Although IFN-gamma alone does not induce ISG expression , IFN-gamma pretreatment markedly increases and hastens ISG expression and transcriptional induction .

Although <protein>IFN-gamma</protein> alone does not induce ISG expression , <protein>IFN-gamma</protein> pretreatment markedly increases and hastens ISG expression and transcriptional induction .


Nuclear extracts assayed for the presence of ISRE binding factors by electrophoretic mobility shift assays show that ISGF3 is induced by IFN-alpha within 6 h from undetectable basal levels in untreated U937 cells .

Nuclear extracts assayed for the presence of <protein>ISRE binding factors</protein> by electrophoretic mobility shift assays show that <protein>ISGF3</protein> is induced by <protein>IFN-alpha</protein> within 6 h from undetectable basal levels in untreated <cell_line>U937 cells</cell_line> .


Activation of ISGF3 alpha , the latent component of ISGF3 , occurs rapidly .

Activation of <protein>ISGF3 alpha</protein> , the latent component of <protein>ISGF3</protein> , occurs rapidly .


However , the increase in ISGF3 activity ultimately correlates with the accumulation of ISGF3 gamma induced by IFN-alpha or IFN-gamma .

However , the increase in <protein>ISGF3</protein> activity ultimately correlates with the accumulation of <protein>ISGF3 gamma</protein> induced by <protein>IFN-alpha</protein> or <protein>IFN-gamma</protein> .


( ABSTRACT TRUNCATED AT 250 WORDS )


Single point estimation of glucocorticoid receptors in lymphocytes of normal subjects and of children under long term glucocorticoid treatment .

Single point estimation of <protein>glucocorticoid receptors</protein> in <cell_type>lymphocytes</cell_type> of normal subjects and of children under long term glucocorticoid treatment .


A single point assay of glucocorticoid receptors ( GR ) in human lymphocytes based on the measurement of specific dexamethasone binding has been developed and compared with a common multi-point Scatchard analysis .

A single point assay of <protein>glucocorticoid receptors</protein> ( <protein>GR</protein> ) in human <cell_type>lymphocytes</cell_type> based on the measurement of specific dexamethasone binding has been developed and compared with a common multi-point Scatchard analysis .


The assay conditions-concentration of the ligand 20 nmol/l , incubation time 2 h and the cell count 2-6 mil .


cells/tube in the assay volume 0.25 ml were found to be optimal .


An attempt was also undertaken to use a cell harvester for the separation of cells from unbound ligand .


Though specifically bound dexamethasone measured by whole-cell assay and that using cell harvester correlated well , almost by one order lower values obtained with the latter method render it non-applicable for receptor quantitation .


The results from 9 healthy volunteers ( average GR concentration 7131 +/- 1256 sites/cell ) correlated excellently with those obtained by the Scatchard analysis .

The results from 9 healthy volunteers ( average <protein>GR</protein> concentration 7131 +/- 1256 sites/cell ) correlated excellently with those obtained by the Scatchard analysis .


The single point assay has been also applied for determination of GH in 10 children treated with large doses of prednisone .


The average values from healthy volunteers did not differ significantly from those found in these children , though much broader range was found in patients .


Glucocorticoid receptor and inhibition of 3-O-methyl-D-glucose uptake by glucocorticoids in peripheral blood leukocytes from normal humans : correlation between receptor level and hormone effect in vitro .

<protein>Glucocorticoid receptor</protein> and inhibition of 3-O-methyl-D-glucose uptake by glucocorticoids in <cell_type>peripheral blood leukocytes</cell_type> from normal humans : correlation between receptor level and hormone effect in vitro .


We have measured the glucocorticoid receptor concentration in mononuclear and polymorphonuclear leukocytes , both of which were isolated from peripheral blood from ten healthy male volunteers .

We have measured the <protein>glucocorticoid receptor</protein> concentration in <cell_type>mononuclear</cell_type> and <cell_type>polymorphonuclear leukocytes</cell_type> , both of which were isolated from peripheral blood from ten healthy male volunteers .


In parallel , the inhibitory effect of dexamethasone on 3-O-methyl-D-glucose uptake was assayed in the corresponding mononuclear leukocytes .

In parallel , the inhibitory effect of dexamethasone on 3-O-methyl-D-glucose uptake was assayed in the corresponding <cell_type>mononuclear leukocytes</cell_type> .


The glucocorticoid receptor levels in mononuclear leukocytes correlated with those in polymorphonuclear leukocytes , and there was a linear relationship between the cellular glucocorticoid receptor levels and glucocorticoid-mediated inhibition of the uptake of 3-O-methyl-D-glucose in mononuclear leukocytes .

The <protein>glucocorticoid receptor</protein> levels in <cell_type>mononuclear leukocytes</cell_type> correlated with those in <cell_type>polymorphonuclear leukocytes</cell_type> , and there was a linear relationship between the cellular <protein>glucocorticoid receptor</protein> levels and glucocorticoid-mediated inhibition of the uptake of 3-O-methyl-D-glucose in <cell_type>mononuclear leukocytes</cell_type> .


When mononuclear leukocytes were incubated in the presence of 8-bromo-cAMP , cellular glucocorticoid receptor levels increased and a more pronounced inhibitory effect of dexamethasone was observed on the transport of 3-O-methyl-D-glucose .

When <cell_type>mononuclear leukocytes</cell_type> were incubated in the presence of 8-bromo-cAMP , cellular <protein>glucocorticoid receptor</protein> levels increased and a more pronounced inhibitory effect of dexamethasone was observed on the transport of 3-O-methyl-D-glucose .


We conclude that the cellular glucocorticoid receptor levels in peripheral blood leukocytes reflect in vitro responsiveness to glucocorticoids in mononuclear leukocytes from healthy males , and that the individual responsiveness may alter upon changes in the cellular levels of glucocorticoid receptor .

We conclude that the cellular <protein>glucocorticoid receptor</protein> levels in <cell_type>peripheral blood leukocytes</cell_type> reflect in vitro responsiveness to glucocorticoids in <cell_type>mononuclear leukocytes</cell_type> from healthy males , and that the individual responsiveness may alter upon changes in the cellular levels of <protein>glucocorticoid receptor</protein> .


Transcription factor activation and functional stimulation of human monocytes .

<protein>Transcription factor</protein> activation and functional stimulation of <cell_type>human monocytes</cell_type> .


Activation of expression of genes encoding transcription factors : c-fos and c-jun and formation of AP1 transcriptional complex in human monocytes was investigated .

Activation of expression of <dna>genes</dna> encoding <protein>transcription factors</protein> : <dna>c-fos</dna> and <dna>c-jun</dna> and formation of <protein>AP1 transcriptional complex</protein> in <cell_type>human monocytes</cell_type> was investigated .


It was found that lipopolysaccharide induced strongly both c-fos and c-jun expression as well as AP1 formation .

It was found that lipopolysaccharide induced strongly both <dna>c-fos</dna> and <dna>c-jun</dna> expression as well as <protein>AP1</protein> formation .


Interferon gamma activated strongly c-fos and weakly c-jun and AP1 .

<protein>Interferon gamma</protein> activated strongly <dna>c-fos</dna> and weakly <dna>c-jun</dna> and <protein>AP1</protein> .


Tumor necrosis factor induced slightly c-fos and had almost no effect on c-jun and AP1 .

<protein>Tumor necrosis factor</protein> induced slightly <dna>c-fos</dna> and had almost no effect on <dna>c-jun</dna> and <protein>AP1</protein> .


The data suggest that differences in functional responses elicited in monocytes by all three factors may be dependent on different routes on nuclear signalling employed by the factors .

The data suggest that differences in functional responses elicited in <cell_type>monocytes</cell_type> by all three <protein>factors</protein> may be dependent on different routes on nuclear signalling employed by the <protein>factors</protein> .


Regulation of interleukin-1 beta production by glucocorticoids in human monocytes : the mechanism of action depends on the activation signal .

Regulation of <protein>interleukin-1 beta</protein> production by glucocorticoids in <cell_type>human monocytes</cell_type> : the mechanism of action depends on the activation signal .


Glucocorticoids are known to downregulate interleukin-1 beta production in monocytic cells by two different mechanims : direct inhibition of the gene transcription and destabilization of the preformed interleukin-1 beta mRNA .

Glucocorticoids are known to downregulate <protein>interleukin-1 beta</protein> production in <cell_type>monocytic cells</cell_type> by two different mechanims : direct inhibition of the gene transcription and destabilization of the preformed <rna>interleukin-1 beta mRNA</rna> .


Now we have examined the effect of the nature of the monocyte activating signal on these two inhibitory mechanims .


When human monocytes were preincubated with dexamethasone for 1 hour and then stimulated either with bacterial lipopolysaccharide or phorbol myristate , it was found that dexamethasone inhibited the lipopolysaccharide-induced interleukin-1 beta protein production , but the phorbol myristate-induced production was increased 3-10 fold .

When <cell_type>human monocytes</cell_type> were preincubated with dexamethasone for 1 hour and then stimulated either with bacterial lipopolysaccharide or phorbol myristate , it was found that dexamethasone inhibited the lipopolysaccharide-induced <protein>interleukin-1 beta</protein> protein production , but the phorbol myristate-induced production was increased 3-10 fold .


This difference was also seen at the mRNA level .


When dexamethasone was added to the cultures 3 hours after the stimulators , it clearly decreased the interleukin-1 beta mRNA levels regardless of the stimulator used ( although the effect was clearly weaker on the PMA-induced mRNA ) .

When dexamethasone was added to the cultures 3 hours after the stimulators , it clearly decreased the <rna>interleukin-1 beta mRNA</rna> levels regardless of the stimulator used ( although the effect was clearly weaker on the <rna>PMA-induced mRNA</rna> ) .


Thus these data suggest that the phorbol myristate-induced signal ( prolonged protein kinase C activation ? ) can not be inhibited by prior incubation with dexamethasone and it also protects the induced mRNA for the degradative action of dexamethasone .

Thus these data suggest that the phorbol myristate-induced signal ( prolonged <protein>protein kinase C</protein> activation ? ) can not be inhibited by prior incubation with dexamethasone and it also protects the <rna>induced mRNA</rna> for the degradative action of dexamethasone .


Nuclear transcription factors that bind to elements of the IL-2 promoter .

<protein>Nuclear transcription factors</protein> that bind to elements of the <dna>IL-2 promoter</dna> .


Induction requirements in primary human T cells .

Induction requirements in <cell_type>primary human T cells</cell_type> .


Prior studies have identified several elements that contribute to the activity of the IL-2 promoter in the stimulated T cell line , Jurkat .

Prior studies have identified several elements that contribute to the activity of the <dna>IL-2 promoter</dna> in the stimulated <cell_line>T cell line</cell_line> , <cell_line>Jurkat</cell_line> .


The sites and their corresponding nuclear binding factors include : NF-kappa B , AP-1 , AP-3 , OCT-1 , and NF-AT .

The sites and their corresponding nuclear binding factors include : <protein>NF-kappa B</protein> , <protein>AP-1</protein> , <protein>AP-3</protein> , <protein>OCT-1</protein> , and <protein>NF-AT</protein> .


The latter `` nuclear factor for activated T cells `` likely contributes to the tissue specificity of IL-2 gene expression .

The latter `` <protein>nuclear factor for activated T cells</protein> `` likely contributes to the tissue specificity of <dna>IL-2 gene</dna> expression .


Using electrophoretic mobility shift assays , we have studied these transcription factors in primary T cells from human blood to verify their presence in a physiologic setting and to identify the signals that stimulate factor activity .

Using electrophoretic mobility shift assays , we have studied these <protein>transcription factors</protein> in <cell_type>primary T cells</cell_type> from human blood to verify their presence in a physiologic setting and to identify the signals that stimulate factor activity .


All factors are induced in the nuclei of T cells upon activation with mitogens but not with exogenous IL-2 growth factor .

All factors are induced in the nuclei of <cell_type>T cells</cell_type> upon activation with <protein>mitogens</protein> but not with <protein>exogenous IL-2 growth factor</protein> .


However , the signaling requirements and sensitivity to protein synthesis inhibitors differ considerably .


Only the activities for NF-AT and AP-1 sites require two signals for optimal induction , i.e. , PMA plus either lectin or antibody to the CD3 or CD28 surface molecules .

Only the activities for <dna>NF-AT and AP-1 sites</dna> require two signals for optimal induction , i.e. , PMA plus either <protein>lectin</protein> or <protein>antibody</protein> to the <protein>CD3</protein> or <protein>CD28 surface molecules</protein> .


Other factors are induced by lectin , antibody , and/or PMA alone .

Other factors are induced by <protein>lectin</protein> , <protein>antibody</protein> , and/or PMA alone .


After appropriate stimulation , both NF-AT and AP-1 are peculiarly sensitive to the protein synthesis inhibitor anisomycin .

After appropriate stimulation , both <protein>NF-AT</protein> and <protein>AP-1</protein> are peculiarly sensitive to the protein synthesis inhibitor anisomycin .


Our data correlate the activity of NF-AT and AP-1 in gel shift assays with the two signals requirements for IL-2 gene expression .

Our data correlate the activity of <protein>NF-AT</protein> and <protein>AP-1</protein> in gel shift assays with the two signals requirements for <dna>IL-2 gene</dna> expression .


NF-kappa B activity in T cells stably expressing the Tax protein of human T cell lymphotropic virus type I .

<protein>NF-kappa B</protein> activity in <cell_type>T cells</cell_type> stably expressing the <protein>Tax protein</protein> of human T cell lymphotropic virus type I .


The effect of constitutive Tax expression on the interaction of NF-kappa B with its recognition sequence and on NF-kappa B -dependent gene expression was examined in T lymphoid Jurkat cell lines ( 19D and 9J ) stably transformed with a Tax expression vector .

The effect of constitutive <protein>Tax</protein> expression on the interaction of <protein>NF-kappa B</protein> with its <dna>recognition sequence</dna> and on <protein>NF-kappa B</protein> -dependent gene expression was examined in <cell_line>T lymphoid Jurkat cell lines</cell_line> ( <cell_line>19D</cell_line> and <cell_line>9J</cell_line> ) stably transformed with a <dna>Tax expression vector</dna> .


Tax expressing T cell lines contained a constitutive level of NF-kappa B binding activity , detectable by mobility shift assay and uv cross-linking using a palindromic NF-kappa B probe homologous to the interferon beta PRDII site .

<cell_line>Tax expressing T cell lines</cell_line> contained a constitutive level of <protein>NF-kappa B</protein> binding activity , detectable by mobility shift assay and uv cross-linking using a <dna>palindromic NF-kappa B probe</dna> homologous to the <dna>interferon beta PRDII site</dna> .


In Jurkat and NC2.10 induction with phorbol esters resulted in the appearance of new DNA binding proteins of 85 , 75 , and 54 kDa , whereas in Tax expressing cells the 85-kDa protein and a 92-kDa DNA binding protein were constitutively induced .

In Jurkat and NC2.10 induction with phorbol esters resulted in the appearance of new <protein>DNA binding proteins</protein> of <protein>85 , 75 , and 54 kDa</protein> , whereas in <cell_line>Tax expressing cells</cell_line> the <protein>85-kDa protein</protein> and a <protein>92-kDa DNA binding protein</protein> were constitutively induced .


Expression of Tax protein in 19D and 9J resulted in transcription of the endogenous NF-kappa B-dependent granulocyte-macrophage colony stimulating factor gene and increased basal level expression of transfected NF-kappa B-regulated promoters .

Expression of <protein>Tax protein</protein> in <cell_line>19D</cell_line> and <cell_line>9J</cell_line> resulted in transcription of the <dna>endogenous NF-kappa B-dependent granulocyte-macrophage colony stimulating factor gene</dna> and increased basal level expression of transfected <dna>NF-kappa B-regulated promoters</dna> .


Nonetheless transcription of both the endogenous and the transfected gene was inducible by PMA treatment .


Tax expression in Jurkat T cells may alter the stoichiometry of NF-kappa B DNA binding proteins and thus change the expression of NF-kappa B-regulated promoters .

<protein>Tax</protein> expression in <cell_line>Jurkat T cells</cell_line> may alter the stoichiometry of <protein>NF-kappa B</protein> <protein>DNA binding proteins</protein> and thus change the expression of <dna>NF-kappa B-regulated promoters</dna> .


A nuclear factor NF-GM2 that interacts with a regulatory region of the GM-CSF gene essential for its induction in responses to T-cell activation : purification from human T-cell leukemia line Jurkat cells and similarity to NF-kappa B .

A <protein>nuclear factor</protein> <protein>NF-GM2</protein> that interacts with a <dna>regulatory region</dna> of the <dna>GM-CSF gene</dna> essential for its induction in responses to T-cell activation : purification from human <cell_line>T-cell leukemia line Jurkat cells</cell_line> and similarity to <protein>NF-kappa B</protein> .


Activation of T cells by antigen , lectin , or a combination of phorbol-12-myristate acetate ( PMA ) and calcium ionophore ( A23187 ) leads to the induction of genes for a set of lymphokines , including granulocyte-macrophage colony-stimulating factor ( GM-CSF ) .

Activation of <cell_type>T cells</cell_type> by antigen , <protein>lectin</protein> , or a combination of phorbol-12-myristate acetate ( PMA ) and calcium ionophore ( A23187 ) leads to the induction of genes for a set of <protein>lymphokines</protein> , including <protein>granulocyte-macrophage colony-stimulating factor</protein> ( <protein>GM-CSF</protein> ) .


We demonstrated in earlier studies that the upstream region of the mouse GM-CSF promoter at positions between -95 and -73 is essential for transcriptional activation in response to PMA/A23187 .

We demonstrated in earlier studies that the <dna>upstream region</dna> of the <dna>mouse GM-CSF promoter</dna> at positions between <dna>-95 and -73</dna> is essential for transcriptional activation in response to PMA/A23187 .


This region contains two DNA-binding motifs , GM2 and GC-box .

This region contains two <dna>DNA-binding motifs</dna> , <dna>GM2</dna> and <dna>GC-box</dna> .


The GM2 sequence ( GGTAGTTCCC ) is recognized by an inducible factor NF-GM2 ; the other ( CCGCCC ) by constitutive factors A1 , A2 , and B .

The <dna>GM2 sequence</dna> ( GGTAGTTCCC ) is recognized by an <protein>inducible factor NF-GM2</protein> ; the other ( CCGCCC ) by <protein>constitutive factors</protein> <protein>A1</protein> , <protein>A2</protein> , and <protein>B</protein> .


To elucidate the mechanism of GM-CSF gene activation , we have purified the inducible factor NF-GM2 from the nuclear extract of stimulated Jurkat cells on the basis of specific DNA-binding activity .

To elucidate the mechanism of <protein>GM-CSF</protein> gene activation , we have purified the <protein>inducible factor NF-GM2</protein> from the nuclear extract of stimulated <cell_line>Jurkat cells</cell_line> on the basis of specific DNA-binding activity .


The purified NF-GM2 consists of 50 ( p50 ) and 65 kDa ( p65 ) polypeptides and has a binding activity specific for both the GM-CSF and immunoglobulin kappa ( GGAAAGTCCC ) enhancers .

The purified <protein>NF-GM2</protein> consists of 50 ( <protein>p50</protein> ) and 65 kDa ( <protein>p65</protein> ) polypeptides and has a binding activity specific for both the <protein>GM-CSF</protein> and <dna>immunoglobulin kappa ( GGAAAGTCCC ) enhancers</dna> .


Electrophoretically purified p50 alone can form a protein-DNA complex , but in the mixture , p50 associates preferentially with p65 to form the NF-GM2 complex .

Electrophoretically purified <protein>p50</protein> alone can form a <protein>protein-DNA complex</protein> , but in the mixture , <protein>p50</protein> associates preferentially with <protein>p65</protein> to form the <protein>NF-GM2 complex</protein> .


In addition , p65 gave per se , with low affinity , a protein-DNA complex that migrated more slowly than native NF-GM2 complex .

In addition , <protein>p65</protein> gave per se , with low affinity , a <protein>protein-DNA complex</protein> that migrated more slowly than native <protein>NF-GM2 complex</protein> .


Furthermore , an antiserum against KBF1 ( identical to 50 kDa NF-kappa B protein ) reacted with the p50 of NF-GM2 , indicating that the NF-GM2 polypeptide can not be immunologically differentiated from the 50 kDa subunit of NF-kappa B .

Furthermore , an antiserum against <protein>KBF1</protein> ( identical to <protein>50 kDa NF-kappa B protein</protein> ) reacted with the <protein>p50</protein> of <protein>NF-GM2</protein> , indicating that the <protein>NF-GM2 polypeptide</protein> can not be immunologically differentiated from the <protein>50 kDa subunit</protein> of <protein>NF-kappa B</protein> .


The purified NF-GM2 activated in vitro transcription from the kappa B enhancer , while it failed to stimulate transcription from the GM-CSF promoter harboring the GM2 sequence .

The purified <protein>NF-GM2</protein> activated in vitro transcription from the <dna>kappa B enhancer</dna> , while it failed to stimulate transcription from the <dna>GM-CSF promoter</dna> harboring the GM2 sequence .


This suggests that the activation mechanism of the GM-CSF gene through the GM2/GC-box sequence is different from that of genes carrying the kappa B enhancer alone .

This suggests that the activation mechanism of the <dna>GM-CSF gene</dna> through the <dna>GM2/GC-box sequence</dna> is different from that of genes carrying the <dna>kappa B enhancer</dna> alone .


Characterization of an immediate-early gene induced in adherent monocytes that encodes I kappa B -like activity .

Characterization of an <dna>immediate-early gene</dna> induced in <cell_type>adherent monocytes</cell_type> that encodes <protein>I kappa B</protein> -like activity .


We have cloned a group of cDNAs representing mRNAs that are rapidly induced following adherence of human monocytes .

We have cloned a group of <dna>cDNAs</dna> representing <rna>mRNAs</rna> that are rapidly induced following adherence of <cell_type>human monocytes</cell_type> .


One of the induced transcripts ( MAD-3 ) encodes a protein of 317 amino acids with one domain containing five tandem repeats of the cdc10/ankyrin motif , which is 60 % similar ( 46 % identical ) to the ankyrin repeat region of the precursor of NF-kappa B/KBF1 p50 .

One of the induced transcripts ( <rna>MAD-3</rna> ) encodes a protein of 317 amino acids with one domain containing <protein>five tandem repeats</protein> of the <protein>cdc10/ankyrin motif</protein> , which is 60 % similar ( 46 % identical ) to the <protein>ankyrin repeat region</protein> of the precursor of <protein>NF-kappa B/KBF1 p50</protein> .


The C-terminus has a putative protein kinase C phosphorylation site .

The <protein>C-terminus</protein> has a putative <protein>protein kinase C phosphorylation site</protein> .


In vitro translated MAD-3 protein was found to specifically inhibit the DNA-binding activity of the p50/p65 NF-kappa B complex but not that of the p50/p50 KBF1 factor or of other DNA-binding proteins .

In vitro translated <protein>MAD-3 protein</protein> was found to specifically inhibit the DNA-binding activity of the <protein>p50/p65 NF-kappa B complex</protein> but not that of the <protein>p50/p50 KBF1 factor</protein> or of other <protein>DNA-binding proteins</protein> .


The MAD-3 cDNA encodes an I kappa B-like protein that is likely to be involved in regulation of transcriptional responses to NF-kappa B , including adhesion-dependent pathways of monocyte activation .

The <dna>MAD-3 cDNA</dna> encodes an <protein>I kappa B-like protein</protein> that is likely to be involved in regulation of transcriptional responses to <protein>NF-kappa B</protein> , including adhesion-dependent pathways of monocyte activation .


Cortivazol mediated induction of glucocorticoid receptor messenger ribonucleic acid in wild-type and dexamethasone-resistant human leukemic ( CEM ) cells .

Cortivazol mediated induction of <rna>glucocorticoid receptor messenger ribonucleic acid</rna> in <cell_line>wild-type</cell_line> and <cell_line>dexamethasone-resistant human leukemic ( CEM ) cells</cell_line> .


Cortivazol is a phenylpyrazolo glucocorticoid of high potency and unusual structure .


In both wild-type and highly dexamethasone ( dex ) -resistant clones of the human leukemic cell line CEM , exposure to cortivazol leads to cell death .

In both <cell_line>wild-type</cell_line> and highly <cell_line>dexamethasone ( dex ) -resistant clones</cell_line> of the <cell_line>human leukemic cell line CEM</cell_line> , exposure to cortivazol leads to cell death .


It has been shown recently that in wild-type CEM cells but not in a dex-resistant , glucocorticoid receptor ( GR ) -defective clone ICR-27 TK-3 , dex induces GR mRNA .

It has been shown recently that in <cell_line>wild-type CEM cells</cell_line> but not in a <cell_line>dex-resistant , glucocorticoid receptor ( GR ) -defective clone</cell_line> <cell_line>ICR-27 TK-3</cell_line> , dex induces <rna>GR mRNA</rna> .


To test the hypothesis that cortivazol acts in dex-resistant cells by making use of the residual GR found there , wild-type and dex-resistant clones were treated with various concentrations of cortivazol and induction of GR mRNA was studied .

To test the hypothesis that cortivazol acts in <cell_line>dex-resistant cells</cell_line> by making use of the residual <protein>GR</protein> found there , wild-type and <cell_line>dex-resistant clones</cell_line> were treated with various concentrations of cortivazol and induction of <rna>GR mRNA</rna> was studied .


Cortivazol significantly induced GR mRNA in the normal CEM-C7 as well as in two classes of dex-resistant clones , although the dex-resistant clones needed at least 10 times more cortivazol than the normal cells for significant GR mRNA induction .

Cortivazol significantly induced <rna>GR mRNA</rna> in the <cell_line>normal CEM-C7</cell_line> as well as in two classes of <cell_line>dex-resistant clones</cell_line> , although the <cell_line>dex-resistant clones</cell_line> needed at least 10 times more cortivazol than the <cell_type>normal cells</cell_type> for significant <rna>GR mRNA</rna> induction .


Increased levels of GR mRNA were noticed as early as 3 h after treatment .

Increased levels of <rna>GR mRNA</rna> were noticed as early as 3 h after treatment .


A general correlation between induction of GR mRNA and lysis of the normal and dex-resistant cells was found .

A general correlation between induction of <rna>GR mRNA</rna> and lysis of the normal and <cell_line>dex-resistant cells</cell_line> was found .


Positive induction of GR mRNA might be one of the earliest crucial steps in the lysis of normal and dex-resistant CEM cells , or might serve as a marker for the process .

Positive induction of <rna>GR mRNA</rna> might be one of the earliest crucial steps in the lysis of <cell_line>normal and dex-resistant CEM cells</cell_line> , or might serve as a marker for the process .


However , the lysis pathway in the dex-resistant cells is defective in that dex-resistant clones needed significantly more cortivazol than the normal cells for lysis of the cells .

However , the lysis pathway in the <cell_line>dex-resistant cells</cell_line> is defective in that <cell_line>dex-resistant clones</cell_line> needed significantly more cortivazol than the <cell_line>normal cells</cell_line> for lysis of the cells .


HIV enhancer activity perpetuated by NF-kappa B induction on infection of monocytes [ see comments ]

<dna>HIV enhancer</dna> activity perpetuated by <protein>NF-kappa B</protein> induction on infection of <cell_type>monocytes</cell_type> [ see comments ]


Permissiveness to replication of human immunodeficiency virus ( HIV ) differs in T lymphocytes and macrophages .

Permissiveness to replication of human immunodeficiency virus ( HIV ) differs in <cell_type>T lymphocytes</cell_type> and <cell_type>macrophages</cell_type> .


In T cells , HIV transcription is poorly detected in vivo .

In <cell_type>T cells</cell_type> , HIV transcription is poorly detected in vivo .


Cloned , normal T lymphocytes show very little , if any , basal activity of the HIV enhancer and low nuclear expression of NF-kappa B , a potent transcriptional activator of the HIV enhancer .

Cloned , normal <cell_type>T lymphocytes</cell_type> show very little , if any , basal activity of the <dna>HIV enhancer</dna> and low nuclear expression of <protein>NF-kappa B</protein> , a <protein>potent transcriptional activator</protein> of the <dna>HIV enhancer</dna> .


In contrast , fixed tissue macrophages express detectable HIV proteins , indicating permanent virus transcription .

In contrast , fixed tissue <cell_type>macrophages</cell_type> express detectable <protein>HIV proteins</protein> , indicating permanent virus transcription .


One explanation for the perpetuation of virus infection in macrophages could be sustained nuclear NF-kappa B expression .

One explanation for the perpetuation of virus infection in <cell_type>macrophages</cell_type> could be sustained nuclear <protein>NF-kappa B</protein> expression .


However , the U937 monocytic cell line , which is fully permissive to HIV replication , is known to express only low levels of nuclear NF-kappa B .

However , the <cell_line>U937 monocytic cell line</cell_line> , which is fully permissive to HIV replication , is known to express only low levels of <protein>nuclear NF-kappa B</protein> .


We show here that chronic HIV infection results in both induction of a nuclear factor with antigenic properties indistinguishable from those of NF-kappa B and permanently increased HIV enhancer activity .

We show here that chronic HIV infection results in both induction of a <protein>nuclear factor</protein> with antigenic properties indistinguishable from those of <protein>NF-kappa B</protein> and permanently increased <dna>HIV enhancer</dna> activity .


This phenomenon , which is independent of tumour necrosis factor , is associated with HIV replication , and is thus likely to explain at least in part the perpetuation of HIV infection in monocytes .

This phenomenon , which is independent of <protein>tumour necrosis factor</protein> , is associated with HIV replication , and is thus likely to explain at least in part the perpetuation of HIV infection in <cell_type>monocytes</cell_type> .


Isolation of a rel-related human cDNA that potentially encodes the 65-kD subunit of NF-kappa B [ published erratum appears in Science 1991 Oct 4 ; 254 ( 5028 ) : 11 ]

Isolation of a <dna>rel-related human cDNA</dna> that potentially encodes the <protein>65-kD subunit</protein> of <protein>NF-kappa B</protein> [ published erratum appears in Science 1991 Oct 4 ; 254 ( 5028 ) : 11 ]


A DNA probe that spanned a domain conserved among the proto-oncogene c-rel , the Drosophila morphogen dorsal , and the p50 DNA binding subunit of NF-kappa B was generated from Jurkat T cell complementary DNA with the polymerase chain reaction ( PCR ) and degenerate oligonucleotides .

A <dna>DNA probe</dna> that spanned a domain conserved among the <dna>proto-oncogene c-rel</dna> , the <protein>Drosophila morphogen dorsal</protein> , and the <protein>p50 DNA binding subunit</protein> of <protein>NF-kappa B</protein> was generated from <dna>Jurkat T cell complementary DNA</dna> with the polymerase chain reaction ( PCR ) and degenerate oligonucleotides .


This probe was used to identify a rel-related complementary DNA that hybridized to a 2.6-kilobase messenger RNA present in human T and B lymphocytes .

This probe was used to identify a <dna>rel-related complementary DNA</dna> that hybridized to a <rna>2.6-kilobase messenger RNA</rna> present in <cell_type>human T and B lymphocytes</cell_type> .


In vitro transcription and translation of the complementary DNA resulted in the synthesis of a protein with an apparent molecular size of 65 kilodaltons ( kD ) .

In vitro transcription and translation of the <dna>complementary DNA</dna> resulted in the synthesis of a protein with an apparent molecular size of <protein>65 kilodaltons</protein> ( kD ) .


The translated protein showed weak DNA binding with a specificity for the kappa B binding motif .


This protein-DNA complex comigrated with the complex obtained with the purified human p65 NF-kappa B subunit and binding was inhibited by I kappa B-alpha and -beta proteins .

This <protein>protein-DNA complex</protein> comigrated with the complex obtained with the <protein>purified human p65 NF-kappa B subunit</protein> and binding was inhibited by <protein>I kappa B-alpha</protein> and -beta proteins .


In addition , the 65-kD protein associated with the p50 subunit of NF-kappa B and the kappa B probe to form a complex with the same electrophoretic mobility as the NF-kappa B-DNA complex .

In addition , the <protein>65-kD protein</protein> associated with the <protein>p50 subunit</protein> of <protein>NF-kappa B</protein> and the <dna>kappa B probe</dna> to form a complex with the same electrophoretic mobility as the <protein>NF-kappa B-DNA complex</protein> .


Therefore the rel-related 65-kD protein may represent the p65 subunit of the active NF-kappa B transcription factor complex .

Therefore the <protein>rel-related 65-kD protein</protein> may represent the <protein>p65 subunit</protein> of the <protein>active NF-kappa B transcription factor complex</protein> .


The human myelomonocytic cell line U-937 as a model for studying alterations in steroid-induced monokine gene expression : marked enhancement of lipopolysaccharide-stimulated interleukin-1 beta messenger RNA levels by 1 , 25-dihydroxyvitamin D3 .

The <cell_line>human myelomonocytic cell line U-937</cell_line> as a model for studying alterations in steroid-induced monokine gene expression : marked enhancement of <rna>lipopolysaccharide-stimulated interleukin-1 beta messenger RNA</rna> levels by 1 , 25-dihydroxyvitamin D3 .


The active metabolite of vitamin D , 1 , 25-dihydroxyvitamin D3 [ 1 , 25- ( OH ) 2D3 ] , is a potent regulator of human monocyte/macrophage function in vitro .


To establish a model for 1 , 25- ( OH ) 2D3 regulation of human monocyte monokine synthesis , three human cell lines ( U-937 , THP-1 , and HL-60 ) were examined for : 1 ) the presence of functional 1 , 25- ( OH ) 2D3 receptors ; 2 ) the accumulation of interleukin-1 beta ( IL-1 beta ) mRNA and IL-1 beta protein in response to lipopolysaccharide ( LPS ) ; and 3 ) the regulation of this response by 1 , 25- ( OH ) 2D3 .

To establish a model for 1 , 25- ( OH ) 2D3 regulation of human monocyte monokine synthesis , three <cell_line>human cell lines</cell_line> ( <cell_line>U-937</cell_line> , <cell_line>THP-1</cell_line> , and <cell_line>HL-60</cell_line> ) were examined for : 1 ) the presence of functional 1 , 25- ( OH ) 2D3 receptors ; 2 ) the accumulation of <rna>interleukin-1 beta ( IL-1 beta ) mRNA</rna> and <protein>IL-1 beta protein</protein> in response to lipopolysaccharide ( LPS ) ; and 3 ) the regulation of this response by 1 , 25- ( OH ) 2D3 .


All three cell lines expressed vitamin D receptor and had increased levels of IL-1 beta mRNA in response to LPS .

All three cell lines expressed vitamin D receptor and had increased levels of <rna>IL-1 beta mRNA</rna> in response to LPS .


Preincubation of cells with 1 , 25- ( OH ) 2D3 augmented IL-1 beta mRNA levels only in U-937 and HL-60 cells .

Preincubation of cells with 1 , 25- ( OH ) 2D3 augmented <rna>IL-1 beta mRNA</rna> levels only in <cell_line>U-937</cell_line> and <cell_line>HL-60</cell_line> cells .


From these data , and taking into consideration their state of differentiation and relative ease of culture , U-937 was chosen over HL-60 and THP-1 as the cell line we further characterized .

From these data , and taking into consideration their state of differentiation and relative ease of culture , <cell_line>U-937</cell_line> was chosen over <cell_line>HL-60</cell_line> and <cell_line>THP-1</cell_line> as the cell line we further characterized .


In U-937 cells , optimum time and dose of pretreatment with 1 , 25- ( OH ) 2D3 were determined to be 12-24 h at a receptor saturating concentration of 1 , 25- ( OH ) 2D3 ( 10 nM ) .

In <cell_line>U-937 cells</cell_line> , optimum time and dose of pretreatment with 1 , 25- ( OH ) 2D3 were determined to be 12-24 h at a receptor saturating concentration of 1 , 25- ( OH ) 2D3 ( 10 nM ) .


Preincubation of cells with 1 , 25- ( OH ) 2D3 had no effect on the time course of IL-1 beta mRNA appearance in response to LPS .

Preincubation of cells with 1 , 25- ( OH ) 2D3 had no effect on the time course of <rna>IL-1 beta mRNA</rna> appearance in response to LPS .


However , exposure of U-937 cells to 1 , 25- ( OH ) 2D3 increased by 200 % the level of IL-1 beta mRNA detected and decreased by three orders of magnitude the concentration of LPS required to achieve steady state mRNA levels equivalent to those observed in U-937 cells not preincubated with the hormone.2+o

However , exposure of <cell_line>U-937 cells</cell_line> to 1 , 25- ( OH ) 2D3 increased by 200 % the level of <rna>IL-1 beta mRNA</rna> detected and decreased by three orders of magnitude the concentration of LPS required to achieve steady state mRNA levels equivalent to those observed in <cell_line>U-937 cells</cell_line> not preincubated with the hormone.2+o


Regulation of M-CSF expression by M-CSF : role of protein kinase C and transcription factor NF kappa B .

Regulation of <protein>M-CSF</protein> expression by <protein>M-CSF</protein> : role of <protein>protein kinase C</protein> and <protein>transcription factor NF kappa B</protein> .


Macrophage-colony-stimulating factor ( M-CSF ) , also referred to as CSF-1 , regulates the survival , growth , differentiation and functional activity of monocytes by binding to a single class of high-affinity cell surface receptors , known to be the product of the c-fms protooncogene .

<protein>Macrophage-colony-stimulating factor</protein> ( <protein>M-CSF</protein> ) , also referred to as <protein>CSF-1</protein> , regulates the survival , growth , differentiation and functional activity of <cell_type>monocytes</cell_type> by binding to a single class of <protein>high-affinity cell surface receptors</protein> , known to be the product of the <dna>c-fms protooncogene</dna> .


The detection of both M-CSF and c-fms expression by cells of the monocyte lineage has suggested that M-CSF may act by an autocrine mechanism .

The detection of both M-CSF and c-fms expression by cells of the <cell_type>monocyte lineage</cell_type> has suggested that <protein>M-CSF</protein> may act by an autocrine mechanism .


Interestingly , it has been shown that M-CSF can induce the expression of its own gene .

Interestingly , it has been shown that <protein>M-CSF</protein> can induce the expression of its own gene .


Although sensitivity to M-CSF can be modulated by regulation of receptor expression and function , M-CSF responsiveness is largely determined at a postreceptor level .

Although sensitivity to <protein>M-CSF</protein> can be modulated by regulation of receptor expression and function , <protein>M-CSF</protein> responsiveness is largely determined at a postreceptor level .


To date , little is known about the intracellular pathway of M-CSF signal transduction .

To date , little is known about the intracellular pathway of <protein>M-CSF</protein> signal transduction .


We have therefore investigated the changes in protein kinase C ( PKC ) activity upon exposure of monocytes to M-CSF .

We have therefore investigated the changes in <protein>protein kinase C</protein> ( <protein>PKC</protein> ) activity upon exposure of <cell_type>monocytes</cell_type> to <protein>M-CSF</protein> .


We show that M-CSF activates and translocates PKC .

We show that <protein>M-CSF</protein> activates and translocates <protein>PKC</protein> .


Inhibition of PKC by the isoquinoline derivative H7 abolishes induction of M-CSF by M-CSF .

Inhibition of <protein>PKC</protein> by the isoquinoline derivative H7 abolishes induction of <protein>M-CSF</protein> by <protein>M-CSF</protein> .


Furthermore , activation of PKC was pertussis-toxin-sensitive and was associated with the detection of an NF kappa B protein in nuclear extracts of M-CSF-induced blood monocytes but not in monocytes exposed to medium treatment only .

Furthermore , activation of <protein>PKC</protein> was pertussis-toxin-sensitive and was associated with the detection of an <protein>NF kappa B protein</protein> in nuclear extracts of <cell_line>M-CSF-induced blood monocytes</cell_line> but not in <cell_type>monocytes</cell_type> exposed to medium treatment only .


The results suggest that M-CSF induction of M-CSF involves G proteins , PKC and NF kappa B .

The results suggest that <protein>M-CSF</protein> induction of <protein>M-CSF</protein> involves <protein>G proteins</protein> , <protein>PKC</protein> and <protein>NF kappa B</protein> .


Clone pAT 133 identifies a gene that encodes another human member of a class of growth factor-induced genes with almost identical zinc-finger domains .

<dna>Clone pAT 133</dna> identifies a gene that encodes another human member of a class of <dna>growth factor-induced genes</dna> with almost identical <protein>zinc-finger domains</protein> .


We report the structure and regulation of a gene represented by clone pAT 133 , which is induced upon transition from a resting state ( G0 ) through the early phase of the cell cycle ( G1 ) .

We report the structure and regulation of a gene represented by <dna>clone pAT 133</dna> , which is induced upon transition from a resting state ( G0 ) through the early phase of the cell cycle ( G1 ) .


The pAT 133 gene is immediately induced , with FOS -like kinetics , in human T cells and in fibroblasts .

The <dna>pAT 133 gene</dna> is immediately induced , with <protein>FOS</protein> -like kinetics , in <cell_type>human T cells</cell_type> and in <cell_type>fibroblasts</cell_type> .


Primary structure analysis showed that the encoded protein contains three tandem zinc-finger sequences of the type Cys2-Xaa12-His2 .

Primary structure analysis showed that the encoded protein contains three <protein>tandem zinc-finger sequences</protein> of the type <protein>Cys2-Xaa12-His2</protein> .


This zinc-finger region , which is thought to bind DNA in a sequence-specific manner , is similar ( greater than 80 % on the amino acid level ) to two previously described transcription factors pAT 225/EGR1 and pAT 591/EGR2 .

This <protein>zinc-finger region</protein> , which is thought to bind DNA in a sequence-specific manner , is similar ( greater than 80 % on the amino acid level ) to two previously described <protein>transcription factors</protein> <protein>pAT 225/EGR1</protein> and pAT <protein>591/EGR2</protein> .


Except for the conserved zinc-finger domains , the amino acid sequences of the three proteins are distinct .

Except for the conserved <protein>zinc-finger domains</protein> , the <protein>amino acid sequences</protein> of the three proteins are distinct .


This structural similarity suggests that the pAT 133 gene encodes a transcription factor with a specific biological function .

This structural similarity suggests that the <dna>pAT 133 gene</dna> encodes a <protein>transcription factor</protein> with a specific biological function .


Comparing the regulation of these related zinc-finger-encoding genes showed coordinate induction upon mitogenic stimulation of resting T lymphocytes and of resting fibroblasts .

Comparing the regulation of these related <dna>zinc-finger-encoding genes</dna> showed coordinate induction upon mitogenic stimulation of <cell_type>resting T lymphocytes</cell_type> and of <cell_type>resting fibroblasts</cell_type> .


However , upon transition from a proliferating ( G1 ) to a resting state of the cell cycle the three genes were differently regulated .


In human histiocytic U937 cells mRNA of clone pAT 133 was constitutively expressed , whereas mRNA of pAT 225/EGR1 was induced upon induction of terminal differentiation .

In <rna>human histiocytic U937 cells mRNA</rna> of clone pAT 133 was constitutively expressed , whereas <rna>mRNA</rna> of <protein>pAT 225/EGR1</protein> was induced upon induction of terminal differentiation .


In contrast mRNA representing pAT 591/EGR2 was not expressed in these cells .

In contrast mRNA representing pAT <protein>591/EGR2</protein> was not expressed in these cells .


This difference in gene regulation suggests distinct biological roles in the control of cell proliferation for the respective proteins .


v-erbA overexpression is required to extinguish c-erbA function in erythroid cell differentiation and regulation of the erbA target gene CAII .

<protein>v-erbA</protein> overexpression is required to extinguish <protein>c-erbA</protein> function in <cell_type>erythroid cell</cell_type> differentiation and regulation of the <dna>erbA target gene</dna> <dna>CAII</dna> .


The v-erbA oncoprotein represents a retrovirus-transduced oncogenic version of the thyroid hormone ( T3/T4 ) receptor c-erbA ( type alpha ) .

The <protein>v-erbA oncoprotein</protein> represents a retrovirus-transduced oncogenic version of the <protein>thyroid hormone ( T3/T4 ) receptor</protein> <protein>c-erbA ( type alpha )</protein> .


It contributes to virus-induced erythroleukemia by efficiently arresting differentiation of red cell progenitors and by suppressing transcription of erythrocyte-specific genes .

It contributes to virus-induced erythroleukemia by efficiently arresting differentiation of <cell_type>red cell progenitors</cell_type> and by suppressing transcription of <dna>erythrocyte-specific genes</dna> .


Here , we show that v-erbA and c-erbA bind directly to sequences within the promoter of the erythrocyte-specific carbonic anhydrase II ( CAII ) , a gene whose transcription is efficiently suppressed by v-erbA .

Here , we show that <protein>v-erbA</protein> and <protein>c-erbA</protein> bind directly to sequences within the <dna>promoter</dna> of the <protein>erythrocyte-specific carbonic anhydrase II</protein> ( <dna>CAII</dna> ) , a gene whose transcription is efficiently suppressed by <protein>v-erbA</protein> .


This erbA-binding site confers thyroid hormone responsiveness to a heterologous promoter in transient expression experiments and is a target for efficient down-regulation of CAII transcription by the v-erbA oncoprotein .

This <dna>erbA-binding site</dna> confers thyroid hormone responsiveness to a <dna>heterologous promoter</dna> in transient expression experiments and is a target for efficient down-regulation of <dna>CAII</dna> transcription by the <protein>v-erbA oncoprotein</protein> .


In stably transformed erythroblasts coexpressing the v-erbA oncoprotein and the c-erbA/T3 receptor at an approximately equimolar ratio , c-erbA activity is dominant over v-erbA .

In <cell_line>stably transformed erythroblasts</cell_line> coexpressing the <protein>v-erbA oncoprotein</protein> and the <protein>c-erbA/T3 receptor</protein> at an approximately equimolar ratio , <protein>c-erbA</protein> activity is dominant over <protein>v-erbA</protein> .


T3 efficiently induced erythroid differentiation in these cells , thus overcoming the v-erbA -mediated differentiation arrest .

T3 efficiently induced erythroid differentiation in these cells , thus overcoming the <protein>v-erbA</protein> -mediated differentiation arrest .


Likewise , T3 activated CAII transcription as well as transient expression of a T3-responsive reporter gene containing the CAII-specific erbA-binding site .

Likewise , T3 activated <dna>CAII</dna> transcription as well as transient expression of a <dna>T3-responsive reporter gene</dna> containing the <dna>CAII-specific erbA-binding site</dna> .


The c-erbA -dependent activation of this CAII reporter construct could only be suppressed by very high amounts of v-erbA .

The <protein>c-erbA</protein> -dependent activation of this <dna>CAII reporter construct</dna> could only be suppressed by very high amounts of <protein>v-erbA</protein> .


Our results suggest that overexpression of v-erbA is required for its function as an oncoprotein .

Our results suggest that overexpression of <protein>v-erbA</protein> is required for its function as an <protein>oncoprotein</protein> .


Anti-CD2 receptor antibodies activate the HIV long terminal repeat in T lymphocytes .

<protein>Anti-CD2 receptor antibodies</protein> activate the <dna>HIV long terminal repeat</dna> in <cell_type>T lymphocytes</cell_type> .


The CD2 T lymphocyte glycoprotein surface molecule mediates both cell to cell adhesion and T cell activation , two processes that are involved in the spread of HIV infection .

The <protein>CD2 T lymphocyte glycoprotein surface molecule</protein> mediates both cell to cell adhesion and T cell activation , two processes that are involved in the spread of HIV infection .


Treatment of chronically HIV-infected PBMC with anti-CD2 mAb has been shown to induce the expression of infectious virus from these cultures .

Treatment of <cell_line>chronically HIV-infected PBMC</cell_line> with <protein>anti-CD2 mAb</protein> has been shown to induce the expression of infectious virus from these cultures .


In this study we investigated the mechanisms whereby anti-CD2 antibodies stimulate viral production .

In this study we investigated the mechanisms whereby <protein>anti-CD2 antibodies</protein> stimulate viral production .


We demonstrate that treatment of transiently transfected T lymphocytes with anti-CD2 antibodies results in activation of the HIV long terminal repeat .

We demonstrate that treatment of transiently transfected <cell_type>T lymphocytes</cell_type> with <protein>anti-CD2 antibodies</protein> results in activation of the <dna>HIV long terminal repeat</dna> .


Furthermore , CAT assays using mutated HIV long terminal repeat-CAT constructs and gel shift assays demonstrate that this activation is dependent on the NF-kappa B enhancer .

Furthermore , <protein>CAT</protein> assays using mutated <dna>HIV long terminal repeat-CAT constructs</dna> and gel shift assays demonstrate that this activation is dependent on the <dna>NF-kappa B enhancer</dna> .


These studies suggest that interaction of CD2 with its natural ligand , LFA-3 , may play a role in regulation of HIV expression .

These studies suggest that interaction of <protein>CD2</protein> with its natural ligand , <protein>LFA-3</protein> , may play a role in regulation of HIV expression .


Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF-kappa B transcription factor and HIV-1 .

Reactive oxygen intermediates as apparently widely used messengers in the activation of the <protein>NF-kappa B transcription factor</protein> and HIV-1 .


Hydrogen peroxide and oxygen radicals are agents commonly produced during inflammatory processes .


In this study , we show that micromolar concentrations of H2O2 can induce the expression and replication of HIV-1 in a human T cell line .

In this study , we show that micromolar concentrations of H2O2 can induce the expression and replication of HIV-1 in a <cell_line>human T cell line</cell_line> .


The effect is mediated by the NF-kappa B transcription factor which is potently and rapidly activated by an H2O2 treatment of cells from its inactive cytoplasmic form .

The effect is mediated by the <protein>NF-kappa B</protein> <protein>transcription factor</protein> which is potently and rapidly activated by an H2O2 treatment of cells from its <protein>inactive cytoplasmic form</protein> .


N-acetyl-L-cysteine ( NAC ) , a well characterized antioxidant which counteracts the effects of reactive oxygen intermediates ( ROI ) in living cells , prevented the activation of NF-kappa B by H2O2 .

N-acetyl-L-cysteine ( NAC ) , a well characterized antioxidant which counteracts the effects of reactive oxygen intermediates ( ROI ) in living cells , prevented the activation of <protein>NF-kappa B</protein> by H2O2 .


NAC and other thiol compounds also blocked the activation of NF-kappa B by cycloheximide , double-stranded RNA , calcium ionophore , TNF-alpha , active phorbol ester , interleukin-1 , lipopolysaccharide and lectin .

NAC and other thiol compounds also blocked the activation of <protein>NF-kappa B</protein> by cycloheximide , <rna>double-stranded RNA</rna> , calcium ionophore , <protein>TNF-alpha</protein> , active phorbol ester , <protein>interleukin-1</protein> , lipopolysaccharide and <protein>lectin</protein> .


This suggests that diverse agents thought to activate NF-kappa B by distinct intracellular pathways might all act through a common mechanism involving the synthesis of ROI .

This suggests that diverse agents thought to activate <protein>NF-kappa B</protein> by distinct intracellular pathways might all act through a common mechanism involving the synthesis of ROI .


ROI appear to serve as messengers mediating directly or indirectly the release of the inhibitory subunit I kappa B from NF-kappa B .

ROI appear to serve as messengers mediating directly or indirectly the release of the <protein>inhibitory subunit</protein> <protein>I kappa B</protein> from <protein>NF-kappa B</protein> .


Inhibition of transcription factors belonging to the rel/NF-kappa B family by a transdominant negative mutant .

Inhibition of <protein>transcription factors</protein> belonging to the <protein>rel/NF-kappa B family</protein> by a <protein>transdominant negative mutant</protein> .


The KBF1 factor , which binds to the enhancer A located in the promoter of the mouse MHC class I gene H-2Kb , is indistinguishable from the p50 DNA binding subunit of the transcription factor NF-kappa B , which regulates a series of genes involved in immune and inflammatory responses .

The <protein>KBF1 factor</protein> , which binds to the <dna>enhancer A</dna> located in the <dna>promoter</dna> of the <dna>mouse MHC class I gene</dna> <dna>H-2Kb</dna> , is indistinguishable from the <protein>p50 DNA binding subunit</protein> of the <protein>transcription factor NF-kappa B</protein> , which regulates a series of genes involved in immune and inflammatory responses .


The KBF1/p50 factor binds as a homodimer but can also form heterodimers with the products of other members of the same family , like the c-rel and v-rel ( proto ) oncogenes .

The <protein>KBF1/p50 factor</protein> binds as a <protein>homodimer</protein> but can also form <protein>heterodimers</protein> with the products of other members of the same family , like the <dna>c-rel and v-rel ( proto ) oncogenes</dna> .


The dimerization domain of KBF1/p50 is contained between amino acids 201 and 367 .

The <protein>dimerization domain</protein> of <protein>KBF1/p50</protein> is contained <protein>between amino acids 201 and 367</protein> .


A mutant of KBF1/p50 ( delta SP ) , unable to bind to DNA but able to form homo- or heterodimers , has been constructed .

A mutant of <protein>KBF1/p50</protein> ( <protein>delta SP</protein> ) , unable to bind to DNA but able to form homo- or <protein>heterodimers</protein> , has been constructed .


This protein reduces or abolishes in vitro the DNA binding activity of wild-type proteins of the same family ( KBF1/p50 , c- and v-rel ) .

This protein reduces or abolishes in vitro the DNA binding activity of <protein>wild-type proteins</protein> of the same family ( <protein>KBF1/p50</protein> , <protein>c- and v-rel</protein> ) .


This mutant also functions in vivo as a trans-acting dominant negative regulator : the transcriptional inducibility of the HIV long terminal repeat ( which contains two potential NF-kappa B binding sites ) by phorbol ester ( PMA ) is inhibited when it is co-transfected into CD4+ T cells with the delta SP mutant .

This mutant also functions in vivo as a <protein>trans-acting dominant negative regulator</protein> : the transcriptional inducibility of the <dna>HIV long terminal repeat</dna> ( which contains two potential <protein>NF-kappa B</protein> binding sites ) by phorbol ester ( PMA ) is inhibited when it is co-transfected into <cell_line>CD4+ T cells</cell_line> with the <protein>delta SP</protein> mutant .


Similarly the basal as well as TNF or IL1-induced activity of the MHC class I H-2Kb promoter can be inhibited by this mutant in two different cell lines .

Similarly the basal as well as <protein>TNF</protein> or IL1-induced activity of the <dna>MHC class I H-2Kb promoter</dna> can be inhibited by this mutant in two different <cell_line>cell lines</cell_line> .


These results constitute the first formal demonstration that these genes are regulated by members of the rel/NF-kappa B family .

These results constitute the first formal demonstration that these genes are regulated by members of the <protein>rel/NF-kappa B family</protein> .


Tissue-specific expression of the platelet GPIIb gene .

Tissue-specific expression of the <dna>platelet GPIIb gene</dna> .


One of the major objectives in the study of thrombogenesis is to determine the mechanisms by which a hematopoietic progenitor is activated and committed to the megakaryocytic lineage .

One of the major objectives in the study of thrombogenesis is to determine the mechanisms by which a <cell_type>hematopoietic progenitor</cell_type> is activated and committed to the <cell_type>megakaryocytic lineage</cell_type> .


Recent development of primary cultures of human megakaryocytes and the molecular cloning of genes that are specific to this lineage offer the possibility of getting some insights into the genetic mechanisms that control megakaryocytopoiesis .

Recent development of <cell_line>primary cultures</cell_line> of <cell_line>human megakaryocytes</cell_line> and the molecular cloning of genes that are specific to this lineage offer the possibility of getting some insights into the genetic mechanisms that control megakaryocytopoiesis .


One gene of interest is the glycoprotein IIb ( GPIIb ) gene ; GPIIb , the alpha subunit of the platelet cytoadhesin GPIIb-IIIa , is produced in megakaryocytes at an early stage of the differentiation , whereas the other subunit of this complex , GPIIIa , is expressed in other cells .

One gene of interest is the <dna>glycoprotein IIb ( GPIIb ) gene</dna> ; <dna>GPIIb</dna> , the <protein>alpha subunit</protein> of the <protein>platelet cytoadhesin GPIIb-IIIa</protein> , is produced in <cell_line>megakaryocytes</cell_line> at an early stage of the differentiation , whereas the other subunit of this complex , <protein>GPIIIa</protein> , is expressed in other cells .


For these reasons , the 5'-flanking region of the GPIIb gene was used to identify the regions that interact with DNA-binding nuclear factors .

For these reasons , the <dna>5'-flanking region</dna> of the <dna>GPIIb gene</dna> was used to identify the regions that interact with <protein>DNA-binding nuclear factors</protein> .


A fragment extending from -643 to +33 is capable of controlling the tissue-specific expression of the CAT gene in transfection experiments .

A fragment extending from <dna>-643 to +33</dna> is capable of controlling the tissue-specific expression of the <dna>CAT gene</dna> in transfection experiments .


Within this region , we have identified several sequences that are implicated in DNA protein interactions as shown in DNAse I footprints and gel mobility shift assays .

Within this region , we have identified several sequences that are implicated in DNA protein interactions as shown in <protein>DNAse I</protein> footprints and gel mobility shift assays .


One region , centered at -54 , is similar to a nuclear factor E1-binding site , and a region located at position -233 contains a CCAAT motif .

One region , centered at -54 , is similar to a <dna>nuclear factor E1-binding site</dna> , and a region located at position -233 contains a <dna>CCAAT motif</dna> .


Two domains centered at positions -345 and -540 , respectively , bind proteins that are present in megakaryocytic cells and nonrelated cells as well .

Two domains centered at positions -345 and -540 , respectively , bind proteins that are present in <cell_line>megakaryocytic cells</cell_line> and <cell_type>nonrelated cells</cell_type> as well .


Finally , two other domains , located at positions -460 and -510 , interact with proteins that are only present in megakaryocytic cells .

Finally , two other domains , located at positions -460 and -510 , interact with proteins that are only present in <cell_line>megakaryocytic cells</cell_line> .


In addition , deletion of the region containing these two domains results in a significant decrease of the promoter activity .

In addition , deletion of the region containing these two domains results in a significant decrease of the <dna>promoter</dna> activity .


It is very likely that these domains bind megakaryocyte-specific nuclear proteins acting as positive transcription factors .

It is very likely that these domains bind <protein>megakaryocyte-specific nuclear proteins</protein> acting as <protein>positive transcription factors</protein> .


Lymphocyte glucocorticoid receptor binding during depression and after clinical recovery .

Lymphocyte <protein>glucocorticoid receptor</protein> binding during depression and after clinical recovery .


Lymphocyte glucocorticoid receptor binding parameters were studied in 15 severely depressed patients during depression and after clinical recovery , and in 15 healthy controls .

<protein>Lymphocyte glucocorticoid receptor</protein> binding parameters were studied in 15 severely depressed patients during depression and after clinical recovery , and in 15 healthy controls .


There was no difference in glucocorticoid receptor number or affinity between depressed patients and recovered or control subjects .

There was no difference in <protein>glucocorticoid receptor</protein> number or affinity between depressed patients and recovered or control subjects .


Afternoon ACTH and cortisol concentrations did not differ significantly between the three groups .


No relationship could be established between glucocorticoid receptor binding and antidepressant medication .

No relationship could be established between <protein>glucocorticoid receptor</protein> binding and antidepressant medication .


These data support the view of an impaired ligand-induced plasticity of glucocorticoid receptor regulation rather than the hypothesis of decreased glucocorticoid receptor numbers during depression .

These data support the view of an impaired ligand-induced plasticity of <protein>glucocorticoid receptor</protein> regulation rather than the hypothesis of decreased <protein>glucocorticoid receptor</protein> numbers during depression .


Every enhancer works with every promoter for all the combinations tested : could new regulatory pathways evolve by enhancer shuffling ?

Every enhancer works with every <dna>promoter</dna> for all the combinations tested : could new regulatory pathways evolve by enhancer shuffling ?


The promoters and enhancers of cell type-specific genes are often conserved in evolution , and hence one might expect that a given enhancer has evolved to work best with its own promoter .

The <dna>promoters</dna> and <dna>enhancers</dna> of cell type-specific genes are often conserved in evolution , and hence one might expect that a given <dna>enhancer</dna> has evolved to work best with its own <dna>promoter</dna> .


While this expectation may be realized in some cases , we have not found evidence for it .


A total of 27 combinations of different promoters and enhancers were tested by transfection into cultured cells .

A total of 27 combinations of different <dna>promoters</dna> and <dna>enhancers</dna> were tested by transfection into <cell_line>cultured cells</cell_line> .


We found that the relative efficiency of the enhancers is approximately the same , irrespective of the type of promoter used , i.e. , there was no strong preference for any given enhancer/ promoter combination .

We found that the relative efficiency of the <dna>enhancers</dna> is approximately the same , irrespective of the type of <dna>promoter</dna> used , i.e. , there was no strong preference for any given enhancer/ <dna>promoter</dna> combination .


Notably , we do not see particularly strong transcription when the immunoglobulin kappa enhancer ( or the immunoglobulin heavy chain enhancer ) is used to activate a kappa gene promoter .

Notably , we do not see particularly strong transcription when the <dna>immunoglobulin kappa enhancer</dna> ( or the <dna>immunoglobulin heavy chain enhancer</dna> ) is used to activate a kappa gene <dna>promoter</dna> .


We propose that a generally permissive enhancer/ promoter interaction is of evolutionary benefit for higher eukaryotes : by enhancer shuffling , genes could be easily brought under a new type of inducibility/cell type specificity .

We propose that a generally permissive enhancer/ <dna>promoter</dna> interaction is of evolutionary benefit for higher eukaryotes : by enhancer shuffling , genes could be easily brought under a new type of inducibility/cell type specificity .


Towards a molecular understanding of T-cell differentiation .


Lymphoid differentiation is one of the best studied examples of mammalian development .


Here Hans Clevers and Michael Owen describe how the cloning of the genes that encode T-cell-specific membrane proteins allows the identification of transcription factors that control the expression of these T-cell genes .

Here Hans Clevers and Michael Owen describe how the cloning of the genes that encode <protein>T-cell-specific membrane proteins</protein> allows the identification of <protein>transcription factors</protein> that control the expression of these <dna>T-cell genes</dna> .


Such transcription factors play a key role in the development of the mature T-cell phenotype by functioning as 'master regulators of T-cell differentiation' .

Such <protein>transcription factors</protein> play a key role in the development of the mature T-cell phenotype by functioning as 'master regulators of T-cell differentiation' .


A study on the circadian rhythm of glucocorticoid receptor .

A study on the circadian rhythm of <protein>glucocorticoid receptor</protein> .


Circadian rhythm in glucocorticoid receptor ( GR ) was studied in the rat liver and human peripheral leukocytes .

Circadian rhythm in <protein>glucocorticoid receptor</protein> ( <protein>GR</protein> ) was studied in the <cell_type>rat liver</cell_type> and <cell_type>human peripheral leukocytes</cell_type> .


For rats exposed to a natural environmental photic cycle or a 12L : 12D artificial light regime , peak values of hepatic GR were detected between 23 : 00 and 02 : 00 h .

For rats exposed to a natural environmental photic cycle or a 12L : 12D artificial light regime , peak values of <protein>hepatic GR</protein> were detected between 23 : 00 and 02 : 00 h .


Except for a 4-hour advancement of the peak , a similar circadian rhythm of hepatic GR was detected in rats reared under a reversed lighting regimen ( 12D : 12L ; lights on between 18 : 30 and 06 : 30 h ) .

Except for a 4-hour advancement of the peak , a similar circadian rhythm of <protein>hepatic GR</protein> was detected in rats reared under a reversed lighting regimen ( 12D : 12L ; lights on between 18 : 30 and 06 : 30 h ) .


In human leukocytes , the peak value of GR was found to parallel that of plasma cortisol with high and low values detected at 04 : 00-08 : 00 h and 23 : 00-24 : 00 h , respectively .

In <cell_type>human leukocytes</cell_type> , the peak value of <protein>GR</protein> was found to parallel that of plasma cortisol with high and low values detected at 04 : 00-08 : 00 h and 23 : 00-24 : 00 h , respectively .


In patients suffering from Cushing 's syndrome , the circadian rhythm of plasma cortisol either disappeared or was inverted while that of GR did not significantly deviate from the normal subjects .

In patients suffering from Cushing 's syndrome , the circadian rhythm of plasma cortisol either disappeared or was inverted while that of <protein>GR</protein> did not significantly deviate from the normal subjects .


For apoplexic patients with lesions localized to the base of the brain as indicated by computerized tomography , the diurnal variation of GR was abolished .

For apoplexic patients with lesions localized to the base of the brain as indicated by computerized tomography , the diurnal variation of <protein>GR</protein> was abolished .


Conversely , diurnal rhythmicity persisted in apoplexy patients whose lesions were in the cerebral cortex .


Thus , we postulated that the circadian modification of GR was independent of the diurnal fluctuations in plasma cortisol level or the circadian variations in environmental lighting and that the rhythmicity might be regulated by the 'circadian pacemaker ' located in the human basal brain .

Thus , we postulated that the circadian modification of <protein>GR</protein> was independent of the diurnal fluctuations in plasma cortisol level or the circadian variations in environmental lighting and that the rhythmicity might be regulated by the 'circadian pacemaker ' located in the human basal brain .


These diurnal variations in GR might serve to coordinate the reactivity of the target cells to cortisol because the diurnal rhythms of a GR -mediated response , the fractional inhibition of chemotactic migration rate of polymorphonuclear leukocytes by cortisol , were found to be synchronous with those of GR .

These diurnal variations in <protein>GR</protein> might serve to coordinate the reactivity of the <cell_type>target cells</cell_type> to cortisol because the diurnal rhythms of a <protein>GR</protein> -mediated response , the fractional inhibition of chemotactic migration rate of <cell_type>polymorphonuclear leukocytes</cell_type> by cortisol , were found to be synchronous with those of <protein>GR</protein> .


Transcription factor requirements for U2 snRNA -encoding gene activation in B lymphoid cells .

<protein>Transcription factor</protein> requirements for <rna>U2 snRNA</rna> -encoding gene activation in <cell_type>B lymphoid cells</cell_type> .


Transcription of a human U2 small nuclear RNA ( snRNA ) -encoding gene in HeLa cells requires a distal enhancer element , which is composed of one octamer motif ( Oct ) and three Sp 1-binding sites .

Transcription of a human <dna>U2 small nuclear RNA ( snRNA ) -encoding gene</dna> in <cell_line>HeLa cells</cell_line> requires a <dna>distal enhancer element</dna> , which is composed of one <dna>octamer motif</dna> ( <dna>Oct</dna> ) and three <dna>Sp 1-binding sites</dna> .


To study the transcription factor requirement in B-cells , different U2 enhancer constructions were transfected into the lymphoid cell line , BJA-B .

To study the <protein>transcription factor</protein> requirement in <cell_type>B-cells</cell_type> , different <dna>U2 enhancer constructions</dna> were transfected into the <cell_line>lymphoid cell line</cell_line> , <cell_line>BJA-B</cell_line> .


The results showed that the activation of U2 snRNA transcription in B-cells also requires an enhancer comprising both the Oct and at least one Sp 1-binding site .

The results showed that the activation of U2 snRNA transcription in <cell_type>B-cells</cell_type> also requires an enhancer comprising both the <dna>Oct</dna> and at least one <dna>Sp 1-binding site</dna> .


Deletion of all the Sp 1-binding sites from the enhancer reduces transcription by 80-90 % in HeLa , as well as in BJA-B cells , whereas the removal of the octamer-binding site reduces transcription to levels below detection in both cell types .

Deletion of all the <dna>Sp 1-binding sites</dna> from the <dna>enhancer</dna> reduces transcription by 80-90 % in HeLa , as well as in <cell_line>BJA-B cells</cell_line> , whereas the removal of the <dna>octamer-binding site</dna> reduces transcription to levels below detection in both cell types .


Enhancers containing a single Oct have , nevertheless , the capacity to partially activate U2 snRNA transcription in both HeLa cells , in which only OTF-1 is expressed , and in BJA-B cells in which OTF-2 is the predominantly expressed octamer-binding factor .

Enhancers containing a single <dna>Oct</dna> have , nevertheless , the capacity to partially activate <rna>U2 snRNA</rna> transcription in both <cell_line>HeLa cells</cell_line> , in which only OTF-1 is expressed , and in <cell_line>BJA-B cells</cell_line> in which <protein>OTF-2</protein> is the predominantly expressed <protein>octamer-binding factor</protein> .


The most likely interpretation of our results is that both the ubiquitous transcription factor , OTF-1 , and the B-cell-specific transcription factor , OTF-2 , can activate U2 snRNA transcription .

The most likely interpretation of our results is that both the ubiquitous <protein>transcription factor</protein> , <protein>OTF-1</protein> , and the <protein>B-cell-specific transcription factor</protein> , <protein>OTF-2</protein> , can activate <rna>U2 snRNA</rna> transcription .


The results also revealed a similar functional cooperation between the transcription factors which bind to the Oct and the adjacent Sp 1-binding site in BJA-B cells , as has been observed in HeLa cells , since a template which contains a weak binding site for OTFs expresses wild-type levels of U2 snRNA in both cell types when the weak octamer-binding site is combined with a Sp 1-binding site .

The results also revealed a similar functional cooperation between the transcription factors which bind to the <dna>Oct</dna> and the adjacent <dna>Sp 1-binding site</dna> in <cell_line>BJA-B cells</cell_line> , as has been observed in <cell_line>HeLa cells</cell_line> , since a template which contains a weak binding site for <dna>OTFs</dna> expresses wild-type levels of <rna>U2 snRNA</rna> in both cell types when the weak <dna>octamer-binding site</dna> is combined with a <dna>Sp 1-binding site</dna> .


One base pair change abolishes the T cell-restricted activity of a kB-like proto-enhancer element from the interleukin 2 promoter .

One base pair change abolishes the T cell-restricted activity of a <dna>kB-like proto-enhancer element</dna> from the <dna>interleukin 2 promoter</dna> .


The inducible , T cell-specific enhancers of murine and human Interleukin 2 ( Il-2 ) genes contain the kB-like sequence GGGATTTCACC as an essential cis-acting enhancer motif .

The inducible , <dna>T cell-specific enhancers</dna> of <dna>murine and human Interleukin 2 ( Il-2 ) genes</dna> contain the <dna>kB-like sequence</dna> GGGATTTCACC as an <dna>essential cis-acting enhancer motif</dna> .


When cloned in multiple copies this so-called TCEd ( distal T cell element ) acts as an inducible proto-enhancer element in E14 T lymphoma cells , but not in HeLa cells .

When cloned in multiple copies this so-called <dna>TCEd</dna> ( <dna>distal T cell element</dna> ) acts as an inducible <dna>proto-enhancer element</dna> in <cell_line>E14 T lymphoma cells</cell_line> , but not in <cell_line>HeLa cells</cell_line> .


In extracts of induced , Il-2 secreting El4 cells three individual protein factors bind to TCEd DNA .

In extracts of induced , <cell_line>Il-2 secreting El4 cells</cell_line> three individual <protein>protein factors</protein> bind to <dna>TCEd DNA</dna> .


The binding of the most prominent factor , named TCF-1 ( T cell factor 1 ) , is correlated with the proto-enhancer activity of TCEd .

The binding of the most prominent factor , named <protein>TCF-1</protein> ( <protein>T cell factor 1</protein> ) , is correlated with the proto-enhancer activity of TCEd .


TCF-1 consists of two polypeptides of about 50 kD and 105 kD ; the former seems to be related to the 50 kD polypeptide of NF-kB .

<protein>TCF-1</protein> consists of two polypeptides of about 50 kD and 105 kD ; the former seems to be related to the <protein>50 kD polypeptide</protein> of <protein>NF-kB</protein> .


Purified NF-kB is also able to bind to the TCEd , but TCF-1 binds stronger than NF-kB to TCEd DNA .

<protein>Purified NF-kB</protein> is also able to bind to the <dna>TCEd</dna> , but <protein>TCF-1</protein> binds stronger than <protein>NF-kB</protein> to <dna>TCEd</dna> DNA .


The conversion of the TCEd to a 'perfect ' NF-kB binding site leads to a tighter binding of NF-kB to TCEd DNA and , as a functional consequence , to the activity of the 'converted ' TCEd motifs in HeLa cells .

The conversion of the <dna>TCEd</dna> to a <dna>'perfect ' NF-kB binding site</dna> leads to a tighter binding of <protein>NF-kB</protein> to <dna>TCEd DNA</dna> and , as a functional consequence , to the activity of the <dna>'converted ' TCEd motifs</dna> in <cell_line>HeLa cells</cell_line> .


Thus , the substitution of the underlined A residue to a C within the GGGATTTCACC motif abolishes its T cell-restricted activity and leads to its functioning in both El4 cells and HeLa cells .

Thus , the substitution of the underlined A residue to a C within the <dna>GGGATTTCACC motif</dna> abolishes its T cell-restricted activity and leads to its functioning in both <cell_line>El4 cells</cell_line> and <cell_line>HeLa cells</cell_line> .


These results indicate that lymphocyte-specific factors binding to the TCEd are involved in the control of T cell specific-transcription of the Il-2 gene .

These results indicate that lymphocyte-specific factors binding to the <dna>TCEd</dna> are involved in the control of T cell specific-transcription of the <dna>Il-2 gene</dna> .


An erythroid specific enhancer upstream to the gene encoding the cell-type specific transcription factor GATA-1 .

An <dna>erythroid specific enhancer</dna> upstream to the gene encoding the <protein>cell-type specific transcription factor</protein> <protein>GATA-1</protein> .


The transcription factor GATA-1 is expressed in a subset of hemopoietic cells , where it mediates the cell-type specific expression of several genes .

The <protein>transcription factor</protein> <protein>GATA-1</protein> is expressed in a subset of <cell_type>hemopoietic cells</cell_type> , where it mediates the cell-type specific expression of several genes .


We have cloned the mouse and human GATA-1 genes .

We have cloned the mouse and human <dna>GATA-1 genes</dna> .


A region upstream to the first exon , and highly conserved between mouse and man , acts as an erythroid specific enhancer in transient assays , if linked to the GATA-1 or to the SV40 promoter .

A region upstream to the first exon , and highly conserved between mouse and man , acts as an erythroid specific enhancer in transient assays , if linked to the <protein>GATA-1</protein> or to the SV40 promoter .


The activity of the enhancer is almost completely dependent on the integrity of a dimeric GATA-1 binding site .

The activity of the enhancer is almost completely dependent on the integrity of a <dna>dimeric GATA-1 binding site</dna> .


Demonstration of a 1 , 25-dihydroxyvitamin D3-responsive protein in human lymphocytes : immunologic crossreactivity and inverse regulation with the vitamin D receptor .

Demonstration of a <protein>1 , 25-dihydroxyvitamin D3-responsive protein</protein> in <cell_type>human lymphocytes</cell_type> : immunologic crossreactivity and inverse regulation with the <protein>vitamin D receptor</protein> .


Using Western blot analysis with a monoclonal antibody recognizing a 17-amino acid epitope of the 1 , 25-dihydroxyvitamin D3 [ 1 , 25 ( OH ) 2D3 ] receptor , we have detected two crossreacting proteins in activated normal human lymphocytes .

Using Western blot analysis with a <protein>monoclonal antibody</protein> recognizing a <protein>17-amino acid epitope</protein> of the <protein>1 , 25-dihydroxyvitamin D3 [ 1 , 25 ( OH ) 2D3 ] receptor</protein> , we have detected two <protein>crossreacting proteins</protein> in <cell_type>activated normal human lymphocytes</cell_type> .


The smaller of the two proteins ( 50 kDa ) was indistinguishable from the classical 1 , 25 ( OH ) 2D3 receptor and , similar to the classical 1 , 25 ( OH ) 2D3 receptor , was upregulated in a dose-dependent fashion by 1 , 25 ( OH ) 2D3 .

The smaller of the two proteins ( <protein>50 kDa</protein> ) was indistinguishable from the classical <protein>1 , 25 ( OH ) 2D3 receptor</protein> and , similar to the classical <protein>1 , 25 ( OH ) 2D3 receptor</protein> , was upregulated in a dose-dependent fashion by 1 , 25 ( OH ) 2D3 .


The larger crossreacting protein exhibited an electrophoretic mobility of 80 kDa , was localized in the cell cytosol , and appeared to be specific for activated lymphocytes since it was not detected in several other human cells including monocytes .

The larger crossreacting protein exhibited an electrophoretic mobility of 80 kDa , was localized in the cell cytosol , and appeared to be specific for <cell_type>activated lymphocytes</cell_type> since it was not detected in several other <cell_type>human cells</cell_type> including <cell_type>monocytes</cell_type> .


More strikingly , the 80-kDa protein was downregulated in a dose-dependent fashion by 1 , 25 ( OH ) 2D3 ; this effect was independent of the mode of lymphocyte activation and specific for the 1 , 25 ( OH ) 2D3 metabolite of vitamin D3 .

More strikingly , the <protein>80-kDa protein</protein> was downregulated in a dose-dependent fashion by 1 , 25 ( OH ) 2D3 ; this effect was independent of the mode of lymphocyte activation and specific for the 1 , 25 ( OH ) 2D3 metabolite of vitamin D3 .


However , two potent immunosuppressive agents , glucocorticoids and cyclosporin A , also inhibited the 80-kDa protein .

However , two potent immunosuppressive agents , glucocorticoids and cyclosporin A , also inhibited the <protein>80-kDa protein</protein> .


Regulation of glucocorticoid receptors in human mononuclear cells : effects of glucocorticoid treatment , Cushing 's disease and ketoconazole .

Regulation of <protein>glucocorticoid receptors</protein> in <cell_type>human mononuclear cells</cell_type> : effects of glucocorticoid treatment , Cushing 's disease and ketoconazole .


Glucocorticoid receptors ( GcR ) were determined by a whole cell assay in human mononulear leukocytes ( hMNL ) from control subjects , patients receiving glucocorticoid therapy for systemic diseases and Cushing 's disease patients with or without ketoconazole therapy .

<protein>Glucocorticoid receptors</protein> ( <protein>GcR</protein> ) were determined by a whole cell assay in <cell_type>human mononulear leukocytes</cell_type> ( <cell_type>hMNL</cell_type> ) from control subjects , patients receiving glucocorticoid therapy for systemic diseases and Cushing 's disease patients with or without ketoconazole therapy .


Prolonged corticosteroid treatment resulted in down-regulation of GcR , while the mean level of GcR in Cushing 's disease was normal .

Prolonged corticosteroid treatment resulted in down-regulation of <protein>GcR</protein> , while the mean level of <protein>GcR</protein> in Cushing 's disease was normal .


In this group , however , receptor levels and morning plasma cortisol values showed a negative correlation , indicating a subtle down-regulatory effect .


Furthermore , GcR were unaltered after these patients received ketoconazole , in spite of a marked reduction in morning plasma cortisol and urinary free cortisol .

Furthermore , <protein>GcR</protein> were unaltered after these patients received ketoconazole , in spite of a marked reduction in morning plasma cortisol and urinary free cortisol .


We also observed that ketoconazole was a weak competitor of GcR in intact cells , although it significantly inhibited [ 3H ] dexamethasone binding in cytosolic preparations from rat tissues .

We also observed that ketoconazole was a weak competitor of <protein>GcR</protein> in <cell_type>intact cells</cell_type> , although it significantly inhibited [ 3H ] dexamethasone binding in cytosolic preparations from rat tissues .


The results suggested that GcR in hMNL are down-regulated by synthetic steroids given in vivo , but they showed very mild down-regulation in hypercortisolemic patients suffering from Cushing 's disease .

The results suggested that <protein>GcR</protein> in <cell_type>hMNL</cell_type> are down-regulated by synthetic steroids given in vivo , but they showed very mild down-regulation in hypercortisolemic patients suffering from Cushing 's disease .


Finally , we did not observed either up-regulation or antagonism of GcR by ketoconazole treatment , at the time that cortisol levels of patients with Cushing 's disease were reduced .

Finally , we did not observed either up-regulation or antagonism of <protein>GcR</protein> by ketoconazole treatment , at the time that cortisol levels of patients with Cushing 's disease were reduced .


This indicates that the beneficial effects of ketoconazole in Cushing 's disease are due to adrenal cortisol suppression and not to interaction with GcR of target cells , and that the process of GcR regulation in hMNL is a complex phenomenon awaiting further elucidation .

This indicates that the beneficial effects of ketoconazole in Cushing 's disease are due to adrenal cortisol suppression and not to interaction with <protein>GcR</protein> of <cell_type>target cells</cell_type> , and that the process of <protein>GcR</protein> regulation in <cell_type>hMNL</cell_type> is a complex phenomenon awaiting further elucidation .


HTLV-1 Tax induces expression of various immediate early serum responsive genes .

<protein>HTLV-1 Tax</protein> induces expression of various <dna>immediate early serum responsive genes</dna> .


Human T-cell leukemia virus type 1 ( HTLV-1 ) is an etiological agent of adult T-cell leukemia ( ATL ) .


We showed here by mobility-shift assay that T-cell lines transformed with the virus contained high levels of AP-1 activities .

We showed here by mobility-shift assay that <cell_line>T-cell lines</cell_line> transformed with the virus contained high levels of <protein>AP-1</protein> activities .


Consistent with this result , these cell lines expressed increased levels of mRNAs encoding the AP-1 proteins , c-Fos , Fra-1 , c-Jun , JunB , and JunD .

Consistent with this result , these cell lines expressed increased levels of <rna>mRNAs</rna> encoding the <protein>AP-1 proteins</protein> , <protein>c-Fos</protein> , <protein>Fra-1</protein> , <protein>c-Jun</protein> , <protein>JunB</protein> , and <protein>JunD</protein> .


Previously , transcription of the c-fos gene has been reported to be transactivated by the viral transcription factor , Tax1 .

Previously , transcription of the <dna>c-fos gene</dna> has been reported to be transactivated by the <protein>viral transcription factor</protein> , <protein>Tax1</protein> .


By using the human T-cell line ( JPX-9 ) , in which expression of the Tax1 is inducible , we showed that expression of mRNAs for Fra-1 , c-Jun , and JunD was also transactivated by Tax1 .

By using the <cell_line>human T-cell line</cell_line> ( <cell_line>JPX-9</cell_line> ) , in which expression of the <protein>Tax1</protein> is inducible , we showed that expression of <rna>mRNAs</rna> for <protein>Fra-1</protein> , <protein>c-Jun</protein> , and <protein>JunD</protein> was also transactivated by <protein>Tax1</protein> .


Moreover , Tax1 activated expression of two other transcription factors having zinc finger motifs , Egr-1 and Egr-2 , in the same cells .

Moreover , <protein>Tax1</protein> activated expression of two other <protein>transcription factors</protein> having <protein>zinc finger motifs</protein> , <protein>Egr-1</protein> and <protein>Egr-2</protein> , in the same cells .


The Tax1-inducible transcription factors identified here are encoded by the members of immediate early genes under the control of growth signals .

The <protein>Tax1-inducible transcription factors</protein> identified here are encoded by the members of <dna>immediate early genes</dna> under the control of growth signals .


Thus , Tax1 was suggested to replace growth signals , at least in part , by this mechanism .

Thus , <protein>Tax1</protein> was suggested to replace growth signals , at least in part , by this mechanism .


Glucocorticoid receptor characteristics in monocytes of patients with corticosteroid-resistant bronchial asthma .

<protein>Glucocorticoid receptor</protein> characteristics in <cell_type>monocytes</cell_type> of patients with corticosteroid-resistant bronchial asthma .


The mechanism of corticosteroid resistance in bronchial asthma has been studied by determining the rank order of potency for different corticosteroids in inhibiting the generation of a 3 kD molecule from peripheral blood monocytes isolated from corticosteroid-sensitive ( CS ) and corticosteroid-resistant ( CR ) asthmatic subjects , which augments leukotriene B4 ( LTB4 ) generation by human neutrophils ( PMN ) stimulated by calcium ionophore .

The mechanism of corticosteroid resistance in bronchial asthma has been studied by determining the rank order of potency for different corticosteroids in inhibiting the generation of a 3 kD molecule from <cell_type>peripheral blood monocytes</cell_type> isolated from corticosteroid-sensitive ( CS ) and corticosteroid-resistant ( CR ) asthmatic subjects , which augments leukotriene B4 ( <protein>LTB4</protein> ) generation by <cell_type>human neutrophils</cell_type> ( <cell_type>PMN</cell_type> ) stimulated by calcium ionophore .


In addition , binding studies with ( 3H ) dexamethasone have been performed to determine the dissociation constant ( Kd ) and receptor numbers ( Ro ) in the monocytes of these two groups of subjects .

In addition , binding studies with ( 3H ) dexamethasone have been performed to determine the dissociation constant ( Kd ) and receptor numbers ( Ro ) in the <cell_type>monocytes</cell_type> of these two groups of subjects .


The concentration of corticosteroid producing 50 % inhibition ( IC50 ) was 600 nM , 70 nM , and 0.5 nM for hydrocortisone , methylprednisolone , and dexamethasone , respectively , in monocytes from CS individuals .

The concentration of corticosteroid producing 50 % inhibition ( IC50 ) was 600 nM , 70 nM , and 0.5 nM for hydrocortisone , methylprednisolone , and dexamethasone , respectively , in <cell_type>monocytes</cell_type> from CS individuals .


There was only weak inhibition of the generation of enhancing activity by the corticosteroids in the CR asthmatic individuals .


The dexamethasone Kd was 2.45 +/- 0.58 nM ( mean +/- SEM , n = 6 ) in the CS group and 1.6 +/- 0.35 nM ( mean +/- SEM , n = 6 ) in the CR group of patients ( p = 0.14 ) .


The Ro in the CS group was 3 , 605 +/- 984 binding sites per nucleus ( mean +/- SEM , n = 6 ) and 4 , 757 +/- 692 binding sites per nucleus ( mean +/- SEM , n = 6 ) in the CR group ( p = 0.23 ) .


These findings indicate that corticosteroid resistance in bronchial asthma can not be explained by abnormalities in corticosteroid receptor characteristics .


Vitamin D receptor expression in human lymphocytes .

<protein>Vitamin D receptor</protein> expression in <cell_type>human lymphocytes</cell_type> .


Signal requirements and characterization by western blots and DNA sequencing .


The signals controlling the expression of the receptor protein for 1 alpha , 25-dihydroxyvitamin D3 in normal human lymphocytes and the relationship of this protein to the classical vitamin D receptor were examined .

The signals controlling the expression of the <protein>receptor protein</protein> for 1 alpha , 25-dihydroxyvitamin D3 in normal <cell_type>human lymphocytes</cell_type> and the relationship of this protein to the <protein>classical vitamin D receptor</protein> were examined .


Lymphocytes activated with the OKT3 antibody to the T-cell antigen receptor expressed fewer binding sites as compared to lymphocytes that were activated by the polyclonal activator phytohemagglutinin ( PHA ) .

<cell_type>Lymphocytes</cell_type> activated with the <protein>OKT3 antibody</protein> to the <protein>T-cell antigen receptor</protein> expressed fewer binding sites as compared to <cell_type>lymphocytes</cell_type> that were activated by the polyclonal activator <protein>phytohemagglutinin</protein> ( <protein>PHA</protein> ) .


However , combination of OKT3 and phorbol myristate acetate produced a concentration of binding sites similar to the PHA-activated cells .

However , combination of <protein>OKT3</protein> and phorbol myristate acetate produced a concentration of binding sites similar to the <cell_line>PHA-activated cells</cell_line> .


The receptor from OKT3 and OKT3 + phorbol myristate acetate -activated lymphocytes exhibited decreased binding to DNA-cellulose compared to PHA-activated lymphocytes .

The receptor from <protein>OKT3</protein> and <protein>OKT3</protein> + phorbol myristate acetate <cell_type>-activated lymphocytes</cell_type> exhibited decreased binding to DNA-cellulose compared to <cell_type>PHA-activated lymphocytes</cell_type> .


In lymphocytes activated either by PHA or OKT3 ( but not in resting cells ) , a 50-kDa species cross-reacting with a monoclonal antibody against the intestinal vitamin D receptor was detected .

In <cell_type>lymphocytes</cell_type> activated either by <protein>PHA</protein> or <protein>OKT3</protein> ( but not in <cell_type>resting cells</cell_type> ) , a <protein>50-kDa species</protein> cross-reacting with a <protein>monoclonal antibody</protein> against the <protein>intestinal vitamin D receptor</protein> was detected .


Finally , RNA from activated lymphocytes was amplified by polymerase chain reaction using oligonucleotide primers flanking the 196 base pair long region encoding the DNA-binding domain of the human intestinal receptor .

Finally , RNA from <cell_type>activated lymphocytes</cell_type> was amplified by polymerase chain reaction using oligonucleotide primers flanking the <dna>196 base pair long region</dna> encoding the <protein>DNA-binding domain</protein> of the <protein>human intestinal receptor</protein> .


The amplified product showed an identical nucleotide sequence to the DNA-binding domain of the human intestinal receptor .

The amplified product showed an <dna>identical nucleotide sequence</dna> to the <protein>DNA-binding domain</protein> of the <protein>human intestinal receptor</protein> .


These findings suggest that expression of the 1 , 25- ( OH ) 2D3 receptor in lymphocytes is triggered by distinct and contingent signals , and that the protein and the mRNA encoding it are identical to the classical vitamin D receptor .

These findings suggest that expression of the <protein>1 , 25- ( OH ) 2D3 receptor</protein> in <cell_type>lymphocytes</cell_type> is triggered by distinct and contingent signals , and that the protein and the <rna>mRNA</rna> encoding it are identical to the <protein>classical vitamin D receptor</protein> .


Comparison of constitutive and inducible transcriptional enhancement mediated by kappa B-related sequences : modulation of activity in B cells by human T-cell leukemia virus type I tax gene .

Comparison of constitutive and inducible transcriptional enhancement mediated by <dna>kappa B-related sequences</dna> : modulation of activity in <cell_type>B cells</cell_type> by <dna>human T-cell leukemia virus type I tax gene</dna> .


The kappa B sequence ( GGGACTTTCC ) binds a factor , NF-kappa B , that is constitutively found in its functional , DNA binding form only in B lymphocytes .

The <dna>kappa B sequence</dna> ( GGGACTTTCC ) binds a factor , <protein>NF-kappa B</protein> , that is constitutively found in its functional , DNA binding form only in <cell_type>B lymphocytes</cell_type> .


A factor with apparently indistinguishable sequence specificity can be induced in many other cell types , where it is used to regulate inducible gene expression .


For example , kappa B-related sequences have been shown to be important for the transcription of a few inducible genes , such as the interleukin 2 receptor alpha-chain gene and the beta-interferon gene .

For example , <dna>kappa B-related sequences</dna> have been shown to be important for the transcription of a few inducible genes , such as the <dna>interleukin 2 receptor alpha-chain gene</dna> and the <dna>beta-interferon gene</dna> .


However , these genes are not constitutively active in B lymphocytes , suggesting that other regulatory mechanisms must play a role in determining the patterns of expression .

However , these genes are not constitutively active in <cell_type>B lymphocytes</cell_type> , suggesting that other regulatory mechanisms must play a role in determining the patterns of expression .


We have investigated the constitutive and inducible transcriptional activity mediated by five kappa B-related sequence elements in two different cell types .

We have investigated the constitutive and inducible transcriptional activity mediated by five <dna>kappa B-related sequence elements</dna> in two different cell types .


We show that in S194 plasma cells the activity of each element correlates well with the relative affinity of B-cell-derived NF-kappa B for that element .

We show that in <cell_line>S194 plasma cells</cell_line> the activity of each element correlates well with the relative affinity of <protein>B-cell-derived NF-kappa B</protein> for that element .


This leads to significantly lower transcription enhancement by sites derived from the interleukin 2 receptor or T-cell receptor genes in S194 cells .

This leads to significantly lower transcription enhancement by sites derived from the <protein>interleukin 2 receptor</protein> or <dna>T-cell receptor genes</dna> in <cell_line>S194 cells</cell_line> .