Inhibition of human immunodeficiency virus type 1 replication by a Tat-activated , transduced interferon gene : targeted expression to human immunodeficiency virus type 1-infected cells .

Inhibition of human immunodeficiency virus type 1 replication by a <dna>Tat-activated , transduced interferon gene</dna> : targeted expression to <cell_type>human immunodeficiency virus type 1-infected cells</cell_type> .


We have examined the feasibility of using interferon ( IFN ) gene transfer as a novel approach to anti-human immunodeficiency virus type 1 ( HIV-1 ) therapy in this study .

We have examined the feasibility of using <protein>interferon</protein> ( <protein>IFN</protein> ) gene transfer as a novel approach to anti-human immunodeficiency virus type 1 ( HIV-1 ) therapy in this study .


To limit expression of a transduced HIV-1 long terminal repeat ( LTR ) -IFNA2 ( the new approved nomenclature for IFN genes is used throughout this article ) hybrid gene to the HIV-1-infected cells , HIV-1 LTR was modified .

To limit expression of a transduced <dna>HIV-1 long terminal repeat ( LTR ) -IFNA2</dna> ( the new approved nomenclature for <dna>IFN genes</dna> is used throughout this article ) <dna>hybrid gene</dna> to the <cell_line>HIV-1-infected cells</cell_line> , <dna>HIV-1 LTR</dna> was modified .


Deletion of the NF-kappa B elements of the HIV-1 LTR significantly inhibited Tat -mediated transactivation in T-cell lines , as well as in a monocyte line , U937 .

Deletion of the <dna>NF-kappa B elements</dna> of the <dna>HIV-1 LTR</dna> significantly inhibited <protein>Tat</protein> -mediated transactivation in <cell_line>T-cell lines</cell_line> , as well as in a <cell_line>monocyte line</cell_line> , <cell_line>U937</cell_line> .


Replacement of the NF-kappa B elements in the HIV-1 LTR by a DNA fragment derived from the 5'-flanking region of IFN-stimulated gene 15 ( ISG15 ) , containing the IFN-stimulated response element , partially restored Tat -mediated activation of LTR in T cells as well as in monocytes .

Replacement of the <dna>NF-kappa B elements</dna> in the <dna>HIV-1 LTR</dna> by a <dna>DNA fragment</dna> derived from the <dna>5'-flanking region</dna> of <dna>IFN-stimulated gene 15</dna> ( <dna>ISG15</dna> ) , containing the <dna>IFN-stimulated response element</dna> , partially restored <protein>Tat</protein> -mediated activation of <dna>LTR</dna> in <cell_type>T cells</cell_type> as well as in <cell_type>monocytes</cell_type> .


Insertion of this chimeric promoter ( ISG15 LTR ) upstream of the human IFNA2 gene directed high levels of IFN synthesis in Tat-expressing cells , while this promoter was not responsive to tumor necrosis factor alpha -mediated activation .

Insertion of this <dna>chimeric promoter</dna> ( <dna>ISG15 LTR</dna> ) upstream of the <dna>human IFNA2 gene</dna> directed high levels of <protein>IFN</protein> synthesis in <cell_line>Tat-expressing cells</cell_line> , while this promoter was not responsive to <protein>tumor necrosis factor alpha</protein> -mediated activation .


ISG15-LTR-IFN hybrid gene inserted into the retrovirus vector was transduced into Jurkat and U937 cells .

<dna>ISG15-LTR-IFN hybrid gene</dna> inserted into the <dna>retrovirus vector</dna> was transduced into <cell_line>Jurkat</cell_line> and <cell_line>U937 cells</cell_line> .


Selected transfected clones produced low levels of IFN A ( IFNA ) constitutively , and their abilities to express interleukin-2 and interleukin-2 receptor upon stimulation with phytohemagglutinin and phorbol myristate acetate were retained .

Selected <cell_line>transfected clones</cell_line> produced low levels of <protein>IFN A</protein> ( <protein>IFNA</protein> ) constitutively , and their abilities to express <protein>interleukin-2</protein> and <protein>interleukin-2 receptor</protein> upon stimulation with <protein>phytohemagglutinin</protein> and phorbol myristate acetate were retained .


Enhancement of IFNA synthesis observed upon HIV-1 infection resulted in significant inhibition of HIV-1 replication for a period of at least 30 days .

Enhancement of <protein>IFNA</protein> synthesis observed upon HIV-1 infection resulted in significant inhibition of HIV-1 replication for a period of at least 30 days .


Virus isolated from IFNA-producing cells was able to replicate in the U937 cells but did not replicate efficiently in U937 cells transduced with the IFNA gene .

Virus isolated from <cell_line>IFNA-producing cells</cell_line> was able to replicate in the <cell_line>U937 cells</cell_line> but did not replicate efficiently in <cell_line>U937 cells</cell_line> transduced with the <dna>IFNA gene</dna> .


These results suggest that targeting IFN synthesis to HIV-1-infected cells is an attainable goal and that autocrine IFN synthesis results in a long-lasting and permanent suppression of HIV-1 replication .

These results suggest that targeting <protein>IFN</protein> synthesis to <cell_line>HIV-1-infected cells</cell_line> is an attainable goal and that autocrine <protein>IFN</protein> synthesis results in a long-lasting and permanent suppression of HIV-1 replication .


Chromosomal localization of two KOX zinc finger genes on chromosome bands 7q21-q22 .

Chromosomal localization of two <dna>KOX zinc finger genes</dna> on <dna>chromosome bands</dna> <dna>7q21-q22</dna> .


Human cDNAs encoding Kruppel-type zinc finger domains , designated KOX 1-32 , have been cloned from human T lymphocyte cell line libraries .

<dna>Human cDNAs</dna> encoding <protein>Kruppel-type zinc finger domains</protein> , designated <dna>KOX 1-32</dna> , have been cloned from <dna>human T lymphocyte cell line libraries</dna> .


We report here the regional localizations by in situ hybridization of KOX 18 and KOX 25 on chromosome bands 7q21q22 .

We report here the regional localizations by in situ hybridization of <dna>KOX 18</dna> and <dna>KOX 25</dna> on <dna>chromosome bands 7q21q22</dna> .


Pulse-field gel electrophoresis ( PFGE ) analysis showed that these genes are physically located within a DNA fragment of 250 kb .

Pulse-field gel electrophoresis ( PFGE ) analysis showed that these genes are physically located within a <dna>DNA fragment</dna> of <dna>250 kb</dna> .


The genes KOX 4 and KOX 9 , mapped on chromosome 8q24 , were found to be located within a DNA fragment of 450 kb .

The genes <dna>KOX 4</dna> and <dna>KOX 9</dna> , mapped on <dna>chromosome 8q24</dna> , were found to be located within a <dna>DNA fragment</dna> of 450 kb .


From the present and previous data , eighteen different KOX genes have been located at least two by two within nine DNA fragments of 200 to 580 kb .

From the present and previous data , eighteen different <dna>KOX genes</dna> have been located at least two by two within nine <dna>DNA fragments</dna> of 200 to 580 kb .


Influence of age on the production of Fos and Jun by influenza virus-exposed T cells .

Influence of age on the production of <protein>Fos</protein> and <protein>Jun</protein> by <cell_type>influenza virus-exposed T cells</cell_type> .


This study investigated age-related T cell responses after in vitro exposure to influenza A virus .


Mononuclear leukocytes from young or elderly persons were sham-exposed or exposed to influenza virus for 1 , 24 , and 72 h .

<cell_type>Mononuclear leukocytes</cell_type> from young or elderly persons were sham-exposed or exposed to influenza virus for 1 , 24 , and 72 h .


Immunofluorescent staining and flow cytometric analysis were then used to detect T cells producing the transcriptional regulating proteins Fos and Jun .

Immunofluorescent staining and flow cytometric analysis were then used to detect <cell_type>T cells</cell_type> producing the <protein>transcriptional regulating proteins</protein> <protein>Fos</protein> and <protein>Jun</protein> .


Fewer virus-exposed cells from elderly donors stained for Fos and Jun at each data point compared with cells from young donors .

Fewer virus-exposed cells from elderly donors stained for <protein>Fos</protein> and <protein>Jun</protein> at each data point compared with cells from young donors .


Flow cytometric analysis also showed that at 72 h of virus exposure fewer T cells from the elderly produced interferon-gamma ( IFN-gamma ) , suggesting a link between the magnitude of the Fos and Jun and IFN-gamma responses .

Flow cytometric analysis also showed that at 72 h of virus exposure fewer <cell_type>T cells</cell_type> from the elderly produced <protein>interferon-gamma</protein> ( <protein>IFN-gamma</protein> ) , suggesting a link between the magnitude of the <protein>Fos</protein> and <protein>Jun</protein> and <protein>IFN-gamma</protein> responses .


Thus , failure of virus-exposed T cells to produce Fos and Jun could contribute to the increase in illness due to influenza virus in the elderly .

Thus , failure of <cell_type>virus-exposed T cells</cell_type> to produce <protein>Fos</protein> and <protein>Jun</protein> could contribute to the increase in illness due to influenza virus in the elderly .


The regulation of HIV by retinoic acid correlates with cellular expression of the retinoic acid receptors .

The regulation of HIV by retinoic acid correlates with cellular expression of the <protein>retinoic acid receptors</protein> .


OBJECTIVES : To analyze the effect of retinoic acids ( RA ) on HIV-1 expression and correlate this effect with expression levels of RA receptors ( RARs ) in T-lymphoid and monocytoid cell lines .

OBJECTIVES : To analyze the effect of retinoic acids ( RA ) on HIV-1 expression and correlate this effect with expression levels of <protein>RA receptors</protein> ( <protein>RARs</protein> ) in <cell_line>T-lymphoid and monocytoid cell lines</cell_line> .


DESIGN AND METHODS : The effect of all-trans and 9-cis RA on HIV-1 production in T-lymphoid ( H9 , CEM ) and monocytoid ( U937 , THP-1 ) cell lines was measured during acute and chronic infection .

DESIGN AND METHODS : The effect of all-trans and 9-cis RA on HIV-1 production in <cell_line>T-lymphoid</cell_line> ( <cell_line>H9 , CEM</cell_line> ) and <cell_line>monocytoid ( U937 , THP-1 ) cell lines</cell_line> was measured during acute and chronic infection .


The expression levels of human RAR alpha ( hRAR alpha , receptor for all-trans RA ) and the human retinoid-X receptor alpha ( hRXR alpha receptor for 9-cis RA ) were determined by Northern blot analysis .

The expression levels of <protein>human RAR alpha</protein> ( <protein>hRAR alpha , receptor</protein> for all-trans RA ) and the <protein>human retinoid-X receptor alpha</protein> ( <protein>hRXR alpha receptor</protein> for 9-cis RA ) were determined by Northern blot analysis .


RESULTS : Both all-trans and 9-cis RA inhibited virus replication in HIV-1 IIIB-infected monocytoid cells , in the presence and absence of the co-stimulatory agent phorbol myristate acetate ( PMA ) .

RESULTS : Both all-trans and 9-cis RA inhibited virus replication in <cell_line>HIV-1 IIIB-infected monocytoid cells</cell_line> , in the presence and absence of the co-stimulatory agent phorbol myristate acetate ( PMA ) .


The retinoids had weak or no stimulatory effects on HIV production by T-cell lines .

The retinoids had weak or no stimulatory effects on HIV production by <cell_line>T-cell lines</cell_line> .


HIV production by PMA-stimulated T-cell lines was inhibited by these retinoids .

HIV production by <cell_line>PMA-stimulated T-cell lines</cell_line> was inhibited by these retinoids .


The 9-cis RA was generally more effective than all-trans RA in inhibiting HIV production and in combination generally more effective than the single agents alone .


Human RAR alpha was expressed in H9 , U937 and THP-1 cells , but almost undetectable in CEM cells .

<protein>Human RAR alpha</protein> was expressed in <cell_line>H9</cell_line> , <cell_line>U937</cell_line> and <cell_line>THP-1 cells</cell_line> , but almost undetectable in <cell_line>CEM cells</cell_line> .


Human RXR alpha was significantly expressed in U937 and THP-1 cells , weakly expressed in H9 cells and not detectable in CEM cells .

<protein>Human RXR alpha</protein> was significantly expressed in <cell_line>U937 and THP-1 cells</cell_line> , weakly expressed in <cell_line>H9 cells</cell_line> and not detectable in <cell_line>CEM cells</cell_line> .


After stimulation by PMA , RXR alpha expression increased in H9 and U937 cells but not in CEM cells .

After stimulation by PMA , <protein>RXR alpha</protein> expression increased in <cell_line>H9</cell_line> and <cell_line>U937 cells</cell_line> but not in <cell_line>CEM cells</cell_line> .


Human RAR alpha expression was unchanged in H9 and CEM cells , and elevated in U937 cells , after PMA stimulation .

Human RAR alpha expression was unchanged in <cell_line>H9</cell_line> and <cell_line>CEM cells</cell_line> , and elevated in <cell_line>U937 cells</cell_line> , after PMA stimulation .


CONCLUSION : The effect of RA on HIV-1 expression was cell-type-dependent and partially correlated with cellular expression of RARs .

CONCLUSION : The effect of RA on HIV-1 expression was cell-type-dependent and partially correlated with cellular expression of <protein>RARs</protein> .


Endogenous or exogenously administered RA may have a significant role in HIV regulation .


Functions of glutathione and glutathione disulfide in immunology and immunopathology .


Even a moderate increase in the cellular cysteine supply elevates the intracellular glutathione ( GSH ) and glutathione disulfide ( GSSG ) levels and potentiates immunological functions of lymphocytes in vitro .

Even a moderate increase in the cellular cysteine supply elevates the intracellular glutathione ( GSH ) and glutathione disulfide ( GSSG ) levels and potentiates immunological functions of <cell_type>lymphocytes</cell_type> in vitro .


At low GSSG levels , T cells can not optimally activate the immunologically important transcription factor NF kappa B , whereas high GSSG levels inhibit the DNA binding activity of NF kappa B .

At low GSSG levels , <cell_type>T cells</cell_type> can not optimally activate the immunologically important <protein>transcription factor</protein> <protein>NF kappa B</protein> , whereas high GSSG levels inhibit the DNA binding activity of <protein>NF kappa B</protein> .


The effects of GSSG are antagonized by reduced thioredoxin ( TRX ) .

The effects of GSSG are antagonized by reduced <protein>thioredoxin</protein> ( <protein>TRX</protein> ) .


As the protein tyrosine kinase activities p56lck and p59fyn are activated in intact cells by hydrogen peroxide , they are likely targets for GSSG action .

As the <protein>protein tyrosine kinase</protein> activities <protein>p56lck</protein> and <protein>p59fyn</protein> are activated in intact cells by hydrogen peroxide , they are likely targets for GSSG action .


These redox-regulated enzymes trigger signal cascades for NF kappa B activation and transduce signals from the T cell antigen receptor , from CD4 and CD8 molecules , and from the IL-2 receptor beta-chain .

These <protein>redox-regulated enzymes</protein> trigger signal cascades for <protein>NF kappa B</protein> activation and transduce signals from the <protein>T cell antigen receptor</protein> , from <protein>CD4</protein> and <protein>CD8 molecules</protein> , and from the <protein>IL-2 receptor beta-chain</protein> .


The effector phase of cytotoxic T cell responses and IL-2 -dependent functions are inhibited even by a partial depletion of the intracellular GSH pool .

The effector phase of <cell_type>cytotoxic T cell</cell_type> responses and <protein>IL-2</protein> -dependent functions are inhibited even by a partial depletion of the intracellular GSH pool .


As signal transduction is facilitated by prooxidant conditions , we propose that the well-known immunological consequences of GSH depletion ultimately may be results of the accompanying GSSG deficiency .


As HIV-infected patients and SIV-infected rhesus macaques have , on the average , significantly decreased plasma cyst ( e ) ine and intracellular GSH levels , we also hypothesize that AIDS may be the consequence of a GSSG deficiency as well .


T cells from renal cell carcinoma patients exhibit an abnormal pattern of kappa B-specific DNA-binding activity : a preliminary report .

<cell_type>T cells</cell_type> from renal cell carcinoma patients exhibit an abnormal pattern of kappa B-specific DNA-binding activity : a preliminary report .


Recent data suggest that the poor induction of a T-cell response to human renal cell carcinoma ( RCC ) may be related to alterations in signal transduction pathways .


We report that T cells from RCC patients have two alterations in kappa B motif-specific DNA-binding activity .

We report that <cell_type>T cells</cell_type> from RCC patients have two alterations in kappa B motif-specific DNA-binding activity .


The first alteration involves the constitutive expression of substantial kappa B-binding activity in nuclear extracts , which was observed in the electrophoretic mobility shift assay .


The magnitude of kappa B activity in unstimulated patient T cells was similar to that observed in T cells from normal individuals that had been activated in vitro .

The magnitude of kappa B activity in <cell_type>unstimulated patient T cells</cell_type> was similar to that observed in <cell_type>T cells</cell_type> from normal individuals that had been activated in vitro .


On the basis of Western blotting experiments using antibodies to kappa B/Rel family proteins , the kappa B-binding activity constitutively expressed in T cells from RCC patients is composed mostly of the NF-kappa B1 ( p50 ) subunit .

On the basis of Western blotting experiments using <protein>antibodies</protein> to <protein>kappa B/Rel family proteins</protein> , the kappa B-binding activity constitutively expressed in <cell_type>T cells</cell_type> from RCC patients is composed mostly of the <protein>NF-kappa B1 ( p50 ) subunit</protein> .


The second abnormality in kappa B-binding activity in T cells from these patients is that RelA , a member of the Rel homology family which is part of the normal NF-kappa B complex , was not induced in the nucleus following activation .

The second abnormality in kappa B-binding activity in <cell_type>T cells</cell_type> from these patients is that <protein>RelA</protein> , a member of the <protein>Rel homology family</protein> which is part of the normal <protein>NF-kappa B complex</protein> , was not induced in the nucleus following activation .


Western blotting analysis did not detect any RelA in nuclear extracts either before or after stimulation of T cells .

Western blotting analysis did not detect any <protein>RelA</protein> in nuclear extracts either before or after stimulation of <cell_type>T cells</cell_type> .


The altered kappa B-binding activity in T cells from RCC patients may impair their capacity to respond normally to various stimuli .

The altered kappa B-binding activity in <cell_type>T cells</cell_type> from RCC patients may impair their capacity to respond normally to various stimuli .


Hemoglobin switching in humans is accompanied by changes in the ratio of the transcription factors , GATA-1 and SP1 .

<protein>Hemoglobin</protein> switching in humans is accompanied by changes in the ratio of the <protein>transcription factors</protein> , <protein>GATA-1</protein> and <protein>SP1</protein> .


BACKGROUND : Understanding the mechanism of developmental regulation of hemoglobin switching has scientific as well as clinical relevance because of the influence of fetal hemoglobin ( HbF ) production in adulthood on the clinical manifestation of thalassemia and sickle cell anemia .

BACKGROUND : Understanding the mechanism of developmental regulation of <protein>hemoglobin</protein> switching has scientific as well as clinical relevance because of the influence of fetal <protein>hemoglobin</protein> ( <protein>HbF</protein> ) production in adulthood on the clinical manifestation of thalassemia and sickle cell anemia .


We have previously found that the normal developmental patterns of globin gene expression are recapitulated in an experimental system of primary cultures that support differentiation of erythroid progenitors .

We have previously found that the normal developmental patterns of <dna>globin gene</dna> expression are recapitulated in an experimental system of <cell_line>primary cultures</cell_line> that support differentiation of <cell_type>erythroid progenitors</cell_type> .


We further found that high activities of the transcriptional activators , GATA-1 and SP1 , are associated with normal adult erythroid differentiation .

We further found that high activities of the <protein>transcriptional activators</protein> , <protein>GATA-1</protein> and <protein>SP1</protein> , are associated with normal adult erythroid differentiation .


MATERIALS AND METHODS : In the present work , we have studied , the activities of GATA-1 and SP1 during differentiation of cultured erythroid progenitors derived from cord blood and from fetal livers , as well as from beta zero-thalassemia patients .

MATERIALS AND METHODS : In the present work , we have studied , the activities of <protein>GATA-1</protein> and <protein>SP1</protein> during differentiation of cultured <cell_type>erythroid progenitors</cell_type> derived from cord blood and from fetal livers , as well as from beta zero-thalassemia patients .


RESULTS : The results showed high GATA-1 binding activity and very low SP1 activity in the fetal liver cultures .

RESULTS : The results showed high <protein>GATA-1</protein> binding activity and very low <protein>SP1</protein> activity in the <cell_line>fetal liver cultures</cell_line> .


This pattern was in contrast to cultures derived from normal adult peripheral blood , in which both GATA-1 and SP1 activities were high .

This pattern was in contrast to cultures derived from normal adult peripheral blood , in which both <protein>GATA-1</protein> and <protein>SP1</protein> activities were high .


Cord blood cultures showed an additive combination of `` adult '' and `` fetal '' patterns .


The progenitors derived from a beta zero-thalassemia patient with high HbF production showed `` fetal '' pattern .

The progenitors derived from a beta zero-thalassemia patient with high <protein>HbF</protein> production showed `` fetal '' pattern .


On the other hand , in cultures of 2 beta zero-thalassemia patients without high HbF , `` adult '' pattern was observed .

On the other hand , in cultures of 2 beta zero-thalassemia patients without high <protein>HbF</protein> , `` adult '' pattern was observed .


CONCLUSIONS : In the present work , we show that human fetal and adult erythroid progenitors are distinct in their transcription factors , and that the commitment to fetal or adult program occurs at a very early differentiation stage .

CONCLUSIONS : In the present work , we show that <cell_type>human fetal and adult erythroid progenitors</cell_type> are distinct in their <protein>transcription factors</protein> , and that the commitment to fetal or adult program occurs at a very early differentiation stage .


Our studies also demonstrate that under anemic stress , recruitment of fetal progenitors may occur in adulthood .

Our studies also demonstrate that under anemic stress , recruitment of <cell_type>fetal progenitors</cell_type> may occur in adulthood .


Transcription-independent turnover of I kappa B alpha during monocyte adherence : implications for a translational component regulating I kappa B alpha/MAD-3 mRNA levels .

Transcription-independent turnover of <protein>I kappa B alpha</protein> during monocyte adherence : implications for a translational component regulating <rna>I kappa B alpha/MAD-3 mRNA</rna> levels .


We identified I kappa B alpha/MAD-3 as an immediate-early gene in human monocytes that is expressed in response to a variety of signals , including adhesion , lipopolysaccharide , and phorbol myristate acetate .

We identified <protein>I kappa B alpha/MAD-3</protein> as an <dna>immediate-early gene</dna> in <cell_type>human monocytes</cell_type> that is expressed in response to a variety of signals , including adhesion , lipopolysaccharide , and phorbol myristate acetate .


Within 5 min of monocyte adhesion , the level of the I kappa B alpha protein is markedly diminished but is rapidly replaced in a cycloheximide-sensitive manner within 20 min .

Within 5 min of monocyte adhesion , the level of the <protein>I kappa B alpha protein</protein> is markedly diminished but is rapidly replaced in a cycloheximide-sensitive manner within 20 min .


Accompanying the rapid turnover of the I kappa B alpha protein is simultaneous translocation of NF-kappa B-related transcription factors to nuclei of adhered monocytes .

Accompanying the rapid turnover of the <protein>I kappa B alpha protein</protein> is simultaneous translocation of <protein>NF-kappa B-related transcription factors</protein> to nuclei of adhered monocytes .


The demonstration that NF-kappa B can regulate I kappa B alpha/MAD-3 gene transcription in other cell types suggested that the rapid increase in steady-state I kappa B alpha/MAD-3 mRNA levels we observed within 30 min of monocyte adherence would result from NF-kappa B -dependent transcriptional stimulation of the I kappa B alpha/MAD-3 gene .

The demonstration that <protein>NF-kappa B</protein> can regulate <dna>I kappa B alpha/MAD-3 gene</dna> transcription in other cell types suggested that the rapid increase in steady-state <rna>I kappa B alpha/MAD-3 mRNA</rna> levels we observed within 30 min of monocyte adherence would result from <protein>NF-kappa B</protein> -dependent transcriptional stimulation of the <dna>I kappa B alpha/MAD-3 gene</dna> .


Nuclear run-on analyses indicated that , instead , while several immediate-early cytokine genes , such as the interleukin 1 beta ( IL-1 beta ) gene , were transcriptionally activated during monocyte adhesion , the rate of I kappa B alpha/MAD-3 gene transcription remained constant .

Nuclear run-on analyses indicated that , instead , while several <dna>immediate-early cytokine genes</dna> , such as the <dna>interleukin 1 beta ( IL-1 beta ) gene</dna> , were transcriptionally activated during monocyte adhesion , the rate of <dna>I kappa B alpha/MAD-3 gene</dna> transcription remained constant .


The adherence-dependent increase in I kappa B alpha/MAD-3 mRNA levels was also not a consequence of mRNA stabilization events .

The adherence-dependent increase in <rna>I kappa B alpha/MAD-3 mRNA</rna> levels was also not a consequence of <rna>mRNA</rna> stabilization events .


Interestingly , while increases in both IL-1 beta and I kappa B alpha/MAD-3 mRNA levels were detected in nuclei of adherent monocytes , cytoplasmic levels of IL-1 beta mRNA increased during adherence whereas those of I kappa B alpha/MAD-3 mRNA did not .

Interestingly , while increases in both IL-1 beta and I kappa B alpha/MAD-3 mRNA levels were detected in nuclei of <cell_type>adherent monocytes</cell_type> , cytoplasmic levels of <rna>IL-1 beta mRNA</rna> increased during adherence whereas those of <rna>I kappa B alpha/MAD-3 mRNA</rna> did not .


Taken together , our data suggest that two interactive mechanisms regulate monocytic I kappa B alpha/MAD-3 mRNA levels .

Taken together , our data suggest that two interactive mechanisms regulate <rna>monocytic I kappa B alpha/MAD-3 mRNA</rna> levels .


We propose that adherent monocytes regulate nuclear processing ( or decay ) of I kappa B alpha/MAD-3 mRNA , thereby increasing mRNA levels without stimulating I kappa B alpha/MAD-3 gene transcription .

We propose that <cell_type>adherent monocytes</cell_type> regulate nuclear processing ( or decay ) of <rna>I kappa B alpha/MAD-3 mRNA</rna> , thereby increasing mRNA levels without stimulating <dna>I kappa B alpha/MAD-3 gene</dna> transcription .


Moreover , since inhibition of protein synthesis leads to accumulation of I kappa B alpha/MAD-3 mRNA without stimulating I kappa B alpha/MAD-3 gene transcription , we suggest that low cytoplasmic levels of I kappa B alpha/MAD-3 mRNA are maintained by a translation-dependent degradation mechanism .

Moreover , since inhibition of protein synthesis leads to accumulation of <rna>I kappa B alpha/MAD-3 mRNA</rna> without stimulating <dna>I kappa B alpha/MAD-3 gene</dna> transcription , we suggest that low cytoplasmic levels of <rna>I kappa B alpha/MAD-3 mRNA</rna> are maintained by a translation-dependent degradation mechanism .


Distinct roles of the molecular chaperone hsp90 in modulating dioxin receptor function via the basic helix-loop-helix and PAS domains .

Distinct roles of the molecular <protein>chaperone hsp90</protein> in modulating dioxin receptor function via the basic <protein>helix-loop-helix</protein> and <protein>PAS domains</protein> .


The intracellular dioxin receptor mediates signal transduction by dioxin and functions as a ligand-activated transcription factor .

The <protein>intracellular dioxin receptor</protein> mediates signal transduction by dioxin and functions as a <protein>ligand-activated transcription factor</protein> .


It contains a basic helix-loop-helix ( bHLH ) motif contiguous with a Per-Arnt-Sim ( PAS ) homology region .

It contains a <protein>basic helix-loop-helix ( bHLH ) motif</protein> contiguous with a <protein>Per-Arnt-Sim ( PAS ) homology region</protein> .


In extracts from nonstimulated cells the receptor is recovered in an inducible cytoplasmic form associated with the 90-kDa heat shock protein ( hsp90 ) , a molecular chaperone .

In extracts from nonstimulated cells the receptor is recovered in an inducible cytoplasmic form associated with the <protein>90-kDa heat shock protein</protein> ( <protein>hsp90</protein> ) , a <protein>molecular chaperone</protein> .


We have reconstituted ligand-dependent activation of the receptor to a DNA-binding form by using the dioxin receptor and its bHLH-PAS partner factor Arnt expressed by in vitro translation in reticulocyte lysate .

We have reconstituted ligand-dependent activation of the receptor to a <protein>DNA-binding form</protein> by using the <protein>dioxin receptor</protein> and its <protein>bHLH-PAS partner factor</protein> <protein>Arnt</protein> expressed by in vitro translation in reticulocyte lysate .


Deletion of the PAS domain of the receptor resulted in constitutive dimerization with Arnt .

Deletion of the <protein>PAS domain</protein> of the receptor resulted in constitutive dimerization with <protein>Arnt</protein> .


In contrast , this receptor mutant showed low levels of xenobiotic response element-binding activity , indicating that the PAS domain may be important for DNA-binding affinity and/or specificity of the receptor .

In contrast , this receptor mutant showed low levels of xenobiotic response element-binding activity , indicating that the <protein>PAS domain</protein> may be important for DNA-binding affinity and/or specificity of the receptor .


It was not possible to reconstitute dioxin receptor function with proteins expressed in wheat germ lysate .

It was not possible to reconstitute <protein>dioxin receptor</protein> function with proteins expressed in wheat germ lysate .


In line with these observations , reticulocyte lysate but not wheat germ lysate promoted the association of de novo synthesized dioxin receptor with hsp90 .

In line with these observations , reticulocyte lysate but not wheat germ lysate promoted the association of de novo synthesized <protein>dioxin receptor</protein> with <protein>hsp90</protein> .


At least two distinct domains of the receptor mediated interaction with hsp90 : the ligand-binding domain located within the PAS region and , surprisingly , the bHLH domain .

At least two distinct domains of the receptor mediated interaction with <protein>hsp90</protein> : the <protein>ligand-binding domain</protein> located within the <protein>PAS region</protein> and , surprisingly , the <protein>bHLH domain</protein> .


Whereas ligand-binding activity correlated with association with hsp90 , bHLH- hsp90 interaction appeared to be important for DNA-binding activity but not for dimerization of the receptor .

Whereas ligand-binding activity correlated with association with <protein>hsp90</protein> , bHLH- <protein>hsp90</protein> interaction appeared to be important for DNA-binding activity but not for dimerization of the receptor .


Several distinct roles for hsp90 in modulating dioxin receptor function are therefore likely : correct folding of the ligand-binding domain , interference with Arnt heterodimerization , and folding of a DNA-binding conformation of the bHLH domain .

Several distinct roles for <protein>hsp90</protein> in modulating <protein>dioxin receptor</protein> function are therefore likely : correct folding of the ligand-binding domain , interference with <protein>Arnt</protein> heterodimerization , and folding of a DNA-binding conformation of the <protein>bHLH domain</protein> .


Thus , the dioxin receptor system provides a complex and interesting model of the regulation of transcription factors by hsp90 .

Thus , the <protein>dioxin receptor</protein> system provides a complex and interesting model of the regulation of <protein>transcription factors</protein> by <protein>hsp90</protein> .


Induction of transcription factors in human T lymphocytes by aspirin-like drugs .

Induction of <protein>transcription factors</protein> in <cell_type>human T lymphocytes</cell_type> by aspirin-like drugs .


Aspirin-like drugs ( ALD ) induce calcium mobilization , an essential component of T cell activation , but do not induce the biosynthesis of IL-2 .

Aspirin-like drugs ( ALD ) induce calcium mobilization , an essential component of T cell activation , but do not induce the biosynthesis of <protein>IL-2</protein> .


To understand the extent to which ALD may mimic mitogenic stimulation , we studied cytoplasmic and nuclear signaling steps in ALD-treated T cells .

To understand the extent to which ALD may mimic mitogenic stimulation , we studied cytoplasmic and nuclear signaling steps in <cell_line>ALD-treated T cells</cell_line> .


We found that ALD induce a transient activation of protein kinase ( PKC ) but have no effect ( in comparison to anti-CD3 antibodies ) on protein tyrosine phosphorylation nor on PCL gamma 1 tyrosine phosphorylation .

We found that ALD induce a transient activation of <protein>protein kinase</protein> ( <protein>PKC</protein> ) but have no effect ( in comparison to <protein>anti-CD3 antibodies</protein> ) on protein tyrosine phosphorylation nor on PCL gamma 1 tyrosine phosphorylation .


ALD-induced calcium mobilization and PKC activation are independent of tyrosine protein kinase activity as shown by the lack of effect of herbimycin , a tyrosine-protein kinase -specific inhibitor .

ALD-induced calcium mobilization and <protein>PKC</protein> activation are independent of <protein>tyrosine protein kinase</protein> activity as shown by the lack of effect of herbimycin , a <protein>tyrosine-protein kinase</protein> -specific inhibitor .


Although we detected no IL-2 mRNA in ALD-treated cells , the nuclei of these cells contain proteins capable of binding to three regulatory sequences in the IL-2 promoter region : NFAT , NF kappa B , and AP-1 .

Although we detected no <rna>IL-2 mRNA</rna> in <cell_line>ALD-treated cells</cell_line> , the nuclei of these cells contain proteins capable of binding to three <dna>regulatory sequences</dna> in the <dna>IL-2 promoter region</dna> : <protein>NFAT</protein> , <protein>NF kappa B</protein> , and <protein>AP-1</protein> .


These binding activities are expressed only in activated T cells .

These binding activities are expressed only in <cell_type>activated T cells</cell_type> .


The expression of AP-1 depended on calcium mobilization and PKC activation .

The expression of <protein>AP-1</protein> depended on calcium mobilization and <protein>PKC</protein> activation .


These data suggest that ALD cause transient but significant changes in T cell transmembrane signaling , although some events induced by stimulation with anti-CD3 antibodies are not induced by ALD .

These data suggest that ALD cause transient but significant changes in T cell transmembrane signaling , although some events induced by stimulation with <protein>anti-CD3 antibodies</protein> are not induced by ALD .


The signal is transmitted to the nucleus and induces DNA-binding activity by several transcription factors .

The signal is transmitted to the nucleus and induces DNA-binding activity by several <protein>transcription factors</protein> .


However , the ALD stimulus is not capable of causing complete T cell activation .


Protein kinase C is not a downstream effector of p21ras in activated T cells .

<protein>Protein kinase C</protein> is not a downstream effector of <dna>p21ras</dna> in <cell_type>activated T cells</cell_type> .


The aim of this present study was to investigate the role of protein kinase C ( PKC ) , downstream of p21ras , in activating interleukin-2 ( IL-2 ) gene expression .

The aim of this present study was to investigate the role of <protein>protein kinase C</protein> ( <protein>PKC</protein> ) , downstream of <dna>p21ras</dna> , in activating <dna>interleukin-2 ( IL-2 ) gene</dna> expression .


It has been reported that PKC is an effector of p21ras in T cells .

It has been reported that <protein>PKC</protein> is an <protein>effector</protein> of <dna>p21ras</dna> in <cell_type>T cells</cell_type> .


Data is presented , using the potent and selective PKC inhibitor Ro 31-8425 and transient expression of a constitutively active ras mutant , which clearly shows that PKC is not downstream of p21ras in the induction of NF-AT and AP-1 transcriptional activity and in the expression of IL-2 in human Jurkat T cells .

Data is presented , using the potent and selective <protein>PKC</protein> inhibitor Ro 31-8425 and transient expression of a constitutively active ras mutant , which clearly shows that <protein>PKC</protein> is not downstream of <dna>p21ras</dna> in the induction of <protein>NF-AT</protein> and <protein>AP-1</protein> transcriptional activity and in the expression of <protein>IL-2</protein> in <cell_line>human Jurkat T cells</cell_line> .


Reporter gene experiments demonstrated that NF-kappa B transcriptional activity is not affected by expression of activated p21ras .

Reporter gene experiments demonstrated that <protein>NF-kappa B</protein> transcriptional activity is not affected by expression of activated <dna>p21ras</dna> .


The signaling pathways involving PKC activation , calcium mobilization and ras activation combine to provide the necessary components for production of IL-2 during T cell activation .

The signaling pathways involving <protein>PKC</protein> activation , calcium mobilization and <protein>ras</protein> activation combine to provide the necessary components for production of <protein>IL-2</protein> during T cell activation .


The C-terminus of the B cell activator Oct-2 functions as an activation domain in yeast .

The <protein>C-terminus</protein> of the <protein>B cell activator</protein> <protein>Oct-2</protein> functions as an <protein>activation domain</protein> in yeast .


Oct-1 and Oct-2 are human transcriptional activators that bind to the same DNA element but activate distinct sets of genes .

<protein>Oct-1</protein> and <protein>Oct-2</protein> are human <protein>transcriptional activators</protein> that bind to the same DNA element but activate distinct sets of genes .


We expressed these factors in S. cerevisiae and observed greater than 5-fold stimulation of a lacZ reporter gene only with Oct-2 .

We expressed these factors in S. cerevisiae and observed greater than 5-fold stimulation of a <dna>lacZ reporter gene</dna> only with <protein>Oct-2</protein> .


Transfer of the Oct-2 C-terminal domain onto either Oct-1 ( Oct1.2 ) or a nonactivating DNA-binding domain from GAL4 created activators capable of greater than 15 and 10-fold stimulation of activity , respectively .

Transfer of the <protein>Oct-2 C-terminal domain</protein> onto either <protein>Oct-1</protein> ( <protein>Oct1.2</protein> ) or a <protein>nonactivating DNA-binding domain</protein> from <protein>GAL4</protein> created activators capable of greater than 15 and 10-fold stimulation of activity , respectively .


Thus , the C-terminus of Oct-2 is sufficient to confer activation potential to nonactive DNA-binding fragments in yeast .

Thus , the <protein>C-terminus</protein> of <protein>Oct-2</protein> is sufficient to confer activation potential to nonactive <dna>DNA-binding fragments</dna> in yeast .


Glucocorticoid-induced apoptosis of lymphoid cells .

Glucocorticoid-induced apoptosis of <cell_type>lymphoid cells</cell_type> .


The induction of cell death in lymphoid cells by glucocorticoids is one of the earliest and most thoroughly studied models of apoptosis .

The induction of cell death in <cell_type>lymphoid cells</cell_type> by glucocorticoids is one of the earliest and most thoroughly studied models of apoptosis .


Although the exact mechanism by which apoptosis occurs in lymphocytes is unknown many biochemical and molecular changes have been shown to occur in these cells in response to glucocorticoids .

Although the exact mechanism by which apoptosis occurs in <cell_type>lymphocytes</cell_type> is unknown many biochemical and molecular changes have been shown to occur in these cells in response to glucocorticoids .


The role of chromatin degradation and endonucleases in the apoptotic process has been closely studied , as well as the involvement of several oncogenes in glucocorticoid-induced cell lysis .

The role of <dna>chromatin</dna> degradation and <protein>endonucleases</protein> in the apoptotic process has been closely studied , as well as the involvement of several <dna>oncogenes</dna> in glucocorticoid-induced cell lysis .


In addition , the clinical importance of glucocorticoid-induced apoptosis in the treatment of lymphoid neoplasms has recently received increased attention .


Interleukin-2 induces tyrosine phosphorylation and nuclear translocation of stat3 in human T lymphocytes .

<protein>Interleukin-2</protein> induces tyrosine phosphorylation and nuclear translocation of <protein>stat3</protein> in <cell_type>human T lymphocytes</cell_type> .


An early biochemical event associated with T cell activation through the interleukin-2 receptor ( IL-2R ) is tyrosine phosphorylation of several intracellular substrates .

An early biochemical event associated with T cell activation through the <protein>interleukin-2 receptor</protein> ( <protein>IL-2R</protein> ) is tyrosine phosphorylation of several intracellular substrates .


The exact mechanism by which IL-2 regulates transcription of different genes is presently unknown .

The exact mechanism by which <protein>IL-2</protein> regulates transcription of different genes is presently unknown .


Here , we report that stimulation through the IL-2R induced tyrosine phosphorylation and subsequent nuclear translocation of stat3 , a newly identified member of the signal transducers and activators of transcription ( STAT ) family of proteins .

Here , we report that stimulation through the <protein>IL-2R</protein> induced tyrosine phosphorylation and subsequent nuclear translocation of <protein>stat3</protein> , a newly identified member of the <protein>signal transducers and activators of transcription ( STAT ) family</protein> of proteins .


In contrast , stat1 proteins were not tyrosine phosphorylated after IL-2 ligation , whereas tyrosine-phosphorylated stat1 proteins ( 91 and 84 kDa proteins ) were translocated to the nucleus following interferon-gamma treatment of HeLa cells .

In contrast , stat1 proteins were not tyrosine phosphorylated after <protein>IL-2</protein> ligation , whereas <protein>tyrosine-phosphorylated stat1 proteins</protein> ( <protein>91 and 84 kDa proteins</protein> ) were translocated to the nucleus following <protein>interferon-gamma</protein> treatment of <cell_line>HeLa cells</cell_line> .


Apart from stat3 , another cytoplasmic protein was tyrosine phosphorylated and subsequently translocated to the nucleus in response to IL-2 .

Apart from <protein>stat3</protein> , another <protein>cytoplasmic protein</protein> was tyrosine phosphorylated and subsequently translocated to the nucleus in response to <protein>IL-2</protein> .


This protein had an apparent molecular mass of 84 kDa and was not recognized by stat3 or stat1 mAb or antisera .

This protein had an apparent molecular mass of 84 kDa and was not recognized by <protein>stat3</protein> or <protein>stat1 mAb</protein> or antisera .


Since IL-2 induced nuclear translocation of the 84 kDa protein and stat3 followed identical kinetics , p84 is a candidate for a new , yet undefined , member of the STAT family .

Since <protein>IL-2</protein> induced nuclear translocation of the <protein>84 kDa protein</protein> and <protein>stat3</protein> followed identical kinetics , <protein>p84</protein> is a candidate for a new , yet undefined , member of the STAT family .


Taken together , we report that IL-2 induces tyrosine phosphorylation and subsequent nuclear translocation of stat3 and an as yet undefined 84-kDa protein in antigen-specific human T cell lines .

Taken together , we report that <protein>IL-2</protein> induces tyrosine phosphorylation and subsequent nuclear translocation of <protein>stat3</protein> and an as yet undefined <protein>84-kDa protein</protein> in <cell_line>antigen-specific human T cell lines</cell_line> .


Human T-cell leukemia virus type I Tax activation of NF-kappa B/Rel involves phosphorylation and degradation of I kappa B alpha and RelA ( p65 ) -mediated induction of the c-rel gene .

Human T-cell leukemia virus type I <protein>Tax</protein> activation of <protein>NF-kappa B/Rel</protein> involves phosphorylation and degradation of <protein>I kappa B alpha</protein> and <protein>RelA</protein> ( <protein>p65</protein> ) -mediated induction of the <dna>c-rel gene</dna> .


The tax gene product of human T-cell leukemia virus type I ( HTLV-I ) is a potent transcriptional activator that both stimulates viral gene expression and activates an array of cellular genes involved in T-cell growth .

The tax gene product of human T-cell leukemia virus type I ( HTLV-I ) is a potent <protein>transcriptional activator</protein> that both stimulates viral gene expression and activates an array of <dna>cellular genes</dna> involved in <cell_type>T-cell</cell_type> growth .


Tax acts indirectly by inducing or modifying the action of various host transcription factors , including members of the NF-kappa B/Rel family of enhancer-binding proteins .

Tax acts indirectly by inducing or modifying the action of various host <protein>transcription factors</protein> , including members of the <protein>NF-kappa B/Rel family</protein> of <protein>enhancer-binding proteins</protein> .


In resting T cells , many of these NF-kappa B/Rel factors are sequestered in the cytoplasm by various ankyrin-rich inhibitory proteins , including I kappa B alpha .

In <cell_type>resting T cells</cell_type> , many of these <protein>NF-kappa B/Rel factors</protein> are sequestered in the cytoplasm by various <protein>ankyrin-rich inhibitory proteins</protein> , including <protein>I kappa B alpha</protein> .


HTLV-I Tax expression leads to the constitutive nuclear expression of biologically active NF-kappa B and c-Rel complexes ; however , the biochemical mechanism ( s ) underlying this response remains poorly understood .

<protein>HTLV-I Tax</protein> expression leads to the constitutive nuclear expression of biologically active <protein>NF-kappa B</protein> and <protein>c-Rel complexes</protein> ; however , the biochemical mechanism ( s ) underlying this response remains poorly understood .


In this study , we demonstrate that Tax -stimulated nuclear expression of NF-kappa B in both HTLV-I-infected and Tax-transfected human T cells is associated with the phosphorylation and rapid proteolytic degradation of I kappa B alpha .

In this study , we demonstrate that <protein>Tax</protein> -stimulated nuclear expression of <protein>NF-kappa B</protein> in both <cell_line>HTLV-I-infected and Tax-transfected human T cells</cell_line> is associated with the phosphorylation and rapid proteolytic degradation of <protein>I kappa B alpha</protein> .


In contrast to prior in vitro studies , at least a fraction of the phosphorylated form of I kappa B alpha remains physically associated with the NF-kappa B complex in vivo but is subject to rapid degradation , thereby promoting the nuclear translocation of the active NF-kappa B complex .

In contrast to prior in vitro studies , at least a fraction of the phosphorylated form of <protein>I kappa B alpha</protein> remains physically associated with the <protein>NF-kappa B complex</protein> in vivo but is subject to rapid degradation , thereby promoting the nuclear translocation of the active <protein>NF-kappa B complex</protein> .


We further demonstrate that Tax induction of nuclear c-Rel expression is activated by the RelA ( p65 ) subunit of NF-kappa B , which activates transcription of the c-rel gene through an intrinsic kappa B enhancer element .

We further demonstrate that <protein>Tax</protein> induction of nuclear <protein>c-Rel</protein> expression is activated by the <protein>RelA</protein> ( <protein>p65</protein> ) subunit of <protein>NF-kappa B</protein> , which activates transcription of the <dna>c-rel gene</dna> through an intrinsic <dna>kappa B enhancer element</dna> .


In normal cells , the subsequent accumulation of nuclear c-Rel acts to inhibit its own continued production , indicating the presence of an autoregulatory loop .

In normal cells , the subsequent accumulation of <protein>nuclear c-Rel</protein> acts to inhibit its own continued production , indicating the presence of an autoregulatory loop .


( ABSTRACT TRUNCATED AT 250 WORDS )


Activation of NF-kappa B in vivo is regulated by multiple phosphorylations .

Activation of <protein>NF-kappa B</protein> in vivo is regulated by multiple phosphorylations .


The activation of nuclear factor kappa B ( NF-kappa B ) in intact cells is mechanistically not well understood .

The activation of <protein>nuclear factor kappa B</protein> ( <protein>NF-kappa B</protein> ) in intact cells is mechanistically not well understood .


Therefore we investigated the modifications imposed on NF-kappa B/I kappa B components following stimulation and show that the final step of NF-kappa B induction in vivo involves phosphorylation of several members of the NF-kappa B/I kappa B protein families .

Therefore we investigated the modifications imposed on <protein>NF-kappa B/I kappa B components</protein> following stimulation and show that the final step of <protein>NF-kappa B</protein> induction in vivo involves phosphorylation of several members of the <protein>NF-kappa B/I kappa B protein families</protein> .


In HeLa cells as well as in B cells , TNF-alpha rapidly induced nuclear translocation primarily of p50-p65 , but not of c-rel .

In <cell_line>HeLa cells</cell_line> as well as in <cell_type>B cells</cell_type> , TNF-alpha rapidly induced nuclear translocation primarily of <protein>p50-p65</protein> , but not of <protein>c-rel</protein> .


Both NF-kappa B precursors and I kappa B alpha became strongly phosphorylated with the same kinetics .

Both <protein>NF-kappa B</protein> precursors and <protein>I kappa B alpha</protein> became strongly phosphorylated with the same kinetics .


In addition to the inducible phosphorylation after stimulation , B lymphocytes containing constitutive nuclear NF-kappa B revealed constitutively phosphorylated p65 and I kappa B alpha .

In addition to the inducible phosphorylation after stimulation , <cell_type>B lymphocytes</cell_type> containing constitutive nuclear <protein>NF-kappa B</protein> revealed constitutively phosphorylated <protein>p65</protein> and <protein>I kappa B alpha</protein> .


Phosphorylation was accompanied by induced processing of the precursors p100 and p105 and by degradation of I kappa B alpha .

Phosphorylation was accompanied by induced processing of the precursors <protein>p100</protein> and <protein>p105</protein> and by degradation of <protein>I kappa B alpha</protein> .


As an in vitro model we show that phosphorylation of p105 impedes its ability to interact with NF-kappa B , as has been shown before for I kappa B alpha .

As an in vitro model we show that phosphorylation of <protein>p105</protein> impedes its ability to interact with <protein>NF-kappa B</protein> , as has been shown before for <protein>I kappa B alpha</protein> .


Surprisingly , even p65 , but not c-rel , was phosphorylated after induction in vivo , suggesting that TNF-alpha selectively activates only specific NF-kappa B heteromers and that modifications regulate not only I kappa B molecules but also NF-kappa B molecules .

Surprisingly , even <protein>p65</protein> , but not <protein>c-rel</protein> , was phosphorylated after induction in vivo , suggesting that <protein>TNF-alpha</protein> selectively activates only specific <protein>NF-kappa B</protein> heteromers and that modifications regulate not only <protein>I kappa B molecules</protein> but also <protein>NF-kappa B</protein> molecules .


In fact , cellular NF-kappa B activity was phosphorylation-dependent and the DNA binding activity of p65-containing NF-kappa B was enhanced by phosphorylation in vitro .

In fact , cellular <protein>NF-kappa B</protein> activity was phosphorylation-dependent and the DNA binding activity of <protein>p65-containing NF-kappa B</protein> was enhanced by phosphorylation in vitro .


Furthermore , we found that the induction by hydrogen peroxide of NF-kappa B translocation to the nucleus , which is assumed to be triggered by reactive oxygen intermediates , also coincided with incorporation of phosphate into the same subunits that were modified after stimulation by TNF-alpha .

Furthermore , we found that the induction by hydrogen peroxide of <protein>NF-kappa B</protein> translocation to the nucleus , which is assumed to be triggered by reactive oxygen intermediates , also coincided with incorporation of phosphate into the same subunits that were modified after stimulation by <protein>TNF-alpha</protein> .


Thus , phosphorylation appears to be a general mechanism for activation of NF-kappa B in vivo .

Thus , phosphorylation appears to be a general mechanism for activation of <protein>NF-kappa B</protein> in vivo .


Treatment of HL60 cells with various combinations of retinoids and 1 alpha , 25 dihydroxyvitamin D3 results in differentiation towards neutrophils or monocytes or a failure to differentiate and apoptosis .

Treatment of <cell_line>HL60 cells</cell_line> with various combinations of retinoids and 1 alpha , 25 dihydroxyvitamin D3 results in differentiation towards <cell_type>neutrophils</cell_type> or <cell_type>monocytes</cell_type> or a failure to differentiate and apoptosis .


It is well documented that treatment of serum-grown HL60 cells with 10 ( -7 ) M all-trans retinoic acid ( all-trans RA ) induces neutrophil differentiation , whereas treatment with 10 ( -7 ) M 1 alpha , 25 dihydroxyvitamin D3 ( D3 ) induces differentiation towards monocytes .

It is well documented that treatment of <cell_line>serum-grown HL60 cells</cell_line> with 10 ( -7 ) M all-trans retinoic acid ( all-trans RA ) induces <cell_type>neutrophil</cell_type> differentiation , whereas treatment with 10 ( -7 ) M 1 alpha , 25 dihydroxyvitamin D3 ( D3 ) induces differentiation towards <cell_type>monocytes</cell_type> .


In recent investigations , using serum-free grown HL60 cells , we observed that all-trans RA , at 10 ( -7 ) M , did not induce neutrophil differentiation and that all-trans RA , at 10 ( -8 ) M , reduced the D3 concentration required for monocyte differentiation to 5 x 10 ( -9 ) M .

In recent investigations , using <cell_line>serum-free grown HL60 cells</cell_line> , we observed that all-trans RA , at 10 ( -7 ) M , did not induce neutrophil differentiation and that all-trans RA , at 10 ( -8 ) M , reduced the D3 concentration required for <cell_type>monocyte</cell_type> differentiation to 5 x 10 ( -9 ) M .


In this study , co-operative interactions between all-trans and 9-cis RA and D3 which promote neutrophil and monocyte differentiation of HL60 cells have been analysed in detail .

In this study , co-operative interactions between all-trans and 9-cis RA and D3 which promote neutrophil and monocyte differentiation of <cell_line>HL60 cells</cell_line> have been analysed in detail .


Treatment of serum-free grown HL60 cells with 5 x 10 ( -7 ) M all-trans RA or 9-cis RA resulted in sub-optimal neutrophil differentiation ( up to 25 % mature cells ) .

Treatment of <cell_line>serum-free grown HL60 cells</cell_line> with 5 x 10 ( -7 ) M all-trans RA or 9-cis RA resulted in sub-optimal neutrophil differentiation ( up to 25 % mature cells ) .


As shown for all-trans RA , 9-cis RA cooperated with D3 to promote monocyte differentiation .


Culture of HL60 cells in 5 x 10 ( -7 ) M 9-cis RA together with a wide range of concentrations of D3 resulted in promotion of neutrophil differentiation at 10 ( -15 ) -10 ( -12 ) D3 , a failure to differentiate and apoptosis at 10 ( -11 ) -10 ( -10 ) M D3 , followed by co-operativity between 9-cis RA and 5 x 10 ( -9 ) M D3 in inducing monocyte differentiation in the absence of neutrophil differentiation .

Culture of <cell_line>HL60 cells</cell_line> in 5 x 10 ( -7 ) M 9-cis RA together with a wide range of concentrations of D3 resulted in promotion of neutrophil differentiation at 10 ( -15 ) -10 ( -12 ) D3 , a failure to differentiate and apoptosis at 10 ( -11 ) -10 ( -10 ) M D3 , followed by co-operativity between 9-cis RA and 5 x 10 ( -9 ) M D3 in inducing monocyte differentiation in the absence of neutrophil differentiation .


Similar results were obtained when HL60 cells were treated with 5 x 10 ( -7 ) all-trans RA together with a wide range of concentrations of D3 .

Similar results were obtained when <cell_line>HL60 cells</cell_line> were treated with 5 x 10 ( -7 ) all-trans RA together with a wide range of concentrations of D3 .


Cross titration analyses of the effects of 9-cis RA and D3 on HL60 cell differentiation were undertaken to determine the boundaries of the concentrations of each agent , alone and in combination , that give rise to optimal neutrophil and monocyte differentiation of HL60 cells .

Cross titration analyses of the effects of 9-cis RA and D3 on <cell_line>HL60 cell</cell_line> differentiation were undertaken to determine the boundaries of the concentrations of each agent , alone and in combination , that give rise to optimal neutrophil and monocyte differentiation of <cell_line>HL60 cells</cell_line> .


The observed cooperativities between either 9-cis RA or all-trans RA and D3 have important implications for the use of combinations of these agents in differentiation therapy .


Isolation of differentially expressed sequence tags from steroid-responsive cells using mRNA differential display .

Isolation of differentially expressed sequence tags from <cell_type>steroid-responsive cells</cell_type> using <rna>mRNA</rna> differential display .


Transcriptional control of steroid-regulated gene networks by nuclear receptor proteins results in the coordinate expression of a limited number of target genes .

Transcriptional control of <dna>steroid-regulated gene networks</dna> by <protein>nuclear receptor proteins</protein> results in the coordinate expression of a limited number of <dna>target genes</dna> .


Although much is known about the structure and function of steroid receptors , relatively few cell-specific steroid-regulated genes have been isolated and characterized .


In this paper we describe results using mRNA differential display reverse transcriptase PCR ( DDPCR ) to identify and isolate short cDNA sequence tags from thymocyte and prostate cells under various hormone conditions .

In this paper we describe results using <rna>mRNA</rna> differential display <protein>reverse transcriptase</protein> PCR ( DDPCR ) to identify and isolate short cDNA sequence tags from <cell_type>thymocyte</cell_type> and <cell_type>prostate cells</cell_type> under various hormone conditions .


Using this technique we have isolated several differentially expressed sequence tags ( DESTs ) from the mouse thymocyte cell line WEHI 7.2 .

Using this technique we have isolated several <dna>differentially expressed sequence tags</dna> ( <dna>DESTs</dna> ) from the <cell_line>mouse thymocyte cell line</cell_line> <cell_line>WEHI 7.2</cell_line> .


Two of these DESTs , GIG10 and GIG18 , are rapidly induced by dexamethasone within 2 h of treatment .

Two of these <dna>DESTs</dna> , <dna>GIG10</dna> and <dna>GIG18</dna> , are rapidly induced by dexamethasone within 2 h of treatment .


GIG10 is a novel sequence and GIG18 is the mouse homologue of a human expressed sequence tag isolated from activated B lymphocytes .

<dna>GIG10</dna> is a novel sequence and <dna>GIG18</dna> is the mouse homologue of a <dna>human expressed sequence tag</dna> isolated from <cell_type>activated B lymphocytes</cell_type> .


We also used DDPCR to isolate DESTs from androgen-modulated rat ventral prostate tissue , one of which we characterized and found to correspond to the 3 ' end of prostatic spermine binding protein mRNA , a known androgen-regulated gene .

We also used DDPCR to isolate <dna>DESTs</dna> from androgen-modulated rat ventral prostate tissue , one of which we characterized and found to correspond to the <rna>3 ' end</rna> of <rna>prostatic spermine binding protein mRNA</rna> , a known <dna>androgen-regulated gene</dna> .


Modifications of the original DDPCR protocol , which we found can potentially decrease the frequency of isolating false-positive DESTs , are described and the merits of DDPCR , relative to other differential cDNA cloning strategies , are discussed .

Modifications of the original DDPCR protocol , which we found can potentially decrease the frequency of isolating false-positive <dna>DESTs</dna> , are described and the merits of DDPCR , relative to other differential cDNA cloning strategies , are discussed .


Activation of human thymocytes after infection by EBV .

Activation of <cell_type>human thymocytes</cell_type> after infection by EBV .


The discovery of EBV in certain T cell malignancies and the expression of the EBV receptor , CR2/CD21 , on a population of immature thymocytes , T lymphoblastoid cell lines , and childhood acute T lymphoblastic leukemia cells suggested that EBV-receptor interactions on T cells may be of importance .

The discovery of EBV in certain T cell malignancies and the expression of the <protein>EBV receptor</protein> , <protein>CR2/CD21</protein> , on a population of <cell_type>immature thymocytes</cell_type> , <cell_line>T lymphoblastoid cell lines</cell_line> , and <cell_type>childhood acute T lymphoblastic leukemia cells</cell_type> suggested that EBV-receptor interactions on <cell_type>T cells</cell_type> may be of importance .


We have shown that , within the thymus , a population of large , immature cells expresses CD21 .

We have shown that , within the thymus , a population of <cell_type>large , immature cells</cell_type> expresses <protein>CD21</protein> .


EBV altered the activation responses of immature thymocytes in vitro .

EBV altered the activation responses of <cell_type>immature thymocytes</cell_type> in vitro .


Triggering through CD2 is mitogenic for mature , but not immature , T cells .

Triggering through <protein>CD2</protein> is mitogenic for mature , but not immature , <cell_type>T cells</cell_type> .


However , during infection by EBV , ligation of CD2 caused thymocytes to proliferate in the absence of exogenous cytokines .

However , during infection by EBV , ligation of <protein>CD2</protein> caused <cell_type>thymocytes</cell_type> to proliferate in the absence of <protein>exogenous cytokines</protein> .


This function was a result of the interaction of EBV with its receptor , CD21 , but was caused by infection rather than surface signaling , because neither specific mAb nor the P3HR-1 strain of virus mimicked the effect of B95-8 .

This function was a result of the interaction of EBV with its receptor , <protein>CD21</protein> , but was caused by infection rather than surface signaling , because neither <protein>specific mAb</protein> nor the P3HR-1 strain of virus mimicked the effect of B95-8 .


Immature thymocytes were infected by EBV , as determined by the internalization of the viral genome and its transcriptional activity .

Immature <cell_type>thymocytes</cell_type> were infected by EBV , as determined by the internalization of the <dna>viral genome</dna> and its transcriptional activity .


Consistent with the activity of B95-8 , EBNA-2 transcripts were identified within infected thymocyte populations .

Consistent with the activity of <protein>B95-8</protein> , <rna>EBNA-2 transcripts</rna> were identified within <cell_line>infected thymocyte populations</cell_line> .


In addition , components of the viral replicative pathway were expressed during infection of thymocytes .

In addition , components of the viral replicative pathway were expressed during infection of <cell_type>thymocytes</cell_type> .


These components included transcription of BZLF-1 , an early gene that characterizes EBV-infected B cells after disruption of latency .

These components included transcription of <dna>BZLF-1</dna> , an <dna>early gene</dna> that characterizes <cell_type>EBV-infected B cells</cell_type> after disruption of latency .


A second transcript was identified as encoding the recently characterized RAZ , which also is associated with replicative infection .

A second transcript was identified as encoding the recently characterized <protein>RAZ</protein> , which also is associated with replicative infection .


The consequences of EBV infection of T cells at an early stage of differentiation may lead to failure of normal T cell repertoire development , autoimmunity , or malignancy .

The consequences of EBV infection of <cell_type>T cells</cell_type> at an early stage of differentiation may lead to failure of normal T cell repertoire development , autoimmunity , or malignancy .


Effects of intranasal glucocorticoids on endogenous glucocorticoid peripheral and central function .


Glucocorticoids are among the most potent antiinflammatory agents that can be used in the treatment of rhinitis .


Their mechanisms of action are multiple and complex and a number of reports describe significant systemic effects of locally administered glucocorticoids .


In order to evaluate the short-term systemic effects of intranasally administered glucocorticoids , 14 normal healthy subjects were treated with two doses of either budesonide ( BUD ) or fluticasone propionate ( FP ) for 2 weeks .


Before treatment , at regular intervals during the treatment , 1 week and finally 6 weeks after termination of treatment , the effects on glucocorticoid receptor ( GR ) and methallothionein ( MTIIa ) mRNA expression levels were examined in peripheral lymphocytes using a solution hybridization assay .

Before treatment , at regular intervals during the treatment , 1 week and finally 6 weeks after termination of treatment , the effects on glucocorticoid receptor ( GR ) and methallothionein ( MTIIa ) mRNA expression levels were examined in <cell_type>peripheral lymphocytes</cell_type> using a solution hybridization assay .


Serum cortisol , osteocalcin and urinary cortisol levels were also determined .


An insulin tolerance test ( ITT ) was performed at the end of the second week of treatment and at the end of the 6-week washout period with no statistically significant change in cortisol response .


In peripheral lymphocytes , GR mRNA levels were significantly down-regulated .

In <cell_type>peripheral lymphocytes</cell_type> , <rna>GR mRNA</rna> levels were significantly down-regulated .


MTIIa mRNA levels increased significantly .

<rna>MTIIa mRNA</rna> levels increased significantly .


Serum osteocalcin decreased significantly during treatment with both BUD and FP .

Serum <protein>osteocalcin</protein> decreased significantly during treatment with both BUD and FP .


Serum cortisol decreased after 1 week of treatment whereas urinary cortisol was not affected until the second week of treatment .


In conclusion , intranasal glucocorticoids at clinically recommended doses have not only significant systemic effects on adrenal function , but also have an effect on specific gene expression in peripheral lymphocytes .

In conclusion , intranasal glucocorticoids at clinically recommended doses have not only significant systemic effects on adrenal function , but also have an effect on specific gene expression in <cell_type>peripheral lymphocytes</cell_type> .


These effects are receptor-dependent , reversible , and according to serum and urinary cortisol levels and ITT , leave the hypothalamic-pituitary-adrenal function intact .


Finally , these short-term systemic effects were not associated with any of the noticeable side-effects usually observed during long-term treatment with glucocorticoids .


Biphasic control of nuclear factor-kappa B activation by the T cell receptor complex : role of tumor necrosis factor alpha .

Biphasic control of <protein>nuclear factor-kappa B</protein> activation by the <protein>T cell receptor complex</protein> : role of <protein>tumor necrosis factor alpha</protein> .


The regulation of nuclear factor ( NF ) -kappa B activation by the T cell receptor ( TcR ) /CD3 complex in primary human T cells has been studied at various times after activation .

The regulation of <protein>nuclear factor ( NF ) -kappa B</protein> activation by the <protein>T cell receptor ( TcR ) /CD3 complex</protein> in primary human <cell_type>T cells</cell_type> has been studied at various times after activation .


Only p50 NF-kappa B protein bound the kappa B element of interleukin-2 receptor ( IL-2R ) alpha chain promoter on resting T cells .

Only <protein>p50 NF-kappa B protein</protein> bound the <dna>kappa B element</dna> of <dna>interleukin-2 receptor ( IL-2R ) alpha chain promoter</dna> on resting <cell_type>T cells</cell_type> .


However , immediately after TcR/CD3 cross-linking ( after approximately 1 h ; immediate ) binding of p50.p65 heterodimers was observed .

However , immediately after <protein>TcR/CD3</protein> cross-linking ( after approximately 1 h ; immediate ) binding of <protein>p50.p65 heterodimers</protein> was observed .


p50.c-rel heterodimers were also detected bound to this sequence at early time points ( 7-16 h ; early ) , and both remained active at later time points ( 40 h ; late ) after activation .

<protein>p50.c-rel heterodimers</protein> were also detected bound to this sequence at early time points ( 7-16 h ; early ) , and both remained active at later time points ( 40 h ; late ) after activation .


This regulation takes place mainly at the level of nuclear translocation of p65 and c-rel , at immediate and early time points .

This regulation takes place mainly at the level of nuclear translocation of <protein>p65</protein> and <protein>c-rel</protein> , at immediate and early time points .


Activation also induced c-rel and p105/p50 mRNA synthesis , but not p65 mRNA whose expression was constitutive .

Activation also induced c-rel and p105/p50 mRNA synthesis , but not <rna>p65 mRNA</rna> whose expression was constitutive .


Interestingly , all those early and late events , but not the immediate ones , were inhibited by a neutralizing anti-tumor necrosis factor alpha ( TNF-alpha ) monoclonal antibody .

Interestingly , all those early and late events , but not the immediate ones , were inhibited by a neutralizing <protein>anti-tumor necrosis factor alpha ( TNF-alpha ) monoclonal antibody</protein> .


Similarly , cycloheximide prevented the p65 and c-rel translocation and consequent formation of active binding heterodimers , at early and late times .

Similarly , cycloheximide prevented the <protein>p65</protein> and <protein>c-rel</protein> translocation and consequent formation of active binding heterodimers , at early and late times .


Cyclosporin A impaired not only early and late , but also immediate events ; however , addition of TNF-alpha prevented all inhibition .

Cyclosporin A impaired not only early and late , but also immediate events ; however , addition of <protein>TNF-alpha</protein> prevented all inhibition .


These results indicate that the regulation of NF-kappa B activation during T cell activation by TcR/CD3 signals is biphasic : TcR/CD3 triggers its immediate translocation , which is transient if no TNF-alpha is present .

These results indicate that the regulation of <protein>NF-kappa B</protein> activation during T cell activation by <protein>TcR/CD3</protein> signals is biphasic : <protein>TcR/CD3</protein> triggers its immediate translocation , which is transient if no <protein>TNF-alpha</protein> is present .


TNF-alpha , therefore , emerges as the main factor responsible for a second phase of NF-kappa B regulation , controlling both translocation of p65 and c-rel , and new mRNA synthesis for c-rel and p105/p50 .

<protein>TNF-alpha</protein> , therefore , emerges as the main factor responsible for a second phase of <protein>NF-kappa B</protein> regulation , controlling both translocation of <protein>p65</protein> and <protein>c-rel</protein> , and new <rna>mRNA</rna> synthesis for <protein>c-rel</protein> and <protein>p105/p50</protein> .


The transcription factor Sp1 is required for induction of the murine GM-CSF promoter in T cells .

The <protein>transcription factor Sp1</protein> is required for induction of the <dna>murine GM-CSF promoter</dna> in <cell_type>T cells</cell_type> .


The cis-acting region , GM-kappa B/GC-box ( positions -95 and -73 ) , within the murine GM-CSF gene promoter is required for maximal induction by stimulation with phorbol-12-myristate acetate ( PMA ) and calcium ionophore ( A23187 ) in T cells .

The <dna>cis-acting region</dna> , <dna>GM-kappa B/GC-box</dna> ( <dna>positions -95</dna> and <dna>-73</dna> ) , within the <dna>murine GM-CSF gene promoter</dna> is required for maximal induction by stimulation with phorbol-12-myristate acetate ( PMA ) and calcium ionophore ( A23187 ) in <cell_type>T cells</cell_type> .


GM-kappa B defines a binding site for NF-kappa B , and GC-box defines a binding site for three ( A1 , A2 , B ) constitutive proteins .

<protein>GM-kappa B</protein> defines a binding site for <protein>NF-kappa B</protein> , and <dna>GC-box</dna> defines a binding site for three ( <protein>A1</protein> , <protein>A2</protein> , <protein>B</protein> ) <protein>constitutive proteins</protein> .


We report here that three copies of the GC-box can functionally compensate for the GM-kappa B/GC-box region , suggesting that the GC-motif can function independently of the GM-kappa B motif .

We report here that three copies of the <dna>GC-box</dna> can functionally compensate for the <dna>GM-kappa B/GC-box region</dna> , suggesting that the GC-motif can function independently of the <dna>GM-kappa B motif</dna> .


The major GC-box binding activity A1 was purified and identified as the transcription factor Sp1 .

The major <dna>GC-box</dna> binding activity <protein>A1</protein> was purified and identified as the <protein>transcription factor Sp1</protein> .


We show that depletion of Sp1 ( A1 ) from nuclear extracts specifically decreases in vitro transcription activity on GM-CSF templates .

We show that depletion of <protein>Sp1</protein> ( <protein>A1</protein> ) from nuclear extracts specifically decreases in vitro transcription activity on <protein>GM-CSF</protein> templates .


Since the human GM-CSF promoter has a base difference within the GC-box , we speculate that this may explain why the human promoter is weak and that an upstream enhancer is required for the induction of the human GM-CSF gene .

Since the <dna>human GM-CSF promoter</dna> has a base difference within the <dna>GC-box</dna> , we speculate that this may explain why the human promoter is weak and that an <dna>upstream enhancer</dna> is required for the induction of the <dna>human GM-CSF gene</dna> .


A factor that regulates the class II major histocompatibility complex gene DPA is a member of a subfamily of zinc finger proteins that includes a Drosophila developmental control protein .

A factor that regulates the <dna>class II major histocompatibility complex gene</dna> <dna>DPA</dna> is a member of a subfamily of <protein>zinc finger proteins</protein> that includes a <protein>Drosophila developmental control protein</protein> .


A novel DNA sequence element termed the J element involved in the regulated expression of class II major histocompatibility complex genes was recently described .

A novel <dna>DNA sequence element</dna> termed the <dna>J element</dna> involved in the regulated expression of <dna>class II major histocompatibility complex genes</dna> was recently described .


To study this element and its role in class II gene regulation further , a cDNA library was screened with oligonucleotide probes containing both the S element and the nearby J element of the human DPA gene .

To study this element and its role in <dna>class II gene</dna> regulation further , a <dna>cDNA library</dna> was screened with oligonucleotide probes containing both the <dna>S element</dna> and the nearby <dna>J element</dna> of the <dna>human DPA gene</dna> .


Several DNA clones were obtained by this procedure , one of which , clone 18 , is reported and characterized here .

Several <dna>DNA clones</dna> were obtained by this procedure , one of which , <dna>clone 18</dna> , is reported and characterized here .


It encodes a protein predicted to contain 688 amino acid residues , including 11 zinc finger motifs of the C2H2 type in the C-terminal region , that are Kruppel-like in the conservation of the H/C link sequence connecting them .

It encodes a protein predicted to contain 688 amino acid residues , including 11 <protein>zinc finger motifs</protein> of the <protein>C2H2 type</protein> in the <protein>C-terminal region</protein> , that are Kruppel-like in the conservation of the <protein>H/C link sequence</protein> connecting them .


The 160 N-terminal amino acids in the nonfinger region of clone 18 are highly homologous with similar regions of several other human , mouse , and Drosophila sequences , defining a subfamily of Kruppel-like zinc finger proteins termed TAB ( tramtrack [ ttk ] -associated box ) here .

The <protein>160 N-terminal amino acids</protein> in the <protein>nonfinger region</protein> of <protein>clone 18</protein> are highly homologous with similar regions of several other <dna>human , mouse , and Drosophila sequences</dna> , defining a subfamily of Kruppel-like <protein>zinc finger proteins</protein> termed <protein>TAB</protein> ( <protein>tramtrack [ ttk ] -associated box</protein> ) here .


One of the Drosophila sequences , ttk , is a developmental control gene , while a second does not contain a zinc finger region but encodes a structure important in oocyte development .

One of the <dna>Drosophila sequences</dna> , <dna>ttk</dna> , is a <dna>developmental control gene</dna> , while a second does not contain a <protein>zinc finger region</protein> but encodes a structure important in oocyte development .


An acidic activation domain is located between the N-terminal conserved region of clone 18 and its zinc fingers .

An <protein>acidic activation domain</protein> is located between the <protein>N-terminal conserved region</protein> of <protein>clone 18</protein> and its <protein>zinc fingers</protein> .


This protein appears to require both the S and J elements , which are separated by 10 bp for optimal binding .

This protein appears to require both the <dna>S and J elements</dna> , which are separated by 10 bp for optimal binding .


Antisense cDNA to clone 18 inhibited the expression of a reporter construct containing the DPA promoter , indicating its functional importance in the expression of this class II gene .

Antisense cDNA to <protein>clone 18</protein> inhibited the expression of a reporter construct containing the <dna>DPA promoter</dna> , indicating its functional importance in the expression of this <dna>class II gene</dna> .


Separation of oxidant-initiated and redox-regulated steps in the NF-kappa B signal transduction pathway .

Separation of oxidant-initiated and redox-regulated steps in the <protein>NF-kappa B</protein> signal transduction pathway .


Studies presented here show that overall NF-kappa B signal transduction begins with a parallel series of stimuli-specific pathways through which cytokines ( tumor necrosis factor alpha ) , oxidants ( hydrogen peroxide and mitomycin C ) , and phorbol ester ( phorbol 12-myristate 13-acetate ) individually initiate signaling .

Studies presented here show that overall <protein>NF-kappa B</protein> signal transduction begins with a parallel series of stimuli-specific pathways through which <protein>cytokines</protein> ( <protein>tumor necrosis factor alpha</protein> ) , oxidants ( hydrogen peroxide and mitomycin C ) , and phorbol ester ( phorbol 12-myristate 13-acetate ) individually initiate signaling .


These initial pathways culminate in a common pathway through which all of the stimulating agents ultimately signal NF-kappa B activation .

These initial pathways culminate in a common pathway through which all of the stimulating agents ultimately signal <protein>NF-kappa B</protein> activation .


We distinguish the stimuli-specific pathways by showing that the oxidative stimuli trigger NF-kappa B activation in only one of two human T-cell lines ( Wurzburg but not Jurkat ) , whereas tumor necrosis factor alpha and phorbol 12-myristate 13-acetate readily stimulate in both lines .

We distinguish the stimuli-specific pathways by showing that the oxidative stimuli trigger <protein>NF-kappa B</protein> activation in only one of two <cell_line>human T-cell lines</cell_line> ( <cell_line>Wurzburg</cell_line> but not <cell_line>Jurkat</cell_line> ) , whereas <protein>tumor necrosis factor alpha</protein> and phorbol 12-myristate 13-acetate readily stimulate in both lines .


We propose the common pathway as the simplest way of accounting for the common requirements and properties of the signaling pathway .


We include a redox-regulatory mechanism ( s ) in this common pathway to account for the previously demonstrated redox regulation of NF-kappa B activation in Jurkat cells ( in which oxidants do n't activate NF-kappa B ) ; we put tyrosine phosphorylation in the common pathway by showing that kinase activity ( inhibitable by herbimycin A and tyrphostin 47 ) is required for NF-kappa B activation by all stimuli tested in both cell lines .

We include a redox-regulatory mechanism ( s ) in this common pathway to account for the previously demonstrated redox regulation of <protein>NF-kappa B</protein> activation in <cell_line>Jurkat cells</cell_line> ( in which oxidants do n't activate <protein>NF-kappa B</protein> ) ; we put tyrosine phosphorylation in the common pathway by showing that kinase activity ( inhibitable by herbimycin A and tyrphostin 47 ) is required for <protein>NF-kappa B</protein> activation by all stimuli tested in both cell lines .


Since internal sites of oxidant production have been shown to play a key role in the cytokine -stimulated activation of NF-kappa B , and since tyrosine kinase and phosphatase activities are known to be altered by oxidants , these findings suggest that intracellular redox status controls NF-kappa B activation by regulating tyrosine phosphorylation event ( s ) within the common step of the NF-kappa B signal transduction pathway .

Since internal sites of oxidant production have been shown to play a key role in the <protein>cytokine</protein> -stimulated activation of <protein>NF-kappa B</protein> , and since tyrosine kinase and phosphatase activities are known to be altered by oxidants , these findings suggest that intracellular redox status controls <protein>NF-kappa B</protein> activation by regulating tyrosine phosphorylation event ( s ) within the common step of the <protein>NF-kappa B</protein> signal transduction pathway .


Deleted chromosome 20 from a patient with Alagille syndrome isolated in a cell hybrid through leucine transport selection : study of three candidate genes .

Deleted <dna>chromosome 20</dna> from a patient with Alagille syndrome isolated in a cell hybrid through leucine transport selection : study of three candidate genes .


Alagille syndrome ( AGS ) is a well-defined genetic entity assigned to the short arm of Chromosome ( Chr ) 20 by a series of observations of AGS patients associated with microdeletions in this region .

Alagille syndrome ( AGS ) is a well-defined genetic entity assigned to the <dna>short arm</dna> of <dna>Chromosome ( Chr ) 20</dna> by a series of observations of AGS patients associated with microdeletions in this region .


By fusing lymphoblastoid cells of an AGS patient that exhibited a microdeletion in the short arm of Chr 20 encompassing bands p11.23 to p12.3 with rodent thermosensitive mutant cells ( CHOtsH1-1 ) deficient in-leucyl-tRNA synthetase , we isolated a somatic cell hybrid segregating the deleted human Chr 20 .

By fusing <cell_type>lymphoblastoid cells</cell_type> of an AGS patient that exhibited a microdeletion in the <dna>short arm of Chr 20</dna> encompassing bands <dna>p11.23 to p12.3</dna> with <cell_line>rodent thermosensitive mutant cells</cell_line> ( <cell_line>CHOtsH1-1</cell_line> ) deficient <protein>in-leucyl-tRNA synthetase</protein> , we isolated a <cell_line>somatic cell hybrid</cell_line> segregating the deleted human <dna>Chr 20</dna> .


This hybrid clone , designated NR2 , was characterized by several methods , including PCR , with eight pairs of oligonucleotides mapped to Chr 20 : D20S5 , D20S41 , D20S42 , D20S56 , D20S57 , D20S58 , adenosine deaminase ( ADA ) , and Prion protein ( PRIP ) ; Restriction Fragment Length Polymorphism ( RFLP ) analyses with four genomic anonymous probes ( D20S5 , cD3H12 , D20S17 , D20S18 ) ; and fluorescent in situ hybridization ( FISH ) with total human DNA and D20Z1 , a sequence specific to the human Chr 20 centromere , as probes .

This hybrid clone , designated <cell_line>NR2</cell_line> , was characterized by several methods , including PCR , with eight pairs of oligonucleotides mapped to <dna>Chr 20</dna> : <dna>D20S5</dna> , <dna>D20S41</dna> , <dna>D20S42</dna> , <dna>D20S56</dna> , <dna>D20S57</dna> , <dna>D20S58</dna> , <dna>adenosine deaminase</dna> ( <protein>ADA</protein> ) , and <dna>Prion protein</dna> ( <protein>PRIP</protein> ) ; Restriction Fragment Length Polymorphism ( <dna>RFLP</dna> ) analyses with four <dna>genomic anonymous probes</dna> ( <dna>D20S5</dna> , <dna>cD3H12</dna> , <dna>D20S17</dna> , <dna>D20S18</dna> ) ; and fluorescent in situ hybridization ( FISH ) with <dna>total human DNA</dna> and <dna>D20Z1</dna> , a sequence specific to the <dna>human Chr 20 centromere</dna> , as probes .


The NR2 hybrid allowed us to exclude three candidate genes for AGS : hepatic nuclear factor 3 beta ( HNF3 beta ) , paired box 1 ( PAX1 ) , and cystatin C ( CST3 ) as shown by their localization outside of the deletion .

The <cell_line>NR2 hybrid</cell_line> allowed us to exclude three <dna>candidate genes</dna> for AGS : <dna>hepatic nuclear factor 3 beta</dna> ( <dna>HNF3 beta</dna> ) , <dna>paired box 1</dna> ( <dna>PAX1</dna> ) , and <dna>cystatin C</dna> ( <dna>CST3</dna> ) as shown by their localization outside of the deletion .


The NR2 hybrid is a powerful tool for the mapping of new probes of this region , as well as for obtaining new informative probes specific for the deletion by subtractive cloning of the region .

The <cell_line>NR2 hybrid</cell_line> is a powerful tool for the mapping of new probes of this region , as well as for obtaining new informative probes specific for the deletion by subtractive cloning of the region .


Such markers will be useful for linkage analysis and screening of cDNA libraries .

Such markers will be useful for linkage analysis and screening of <dna>cDNA libraries</dna> .


Positive and negative regulation of the composite octamer motif of the interleukin 2 enhancer by AP-1 , Oct-2 , and retinoic acid receptor .

Positive and negative regulation of the <dna>composite octamer motif</dna> of the <dna>interleukin 2 enhancer</dna> by <protein>AP-1</protein> , <protein>Oct-2</protein> , and <protein>retinoic acid receptor</protein> .


The differentiating agent retinoic acid ( RA ) has been previously reported to interfere with 12-O-tetradecanoyl-phorbol-13-acetate ( TPA ) /Ca ( 2+ ) -induced signals for the regulation of the -96 to -66-bp octamer motif found in the enhancer for the interleukin ( IL ) -2 gene , which encodes a major T lymphocyte growth factor .

The differentiating agent retinoic acid ( RA ) has been previously reported to interfere with 12-O-tetradecanoyl-phorbol-13-acetate ( TPA ) /Ca ( 2+ ) -induced signals for the regulation of the <dna>-96 to -66-bp octamer motif</dna> found in the enhancer for the <dna>interleukin ( IL ) -2 gene</dna> , which encodes a major <protein>T lymphocyte growth factor</protein> .


The IL-2 octamer motif is a composite cis-element which binds Oct-1 and Oct-2 as well as a TPA/Ca ( 2+ ) -inducible nuclear factor , previously termed octamer-associated protein ( OAP40 ) .

The <dna>IL-2 octamer motif</dna> is a composite <dna>cis-element</dna> which binds <protein>Oct-1</protein> and <protein>Oct-2</protein> as well as a <protein>TPA/Ca ( 2+ ) -inducible nuclear factor</protein> , previously termed <protein>octamer-associated protein</protein> ( <protein>OAP40</protein> ) .


We show here that Oct-2 , despite the presence of an active transcriptional activation domain , requires TPA/Ca ( 2+ ) -induced signals to strongly transactivate the IL-2 octamer motif in Jurkat T cells .

We show here that <protein>Oct-2</protein> , despite the presence of an active transcriptional activation domain , requires TPA/Ca ( 2+ ) -induced signals to strongly transactivate the <dna>IL-2 octamer motif</dna> in Jurkat T cells .


This Oct-2 -dependent transactivation is inhibited by RA .

This <protein>Oct-2</protein> -dependent transactivation is inhibited by RA .


The presence of an intact COOH-terminal domain of Oct-2 contributes to both TPA/Ca ( 2+ ) -induced transactivation and the RA-mediated repression .

The presence of an <protein>intact COOH-terminal domain</protein> of <protein>Oct-2</protein> contributes to both TPA/Ca ( 2+ ) -induced transactivation and the RA-mediated repression .


We also show that both Fos and Jun components of the AP-1 factors participate in the OAP40 complex .

We also show that both <protein>Fos</protein> and <protein>Jun</protein> components of the <protein>AP-1</protein> factors participate in the <protein>OAP40 complex</protein> .


Furthermore , transfected c-jun , jun-B , jun-D , c-fos , or Fos-B expression vectors partially substitute for TPA and Ca2+ and cooperate with Oct-2 for the transactivation of the combined OAP/octamer cis-element .

Furthermore , transfected <dna>c-jun , jun-B , jun-D , c-fos , or Fos-B expression vectors</dna> partially substitute for TPA and Ca2+ and cooperate with <protein>Oct-2</protein> for the transactivation of the combined <dna>OAP/octamer cis-element</dna> .


Mutations of the genuine octamer-binding site abrogate both the binding of Oct-1 and Oct-2 and the TPA/Ca ( 2+ ) -induced transactivation of the OAP/octamer motif .

Mutations of the <dna>genuine octamer-binding site</dna> abrogate both the binding of <protein>Oct-1</protein> and <protein>Oct-2</protein> and the TPA/Ca ( 2+ ) -induced transactivation of the <dna>OAP/octamer motif</dna> .


OAP confers to Oct-2 responsivity to both TPA/Ca2+ and RA , since specific mutations of the AP-1/OAP-binding site significantly reduce the transactivation by Oct-2 in response to TPA and Ca2+ and abolish the inhibition by RA .

<protein>OAP</protein> confers to <protein>Oct-2</protein> responsivity to both TPA/Ca2+ and RA , since specific mutations of the <dna>AP-1/OAP-binding site</dna> significantly reduce the transactivation by <protein>Oct-2</protein> in response to TPA and Ca2+ and abolish the inhibition by RA .


Furthermore , retinoic acid receptor ( RAR ) alpha is able to inhibit in vitro the formation of the complex between the nuclear AP-1 /OAP and its specific binding site , resulting in the interference with Oct-2 -dependent cis-regulatory function of this AP-1 element .

Furthermore , <protein>retinoic acid receptor</protein> ( <protein>RAR</protein> ) alpha is able to inhibit in vitro the formation of the complex between the nuclear <protein>AP-1</protein> <protein>/OAP</protein> and its specific binding site , resulting in the interference with <protein>Oct-2</protein> -dependent cis-regulatory function of this <protein>AP-1</protein> element .


Therefore , we propose that the TPA/calcium-activated AP-1 /OAP element is the main target of positive or negative regulatory signals influencing the IL-2 octamer motif , through synergism with Oct-2 and antagonism by RAR .

Therefore , we propose that the TPA/calcium-activated <protein>AP-1</protein> <protein>/OAP</protein> element is the main target of positive or negative regulatory signals influencing the <dna>IL-2 octamer motif</dna> , through synergism with <protein>Oct-2</protein> and antagonism by <protein>RAR</protein> .


Missense mutation in exon 7 of the common gamma chain gene causes a moderate form of X-linked combined immunodeficiency .

Missense mutation in <dna>exon 7</dna> of the <dna>common gamma chain gene</dna> causes a moderate form of X-linked combined immunodeficiency .


Clinical and immunologic features of a recently recognized X-linked combined immunodeficiency disease ( XCID ) suggested that XCID and X-linked severe combined immunodeficiency ( XSCID ) might arise from different genetic defects .


The recent discovery of mutations in the common gamma chain ( gamma c ) gene , a constituent of several cytokine receptors , in XSCID provided an opportunity to test directly whether a previously unrecognized mutation in this same gene was responsible for XCID .

The recent discovery of mutations in the <dna>common gamma chain ( gamma c ) gene</dna> , a constituent of several <protein>cytokine receptors</protein> , in XSCID provided an opportunity to test directly whether a previously unrecognized mutation in this same gene was responsible for XCID .


The status of X chromosome inactivation in blood leukocytes from obligate carriers of XCID was determined from the polymorphic , short tandem repeats ( CAG ) , in the androgen receptor gene , which also contains a methylation-sensitive HpaII site .

The status of <dna>X chromosome</dna> inactivation in blood leukocytes from obligate carriers of XCID was determined from the polymorphic , short tandem repeats ( CAG ) , in the <dna>androgen receptor gene</dna> , which also contains a <dna>methylation-sensitive HpaII site</dna> .


As in XSCID , X-chromosome inactivation in obligate carriers of XCID was nonrandom in T and B lymphocytes .

As in XSCID , X-chromosome inactivation in obligate carriers of XCID was nonrandom in <cell_type>T and B lymphocytes</cell_type> .


In addition , X chromosome inactivation in PMNs was variable .

In addition , <dna>X chromosome</dna> inactivation in PMNs was variable .


Findings from this analysis prompted sequencing of the gamma c gene in this pedigree .

Findings from this analysis prompted sequencing of the <dna>gamma c gene</dna> in this pedigree .


A missense mutation in the region coding for the cytoplasmic portion of the gamma c gene was found in three affected males but not in a normal brother .

A missense mutation in the region coding for the cytoplasmic portion of the <dna>gamma c gene</dna> was found in three affected males but not in a normal brother .


Therefore , this point mutation in the gamma c gene leads to a less severe degree of deficiency in cellular and humoral immunity than that seen in XSCID .

Therefore , this point mutation in the <dna>gamma c gene</dna> leads to a less severe degree of deficiency in cellular and humoral immunity than that seen in XSCID .


Posttranscriptional regulation of macrophage tissue factor expression by antioxidants .

Posttranscriptional regulation of macrophage <protein>tissue factor</protein> expression by antioxidants .


Tissue factor ( TF ) expression by cells of monocyte/macrophage lineage represents an important mechanism underlying the initiation of fibrin deposition at sites of extravascular inflammation .

<protein>Tissue factor</protein> ( <protein>TF</protein> ) expression by cells of <cell_type>monocyte/macrophage lineage</cell_type> represents an important mechanism underlying the initiation of <protein>fibrin</protein> deposition at sites of extravascular inflammation .


Recent evidence suggests a role for oxidant stress in the signalling pathway of various cell types by virtue of its ability to induce DNA binding of various transcription factors , including nuclear factor kappa B and AP-1 .

Recent evidence suggests a role for oxidant stress in the signalling pathway of various cell types by virtue of its ability to induce DNA binding of various <protein>transcription factors</protein> , including <protein>nuclear factor kappa B</protein> and <protein>AP-1</protein> .


The effect of antioxidant treatment on lipopolysaccharide ( LPS ) -induced TF expression was examined in murine peritoneal macrophages and human monocytes .

The effect of antioxidant treatment on lipopolysaccharide ( LPS ) -induced <protein>TF</protein> expression was examined in <cell_type>murine peritoneal macrophages</cell_type> and <cell_type>human monocytes</cell_type> .


Both pyrrolidine dithiocarbamate , an oxidant scavenger , and N-acetyl-cysteine , a precursor of the endogenous antioxidant glutathione , inhibited stimulation of macrophage procoagulant activity by LPS .


Northern blot analysis showed that neither of these agents reduced LPS-stimulated TF mRNA accumulation , thereby suggesting a posttranscriptional mechanism for the effect .

Northern blot analysis showed that neither of these agents reduced LPS-stimulated <protein>TF</protein> mRNA accumulation , thereby suggesting a posttranscriptional mechanism for the effect .


Immunofluorescence studies of human monocytes using polyclonal anti-TF antibody showed that N-acetyl-cysteine treatment prevented the characteristic plasmalemmal localization of TF antigen that occurs in response to LPS .

Immunofluorescence studies of <cell_type>human monocytes</cell_type> using polyclonal <protein>anti-TF antibody</protein> showed that N-acetyl-cysteine treatment prevented the characteristic plasmalemmal localization of <protein>TF antigen</protein> that occurs in response to LPS .