .

To investigate the mechanisms accounting for the impaired responses to <protein>gamma interferon</protein> , a model system for examining overall changes in protein tyrosine phosphorylation , activation of <protein>Jak1</protein> and <protein>Jak2</protein> and phosphorylation of <protein>Stat1</protein> was developed in <cell_line>phorbol 12-myristate 13-acetate-differentiated U-937 cells</cell_line> .


Analysis of whole-cell lysates by antiphosphotyrosine immunoblotting showed that incubation with gamma interferon brought about specific increases in phosphotyrosine labeling of several proteins .

Analysis of whole-cell lysates by antiphosphotyrosine immunoblotting showed that incubation with <protein>gamma interferon</protein> brought about specific increases in phosphotyrosine labeling of several proteins .


Increased labeling of these proteins occurred to similar extents in control cells and in cells that had been infected with L. donovani for 16 h .

Increased labeling of these proteins occurred to similar extents in <cell_type>control cells</cell_type> and in cells that had been infected with L. donovani for 16 h .


Jak1 , Jak2 , and Stat1 were immunoprecipitated from control and interferon-treated cells , and tyrosine phosphorylation of these proteins , detected by antiphosphotyrosine immunoblotting was used to measured their activation .

<protein>Jak1</protein> , <protein>Jak2</protein> , and <protein>Stat1</protein> were immunoprecipitated from control and <cell_line>interferon-treated cells</cell_line> , and tyrosine phosphorylation of these proteins , detected by antiphosphotyrosine immunoblotting was used to measured their activation .


Tyrosine phosphorylation of Jak1 , Jak2 , and Stat1 increased markedly , in a dose-dependent manner , in U-937 cells incubated with gamma interferon .

Tyrosine phosphorylation of <protein>Jak1</protein> , <protein>Jak2</protein> , and <protein>Stat1</protein> increased markedly , in a dose-dependent manner , in <cell_line>U-937 cells</cell_line> incubated with <protein>gamma interferon</protein> .


In contrast , in cells infected with L. donovani , tyrosine phosphorylation of Jak1 , Jak2 , and Stat1 was markedly impaired .

In contrast , in cells infected with L. donovani , tyrosine phosphorylation of <protein>Jak1</protein> , <protein>Jak2</protein> , and <protein>Stat1</protein> was markedly impaired .


This effect was dependent upon the duration of exposure to L. donovani and was maximal and complete at 16 h .


Results similar to those observed with U-937 cells were also obtained with human peripheral blood monocytes .

Results similar to those observed with <cell_line>U-937 cells</cell_line> were also obtained with <cell_type>human peripheral blood monocytes</cell_type> .


These findings indicate that infection of human mononuclear phagocytes with L. donovani leads to impaired gamma interferon -mediated tyrosine phosphorylation and selective effects on the Jak- Stat1 pathway .

These findings indicate that infection of <cell_type>human mononuclear phagocytes</cell_type> with L. donovani leads to impaired <protein>gamma interferon</protein> -mediated tyrosine phosphorylation and selective effects on the Jak- <protein>Stat1</protein> pathway .


Unresponsiveness to gamma interferon for activation of this pathway may explain impaired transcriptional responses in leishmania-infected cells .

Unresponsiveness to <protein>gamma interferon</protein> for activation of this pathway may explain impaired transcriptional responses in <cell_type>leishmania-infected cells</cell_type> .


Mutually exclusive interaction of a novel matrix attachment region binding protein and the NF-muNR enhancer repressor .

Mutually exclusive interaction of a novel <protein>matrix attachment region binding protein</protein> and the <protein>NF-muNR enhancer repressor</protein> .


Implications for regulation of immunoglobulin heavy chain expression .

Implications for regulation of <protein>immunoglobulin heavy chain</protein> expression .


The immunoglobulin heavy chain ( IgH ) intronic enhancer stimulates transcription from functional promoters in B lymphocytes but not other cell types .

The <protein>immunoglobulin heavy chain</protein> ( <protein>IgH</protein> ) intronic enhancer stimulates transcription from functional promoters in B lymphocytes but not other cell types .


The observation that binding sites for the nuclear factor-mu negative regulator ( NF-muNR ) enhancer repressor overlap nuclear matrix attachment regions ( MARs ) in this enhancer has lead to the hypothesis that the cell type specificity of the enhancer might be controlled by regulating nuclear matrix attachment ( Scheuermann , R. H. , and Chen , U. ( 1989 ) Genes & Dev. 3 , 1255-1266 ) .

The observation that binding sites for the <protein>nuclear factor-mu negative regulator ( NF-muNR ) enhancer repressor overlap nuclear matrix attachment regions</protein> ( <protein>MARs</protein> ) in this <dna>enhancer</dna> has lead to the hypothesis that the cell type specificity of the <dna>enhancer</dna> might be controlled by regulating nuclear matrix attachment ( Scheuermann , R. H. , and Chen , U. ( 1989 ) Genes & Dev. 3 , 1255-1266 ) .


To understand the role of MARs in IgH enhancer regulation , we have identified a novel MAR-binding protein , MAR-BP1 , from soluble nuclear matrix preparations based on its ability to bind to the MARs associated with the IgH enhancer .

To understand the role of <protein>MARs</protein> in <dna>IgH enhancer</dna> regulation , we have identified a novel <protein>MAR-binding protein</protein> , <protein>MAR-BP1</protein> , from soluble nuclear matrix preparations based on its ability to bind to the <protein>MARs</protein> associated with the <dna>IgH enhancer</dna> .


Purified MAR-BP1 migrates as a 33-kDa protein , and it can be found in nuclear matrix preparations from a number of different types of lymphoid cell lines .

Purified <protein>MAR-BP1</protein> migrates as a <protein>33-kDa protein</protein> , and it can be found in nuclear matrix preparations from a number of different types of <cell_line>lymphoid cell lines</cell_line> .


Although specific binding sites have been difficult to localize by chemical or enzymatic footprinting procedures , NF-muNR binding sites are critical for efficient MAR-BP1 binding .

Although specific binding sites have been difficult to localize by chemical or enzymatic footprinting procedures , <protein>NF-muNR</protein> binding sites are critical for efficient <protein>MAR-BP1</protein> binding .


Indeed , binding of the IgH enhancer to either intact nuclear matrix preparations or to MAR-BP1 is mutually exclusive to NF-muNR binding .

Indeed , binding of the <dna>IgH enhancer</dna> to either intact nuclear matrix preparations or to <protein>MAR-BP1</protein> is mutually exclusive to <protein>NF-muNR</protein> binding .


These results are consistent with a model for cell-type specific regulation in which binding of the NF-muNR repressor to the IgH enhancer prevents nuclear matrix attachment in inappropriate cells by interfering with MAR-BP1 /enhancer interaction .

These results are consistent with a model for cell-type specific regulation in which binding of the <protein>NF-muNR repressor</protein> to the <dna>IgH enhancer</dna> prevents nuclear matrix attachment in inappropriate cells by interfering with <protein>MAR-BP1</protein> <dna>/enhancer</dna> interaction .


PU.1 ( Spi-1 ) and C/EBP alpha regulate expression of the granulocyte-macrophage colony-stimulating factor receptor alpha gene .

<protein>PU.1</protein> ( <protein>Spi-1</protein> ) and <protein>C/EBP alpha</protein> regulate expression of the <dna>granulocyte-macrophage colony-stimulating factor receptor alpha gene</dna> .


Growth factor receptors play an important role in hematopoiesis .

<protein>Growth factor receptors</protein> play an important role in hematopoiesis .


In order to further understand the mechanisms directing the expression of these key regulators of hematopoiesis , we initiated a study investigating the transcription factors activating the expression of the granulocyte-macrophage colony-stimulating factor ( GM-CSF ) receptor alpha gene .

In order to further understand the mechanisms directing the expression of these key regulators of hematopoiesis , we initiated a study investigating the <protein>transcription factors</protein> activating the expression of the <dna>granulocyte-macrophage colony-stimulating factor ( GM-CSF ) receptor alpha gene</dna> .


Here , we demonstrate that the human GM-CSF receptor alpha promoter directs reporter gene activity in a tissue-specific fashion in myelomonocytic cells , which correlates with its expression pattern as analyzed by reverse transcription PCR .

Here , we demonstrate that the <dna>human GM-CSF receptor alpha promoter</dna> directs <dna>reporter gene</dna> activity in a tissue-specific fashion in <cell_type>myelomonocytic cells</cell_type> , which correlates with its expression pattern as analyzed by reverse transcription PCR .


The GM-CSF receptor alpha promoter contains an important functional site between positions -53 and -41 as identified by deletion analysis of reporter constructs .

The <dna>GM-CSF receptor alpha promoter</dna> contains an important functional site between positions -53 and -41 as identified by deletion analysis of <dna>reporter constructs</dna> .


We show that the myeloid and B cell transcription factor PU.1 binds specifically to this site .

We show that the myeloid and <protein>B cell transcription factor</protein> <protein>PU.1</protein> binds specifically to this site .


Furthermore , we demonstrate that a CCAAT site located upstream of the PU.1 site between positions -70 and -54 is involved in positive-negative regulation of the GM-CSF receptor alpha promoter activity .

Furthermore , we demonstrate that a <dna>CCAAT site</dna> located upstream of the <dna>PU.1 site</dna> between positions -70 and -54 is involved in positive-negative regulation of the <dna>GM-CSF receptor alpha promoter</dna> activity .


C/EBP alpha is the major CCAAT/enhancer-binding protein ( C/EBP ) form binding to this site in nuclear extracts of U937 cells .

<protein>C/EBP alpha</protein> is the major <protein>CCAAT/enhancer-binding protein</protein> ( <protein>C/EBP</protein> ) form binding to this site in nuclear extracts of <cell_line>U937 cells</cell_line> .


Point mutations of either the PU.1 site or the C/EBP site that abolish the binding of the respective factors result in a significant decrease of GM-CSF receptor alpha promoter activity in myelomonocytic cells only .

Point mutations of either the <dna>PU.1 site</dna> or the <dna>C/EBP site</dna> that abolish the binding of the respective factors result in a significant decrease of <dna>GM-CSF receptor alpha promoter</dna> activity in <cell_type>myelomonocytic cells</cell_type> only .


Furthermore , we demonstrate that in myeloid and B cell extracts , PU.1 forms a novel , specific , more slowly migrating complex ( PU-SF ) when binding the GM-CSF receptor alpha promoter PU.1 site .

Furthermore , we demonstrate that in myeloid and B cell extracts , <protein>PU.1</protein> forms a novel , specific , more slowly migrating complex ( <protein>PU-SF</protein> ) when binding the <protein>GM-CSF receptor alpha</protein> promoter <dna>PU.1 site</dna> .


This is the first demonstration of a specific interaction with PU.1 on a myeloid PU.1 binding site .

This is the first demonstration of a specific interaction with <protein>PU.1</protein> on a <dna>myeloid PU.1 binding site</dna> .


The novel complex is distinct from that described previously as binding to B cell enhancer sites and can be formed by addition of PU.1 to extracts from certain nonmyeloid cell types which do not express PU.1 , including T cells and epithelial cells , but not from erythroid cells .

The novel complex is distinct from that described previously as binding to <dna>B cell enhancer sites</dna> and can be formed by addition of <protein>PU.1</protein> to extracts from certain <cell_type>nonmyeloid cell types</cell_type> which do not express <protein>PU.1</protein> , including <cell_type>T cells</cell_type> and <cell_type>epithelial cells</cell_type> , but not from <cell_type>erythroid cells</cell_type> .


Furthermore , we demonstrate that the PU-SF complex binds to PU.1 sites found on a number of myeloid promoters , and its formation requires an intact PU.1 site adjacent to a single-stranded region .

Furthermore , we demonstrate that the <protein>PU-SF complex</protein> binds to <protein>PU.1</protein> sites found on a number of <dna>myeloid promoters</dna> , and its formation requires an intact <dna>PU.1 site</dna> adjacent to a single-stranded region .


Expression of PU.1 in nonmyeloid cells can activate the GM-CSF receptor alpha promoter .

Expression of <protein>PU.1</protein> in <cell_type>nonmyeloid cells</cell_type> can activate the <dna>GM-CSF receptor alpha promoter</dna> .


Deletion of the amino-terminal region of PU.1 results in a failure to form the PU-SF complex and in a concomitant loss of transactivation , suggesting that formation of the PU-SF complex is of functional importance for the activity of the GM-CSF receptor alpha promoter .

Deletion of the <protein>amino-terminal region</protein> of <protein>PU.1</protein> results in a failure to form the <protein>PU-SF</protein> complex and in a concomitant loss of transactivation , suggesting that formation of the <protein>PU-SF complex</protein> is of functional importance for the activity of the <dna>GM-CSF receptor alpha promoter</dna> .


Finally , we demonstrate that C/EBP alpha can also active the GM-CSF receptor alpha promoter in nonmyeloid cells .

Finally , we demonstrate that <protein>C/EBP alpha</protein> can also active the <dna>GM-CSF receptor alpha promoter</dna> in <cell_type>nonmyeloid cells</cell_type> .


( ABSTRACT TRUNCATED AT 400 WORDS )


HMG-I binds to GATA motifs : implications for an HPFH syndrome .

<protein>HMG-I</protein> binds to <dna>GATA motifs</dna> : implications for an HPFH syndrome .


We have examined binding of the nuclear protein HMG-I to the human gamma-globin promoter .

We have examined binding of the <protein>nuclear protein</protein> <protein>HMG-I</protein> to the <dna>human gamma-globin promoter</dna> .


We find that HMG-I binds preferentially to the more 3 ' of a pair of GATA motifs in the gamma-globin promoter ; this paired motif is bound by the erythroid factor GATA-1 .

We find that <protein>HMG-I</protein> binds preferentially to the more 3 ' of a pair of <dna>GATA motifs</dna> in the <dna>gamma-globin promoter</dna> ; this paired motif is bound by the <protein>erythroid factor GATA-1</protein> .


A naturally occurring mutation ( -175 T-C ) in the area bound by HMG-I results in overexpression of gamma-globin in adult red blood cells ( HPFH ) and up-regulation of the gamma-globin promoter in in vitro expression assays ; HMG-I does not bind to this mutant sequence .

A naturally occurring mutation ( -175 T-C ) in the area bound by <protein>HMG-I</protein> results in overexpression of <protein>gamma-globin</protein> in <cell_type>adult red blood cells</cell_type> ( <cell_type>HPFH</cell_type> ) and up-regulation of the <dna>gamma-globin promoter</dna> in in vitro expression assays ; <protein>HMG-I</protein> does not bind to this <dna>mutant sequence</dna> .


A survey of GATA motifs from other globin cis-elements demonstrates HMG-I binding to most of them .

A survey of <dna>GATA motifs</dna> from other <dna>globin cis-elements</dna> demonstrates <protein>HMG-I</protein> binding to most of them .


These findings implicate HMG-I in the HPFH phenotype ; we speculate that it may participate in the formation of multiprotein complexes that regulate globin gene expression .

These findings implicate <protein>HMG-I</protein> in the <cell_type>HPFH</cell_type> phenotype ; we speculate that it may participate in the formation of <protein>multiprotein complexes</protein> that regulate globin gene expression .


Constitutive activation of different Jak tyrosine kinases in human T cell leukemia virus type 1 ( HTLV-1 ) tax protein or virus-transformed cells .

Constitutive activation of different <protein>Jak tyrosine kinases</protein> in <protein>human T cell leukemia virus type 1 ( HTLV-1 ) tax protein</protein> or <cell_line>virus-transformed cells</cell_line> .


HTLV-1 infection causes an adult T cell leukemia in humans .


The viral encoded protein tax , is thought to play an important role in oncogenesis .

The <protein>viral encoded protein</protein> <protein>tax</protein> , is thought to play an important role in oncogenesis .


Our previous data obtained from a tax transgenic mouse model revealed that tax transforms mouse fibroblasts but not thymocytes , despite comparable levels of tax expression in both tissues .

Our previous data obtained from a <protein>tax</protein> transgenic mouse model revealed that <protein>tax</protein> transforms <cell_type>mouse fibroblasts</cell_type> but not <cell_type>thymocytes</cell_type> , despite comparable levels of <protein>tax</protein> expression in both tissues .


Constitutive tyrosine phosphorylation of a 130-kD protein ( s ) was observed in the tax transformed fibroblast B line and in HTLV-1 transformed human lymphoid lines , but not in thymocytes from Thy-tax transgenic mice .

Constitutive tyrosine phosphorylation of a <protein>130-kD protein</protein> ( s ) was observed in the <protein>tax</protein> <cell_line>transformed fibroblast B line</cell_line> and in <cell_line>HTLV-1 transformed human lymphoid lines</cell_line> , but not in <cell_type>thymocytes</cell_type> from Thy-tax transgenic mice .


Phosphotyrosine immunoprecipitation followed by Western blot analysis with a set of Jak kinase specific antibodies , identified p130 as Jak2 in the tax transformed mouse fibroblastic cell line and Jak3 in HTLV-1 transformed human T cell lines .

Phosphotyrosine immunoprecipitation followed by Western blot analysis with a set of <protein>Jak kinase specific antibodies</protein> , identified p130 as <protein>Jak2</protein> in the <protein>tax</protein> transformed <cell_line>mouse fibroblastic cell line</cell_line> and <protein>Jak3</protein> in <cell_line>HTLV-1 transformed human T cell lines</cell_line> .


Phosphorylation of Jak2 in tax transformed cells resulted from high expression of IL-6 .

Phosphorylation of <protein>Jak2</protein> in <cell_line>tax transformed cells</cell_line> resulted from high expression of IL-6 .


Tyrosine phosphorylation of this protein could also be induced in Balb/c3T3 cells using a supernatant from the B line , which was associated with induction of cell proliferation .

Tyrosine phosphorylation of this protein could also be induced in <cell_line>Balb/c3T3 cells</cell_line> using a supernatant from the <cell_line>B line</cell_line> , which was associated with induction of cell proliferation .


Both phosphorylation and proliferation were inhibited by IL-6 neutralizing antibodies .

Both phosphorylation and proliferation were inhibited by <protein>IL-6 neutralizing antibodies</protein> .


Constitutive phosphorylation of Jak kinases may facilitate tumor growth in both HTLV-1 infected human T cells and the transgenic mouse model .

Constitutive phosphorylation of <protein>Jak kinases</protein> may facilitate tumor growth in both <cell_type>HTLV-1 infected human T cells</cell_type> and the transgenic mouse model .


Regulation of c-jun mRNA expression by hydroxyurea in human K562 cells during erythroid differentiation [ published erratum appears in Biochim Biophys Acta 1995 Dec 27 ; 1264 ( 3 ) : 409 ]

Regulation of <rna>c-jun mRNA</rna> expression by hydroxyurea in <cell_line>human K562 cells</cell_line> during erythroid differentiation [ published erratum appears in Biochim Biophys Acta 1995 Dec 27 ; 1264 ( 3 ) : 409 ]


Hydroxyurea ( HU ) is an antitumor agent which also induces hemoglobinization during erythroid differentiation .


In addition , HU stimulates the synthesis of fetal hemoglobin in sickle cell anemia patients .

In addition , HU stimulates the synthesis of <protein>fetal hemoglobin</protein> in sickle cell anemia patients .


To further understand its mechanism of action , we investigated the effects of HU on regulation of c-jun expression prior to the onset of erythroid differentiation of K562 cells .

To further understand its mechanism of action , we investigated the effects of HU on regulation of <dna>c-jun</dna> expression prior to the onset of erythroid differentiation of <cell_line>K562 cells</cell_line> .


HU induced a dose-dependent stimulation of c-jun synthesis .

HU induced a dose-dependent stimulation of <dna>c-jun</dna> synthesis .


The levels of c-jun mRNA was elevated 4 to 7.5-fold by HU within 2 h .

The levels of <rna>c-jun mRNA</rna> was elevated 4 to 7.5-fold by HU within 2 h .


This was followed by a gradual decline to the basal level by 24 h .


Both nuclear run-on and actinomycin D pulse experiments strongly indicate that HU regulates c-jun mRNA expression by increasing the rate of synthesis as well as stabilizing the c-jun mRNA .

Both nuclear run-on and actinomycin D pulse experiments strongly indicate that HU regulates <rna>c-jun mRNA</rna> expression by increasing the rate of synthesis as well as stabilizing the <rna>c-jun mRNA</rna> .


In addition , the level of jun protein was elevated by 2 to 5-fold within 4 h in HU treated cells .

In addition , the level of <protein>jun protein</protein> was elevated by 2 to 5-fold within 4 h in <cell_line>HU treated cells</cell_line> .


Furthermore , concentrations of HU below 250 microM slightly increased the 5X AP-1 /CAT activity .

Furthermore , concentrations of HU below 250 microM slightly increased the 5X <protein>AP-1</protein> <protein>/CAT</protein> activity .


These results strongly suggest that HU induces both transcriptional and post-transcription regulation of c-jun during erythroid differentiation .

These results strongly suggest that HU induces both transcriptional and post-transcription regulation of <dna>c-jun</dna> during erythroid differentiation .


In vivo and in vitro effects of glucocorticoids on lymphocyte proliferation in man : relationship to glucocorticoid receptors .

In vivo and in vitro effects of glucocorticoids on lymphocyte proliferation in man : relationship to <protein>glucocorticoid receptors</protein> .


Interrelations between the hypothalamic-pituitary-adrenal system ( HPA ) and the immune system represent a well-documented biological phenomenon .


While in vitro administration of glucocorticoids may inhibit concanavalin A ( Con A ) - and phytohemagglutinin ( PHA ) -induced T-cell proliferation , pokeweed mitogen ( PWM ) -driven B-cell mitogenesis is relatively resistant to glucocorticoids .

While in vitro administration of glucocorticoids may inhibit concanavalin A ( Con A ) - and phytohemagglutinin ( PHA ) -induced T-cell proliferation , <protein>pokeweed mitogen</protein> ( <protein>PWM</protein> ) -driven B-cell mitogenesis is relatively resistant to glucocorticoids .


To further explore the link between the HPA and the immune system in relation to glucocorticoid receptor function , dose-response curves were obtained for Con A -and PHA -induced T-cell mitogenesis , PWM -generated B-cell mitogenesis and spontaneous lymphocyte proliferation in 13 healthy controls .

To further explore the link between the HPA and the immune system in relation to <protein>glucocorticoid receptor</protein> function , dose-response curves were obtained for Con A -and <protein>PHA</protein> -induced T-cell mitogenesis , <protein>PWM</protein> -generated B-cell mitogenesis and spontaneous lymphocyte proliferation in 13 healthy controls .


Glucocorticoid effects were assessed in vivo by depletion of endogenous glucocorticoids after oral administration of 1.5 g metyrapone ( MET ) and subsequent glucocorticoid replacement , and in vitro by incubation of the cells with different doses of dexamethasone ( DEX ) .


There was a significant decrease in PWM -induced B-cell mitogenesis and a more pronounced effect of DEX administered in vitro on spontaneous lymphocyte proliferation after MET treatment when compared with the DEX plus MET pretreated condition in vivo .

There was a significant decrease in <protein>PWM</protein> -induced B-cell mitogenesis and a more pronounced effect of DEX administered in vitro on spontaneous lymphocyte proliferation after MET treatment when compared with the DEX plus MET pretreated condition in vivo .


These data suggest that the inhibition of spontaneous lymphocyte proliferation by glucocorticoids in vitro is related to glucocorticoid receptor function .

These data suggest that the inhibition of spontaneous lymphocyte proliferation by glucocorticoids in vitro is related to <protein>glucocorticoid receptor</protein> function .


The decrease in PWM -generated B-cell proliferation following cortisol depletion by MET may be seen in connection with impaired glucocorticoid-mediated induction of interleukin-1 receptor synthesis .

The decrease in <protein>PWM</protein> -generated B-cell proliferation following cortisol depletion by MET may be seen in connection with impaired glucocorticoid-mediated induction of <protein>interleukin-1 receptor</protein> synthesis .