PLB-985 cells represent a bipotential myelomonoblastic cell population capable of either granulocytic or monocytic differentiation after induction with appropriate inducers .

<cell_line>PLB-985 cells</cell_line> represent a <cell_line>bipotential myelomonoblastic cell population</cell_line> capable of either granulocytic or monocytic differentiation after induction with appropriate inducers .


By virtue of the presence of CD4 on the cell surface , PLB-985 cells were chronically infected with HIV-1 strain IIIB .

By virtue of the presence of <protein>CD4</protein> on the cell surface , <cell_line>PLB-985 cells</cell_line> were chronically infected with HIV-1 strain IIIB .


PLB-IIIB cells clearly possessed a more monocytic phenotype than the parental myeloblasts , as determined by differential staining , increased expression of the myeloid-specific surface markers , and transcription of the c-fms proto-oncogene .

<cell_line>PLB-IIIB cells</cell_line> clearly possessed a more monocytic phenotype than the <cell_type>parental myeloblasts</cell_type> , as determined by differential staining , increased expression of the <protein>myeloid-specific surface markers</protein> , and transcription of the <dna>c-fms proto-oncogene</dna> .


NF-kappa B binding activity was inducible by tumor necrosis factor and phorbol myristate acetate in PLB-985 .

<protein>NF-kappa B</protein> binding activity was inducible by <protein>tumor necrosis factor</protein> and phorbol myristate acetate in <cell_line>PLB-985</cell_line> .


However , in PLB-IIIB cells , constitutive expression of a novel NF-kappa B complex was detected , composed of proteins ranging between 70 and 110 kD .

However , in <cell_line>PLB-IIIB cells</cell_line> , constitutive expression of a novel <protein>NF-kappa B complex</protein> was detected , composed of proteins ranging between 70 and 110 kD .


These proteins interacted specifically with the symmetric NF-kappa B site from the interferon beta ( IFN-beta ) promoter .

These proteins interacted specifically with the <dna>symmetric NF-kappa B site</dna> from the <dna>interferon beta ( IFN-beta ) promoter</dna> .


Mutations affecting the 5 ' guanine residues of the kappa B site were unable to compete for these NF-kappa B-related proteins .

Mutations affecting the <dna>5 ' guanine residues</dna> of the <dna>kappa B site</dna> were unable to compete for these <protein>NF-kappa B-related proteins</protein> .


Inducibility of endogenous IFN-beta and IFN-alpha RNA was also increased in PLB-IIIB cells .

Inducibility of endogenous <rna>IFN-beta and IFN-alpha RNA</rna> was also increased in <cell_line>PLB-IIIB cells</cell_line> .


These studies indicate that HIV-1 infection of myelomonoblastic cells may select for a more mature monocytic phenotype and that unique subunit associations of NF-kappa B DNA binding proteins may contribute to differential NF-kappa B -mediated gene expression .

These studies indicate that HIV-1 infection of <cell_type>myelomonoblastic cells</cell_type> may select for a more mature monocytic phenotype and that unique subunit associations of <protein>NF-kappa B DNA binding proteins</protein> may contribute to differential <protein>NF-kappa B</protein> -mediated gene expression .


Cortisol receptor resistance : the variability of its clinical presentation and response to treatment .


Primary ( partial ) cortisol receptor resistance was previously reported in a total of 7 patients and 14 asymptomatic family members .

Primary ( partial ) <protein>cortisol receptor</protein> resistance was previously reported in a total of 7 patients and 14 asymptomatic family members .


Its occurrence is considered to be extremely rare .


In the present study we report on 6 patients ( 2 males and 4 females ) with the syndrome .


The first male patient presented with mild hypertension .


Hydrochlorothiazide therapy resulted in life-threatening hypokalemia .


The second male patient had slight hypertension without hypokalemia .


All four female patients presented between the age of 20-30 yr with acne , hirsutism , and irregular menstruations .


Low dose dexamethasone therapy ( 1-1.5 mg/day ) was of clinical benefit in these patients .


All patients showed insufficient suppression of serum cortisol concentrations in the overnight 1-mg dexamethasone test .


The diurnal rhythm of ACTH and cortisol was intact , albeit at an elevated level .


There was a normal increase in ACTH , cortisol , and GH ( except in one obese patient ) in response to insulin-induced hypoglycemia , while cortisol production was elevated in three patients .


Circulating adrenal androgen levels were increased in all patients .


Glucocorticoid receptors were investigated in a whole cell dexamethasone binding assay in mononuclear leukocytes .

<protein>Glucocorticoid receptors</protein> were investigated in a whole cell dexamethasone binding assay in <cell_type>mononuclear leukocytes</cell_type> .


In the first male patient , the number of receptors was very low , while the affinity was lower than that in controls .


A lowered affinity to dexamethasone was found in one female patient , while a lowered number of receptors was found in three patients .


In the second male patient , no abnormalities were found .


As a bioassay for glucocorticoid action we also measured dexamethasone suppressibility of mitogen-stimulated incorporation of [ 3H ] thymidine in mononuclear leukocytes .

As a bioassay for glucocorticoid action we also measured dexamethasone suppressibility of mitogen-stimulated incorporation of [ 3H ] thymidine in <cell_type>mononuclear leukocytes</cell_type> .


In the male patient with normal receptor status , dexamethasone suppressibility of [ 3H ] thymidine incorporation was significantly lower than that in healthy controls with respect to both maximal suppression and IC50 .


Partial cortisol receptor resistance might be less rare than previously thought .


In the six patients presented , at least three different forms can be recognized .


Therapy with dexamethasone was successful in female patients with acne and hirsutism , as the secondary increase in the production of adrenal androgens was effectively controlled .


The development of functionally responsive T cells .

The development of functionally responsive <cell_type>T cells</cell_type> .


The work reviewed in this article separates T cell development into four phases .


First is an expansion phase prior to TCR rearrangement , which appears to be correlated with programming of at least some response genes for inducibility .

First is an expansion phase prior to <protein>TCR</protein> rearrangement , which appears to be correlated with programming of at least some response genes for inducibility .


This phase can occur to some extent outside of the thymus .


However , the profound T cell deficit of nude mice indicates that the thymus is by far the most potent site for inducing the expansion per se , even if other sites can induce some response acquisition .


Second is a controlled phase of TCR gene rearrangement .

Second is a controlled phase of <dna>TCR gene</dna> rearrangement .


The details of the regulatory mechanism that selects particular loci for rearrangement are still not known .

The details of the regulatory mechanism that selects particular <dna>loci</dna> for rearrangement are still not known .


It seems that the rearrangement of the TCR gamma loci in the gamma delta lineage may not always take place at a developmental stage strictly equivalent to the rearrangement of TCR beta in the alpha beta lineage , and it is not clear just how early the two lineages diverge .

It seems that the rearrangement of the <dna>TCR gamma loci</dna> in the <cell_type>gamma delta lineage</cell_type> may not always take place at a developmental stage strictly equivalent to the rearrangement of <dna>TCR beta</dna> in the <cell_type>alpha beta lineage</cell_type> , and it is not clear just how early the two lineages diverge .


In the TCR alpha beta lineage , however , the final gene rearrangement events are accompanied by rapid proliferation and an interruption in cellular response gene inducibility .

In the <cell_type>TCR alpha beta lineage ,</cell_type> however , the final gene rearrangement events are accompanied by rapid proliferation and an interruption in cellular response gene inducibility .


The loss of conventional responsiveness is probably caused by alterations at the level of signaling , and may be a manifestation of the physiological state that is a precondition for selection .


Third is the complex process of selection .


Whereas peripheral T cells can undergo forms of positive selection ( by antigen-driven clonal expansion ) and negative selection ( by abortive stimulation leading to anergy or death ) , neither is exactly the same phenomenon that occurs in the thymic cortex .

Whereas <cell_type>peripheral T cells</cell_type> can undergo forms of positive selection ( by antigen-driven clonal expansion ) and negative selection ( by abortive stimulation leading to anergy or death ) , neither is exactly the same phenomenon that occurs in the thymic cortex .


Negative selection in the cortex appears to be a suicidal inversion of antigen responsiveness : instead of turning on IL-2 expression , the activated cell destroys its own chromatin .

Negative selection in the cortex appears to be a suicidal inversion of antigen responsiveness : instead of turning on <protein>IL-2</protein> expression , the <cell_type>activated cell</cell_type> destroys its own <dna>chromatin</dna> .


The genes that need to be induced for this response are not yet identified , but it is unquestionably a form of activation .


It is interesting that in humans and rats , cortical thymocytes undergoing negative selection can still induce IL-2R alpha expression and even be rescued in vitro , if exogenous IL-2 is provided .

It is interesting that in humans and rats , <cell_type>cortical thymocytes</cell_type> undergoing negative selection can still induce <protein>IL-2R alpha</protein> expression and even be rescued in vitro , if <protein>exogenous IL-2</protein> is provided .


Perhaps murine thymocytes are denied this form of rescue because they shut off IL-2R beta chain expression at an earlier stage or because they may be uncommonly Bcl-2 deficient ( cf. Sentman et al. , 1991 ; Strasser et al. , 1991 ) .

Perhaps <cell_type>murine thymocytes</cell_type> are denied this form of rescue because they shut off <protein>IL-2R beta chain</protein> expression at an earlier stage or because they may be uncommonly <protein>Bcl-2</protein> deficient ( cf. Sentman et al. , 1991 ; Strasser et al. , 1991 ) .


Even so , medullary thymocytes remain at least partially susceptible to negative selection even as they continue to mature .

Even so , <cell_type>medullary thymocytes</cell_type> remain at least partially susceptible to negative selection even as they continue to mature .


I kappa B/MAD-3 masks the nuclear localization signal of NF-kappa B p65 and requires the transactivation domain to inhibit NF-kappa B p65 DNA binding .

<protein>I kappa B/MAD-3</protein> masks the nuclear localization signal of <protein>NF-kappa B p65</protein> and requires the <protein>transactivation domain</protein> to inhibit <protein>NF-kappa B p65</protein> DNA binding .


The active nuclear form of the NF-kappa B transcription factor complex is composed of two DNA binding subunits , NF-kappa B p65 and NF-kappa B p50 , both of which share extensive N-terminal sequence homology with the v-rel oncogene product .

The active nuclear form of the <protein>NF-kappa B transcription factor complex</protein> is composed of two <protein>DNA binding subunits</protein> , <protein>NF-kappa B p65</protein> and <protein>NF-kappa B p50</protein> , both of which share extensive N-terminal sequence homology with the <protein>v-rel oncogene product</protein> .


The NF-kappa B p65 subunit provides the transactivation activity in this complex and serves as an intracellular receptor for a cytoplasmic inhibitor of NF-kappa B , termed I kappa B .

The <protein>NF-kappa B p65 subunit</protein> provides the transactivation activity in this complex and serves as an intracellular receptor for a cytoplasmic inhibitor of <protein>NF-kappa B</protein> , termed I kappa B .


In contrast , NF-kappa B p50 alone fails to stimulate kappa B-directed transcription , and based on prior in vitro studies , is not directly regulated by I kappa B .

In contrast , <protein>NF-kappa B p50</protein> alone fails to stimulate kappa B-directed transcription , and based on prior in vitro studies , is not directly regulated by <protein>I kappa B</protein> .


To investigate the molecular basis for the critical regulatory interaction between NF-kappa B and I kappa B/MAD-3 , a series of human NF-kappa B p65 mutants was identified that functionally segregated DNA binding , I kappa B -mediated inhibition , and I kappa B -induced nuclear exclusion of this transcription factor .

To investigate the molecular basis for the critical regulatory interaction between <protein>NF-kappa B</protein> and <protein>I kappa B/MAD-3</protein> , a series of <protein>human NF-kappa B p65 mutants</protein> was identified that functionally segregated DNA binding , <protein>I kappa B</protein> -mediated inhibition , and <protein>I kappa B</protein> -induced nuclear exclusion of this <protein>transcription factor</protein> .


Results from in vivo expression studies performed with these NF-kappa B p65 mutants revealed the following : 1 ) I kappa B/MAD-3 completely inhibits NF-kappa B p65 -dependent transcriptional activation mediated through the human immunodeficiency virus type 1 kappa B enhancer in human T lymphocytes , 2 ) the binding of I kappa B/MAD-3 to NF-kappa B p65 is sufficient to retarget NF-kappa B p65 from the nucleus to the cytoplasm , 3 ) selective deletion of the functional nuclear localization signal present in the Rel homology domain of NF-kappa B p65 disrupts its ability to engage I kappa B/MAD-3 , and 4 ) the unique C-terminus of NF-kappa B p65 attenuates its own nuclear localization and contains sequences that are required for I kappa B -mediated inhibition of NF-kappa B p65 DNA binding activity .

Results from in vivo expression studies performed with these <protein>NF-kappa B p65 mutants</protein> revealed the following : 1 ) <protein>I kappa B/MAD-3</protein> completely inhibits <protein>NF-kappa B p65</protein> -dependent transcriptional activation mediated through the <dna>human immunodeficiency virus type 1 kappa B enhancer</dna> in <cell_type>human T lymphocytes</cell_type> , 2 ) the binding of <protein>I kappa B/MAD-3</protein> to <protein>NF-kappa B p65</protein> is sufficient to retarget <protein>NF-kappa B p65</protein> from the nucleus to the cytoplasm , 3 ) selective deletion of the functional nuclear localization signal present in the <protein>Rel homology domain</protein> of <protein>NF-kappa B p65</protein> disrupts its ability to engage <protein>I kappa B/MAD-3</protein> , and 4 ) the unique <protein>C-terminus</protein> of <protein>NF-kappa B p65</protein> attenuates its own nuclear localization and contains sequences that are required for <protein>I kappa B</protein> -mediated inhibition of <protein>NF-kappa B p65</protein> DNA binding activity .


Together , these findings suggest that the nuclear localization signal and transactivation domain of NF-kappa B p65 constitute a bipartite system that is critically involved in the inhibitory function of I kappa B/MAD-3 .

Together , these findings suggest that the nuclear localization signal and <protein>transactivation domain</protein> of <protein>NF-kappa B p65</protein> constitute a bipartite system that is critically involved in the inhibitory function of <protein>I kappa B/MAD-3</protein> .


Unexpectedly , our in vivo studies also demonstrate that I kappa B/MAD-3 binds directly to NF-kappa B p50 .

Unexpectedly , our in vivo studies also demonstrate that <protein>I kappa B/MAD-3</protein> binds directly to <protein>NF-kappa B p50</protein> .


This interaction is functional as it leads to retargeting of NF-kappa B p50 from the nucleus to the cytoplasm .

This interaction is functional as it leads to retargeting of <protein>NF-kappa B p50</protein> from the nucleus to the cytoplasm .


However , no loss of DNA binding activity is observed , presumably reflecting the unique C-terminal domain that is distinct from that present in NF-kappa B p65 .

However , no loss of DNA binding activity is observed , presumably reflecting the unique <protein>C-terminal domain</protein> that is distinct from that present in <protein>NF-kappa B p65</protein> .


Characterization of a new tissue-specific transcription factor binding to the simian virus 40 enhancer TC-II ( NF-kappa B ) element .

Characterization of a new tissue-specific <protein>transcription factor</protein> binding to the <dna>simian virus 40 enhancer TC-II ( NF-kappa B ) element</dna> .


We have biochemically and functionally characterized a new transcription factor , NP-TCII , which is present in nuclei from unstimulated T and B lymphocytes but is not found in nonhematopoietic cells .

We have biochemically and functionally characterized a new <protein>transcription factor</protein> , <protein>NP-TCII</protein> , which is present in nuclei from <cell_line>unstimulated T and B lymphocytes</cell_line> but is not found in <cell_type>nonhematopoietic cells</cell_type> .


This factor has a DNA-binding specificity similar to that of NF-kappa B but is unrelated to this or other Rel proteins by functional and biochemical criteria .

This factor has a DNA-binding specificity similar to that of <protein>NF-kappa B</protein> but is unrelated to this or other <protein>Rel proteins</protein> by functional and biochemical criteria .


It can also be distinguished from other previously described lymphocyte-specific DNA-binding proteins .

It can also be distinguished from other previously described <protein>lymphocyte-specific DNA-binding proteins</protein> .


The candidate oncoprotein Bcl-3 is an antagonist of p50/NF-kappa B -mediated inhibition .

The <protein>candidate oncoprotein Bcl-3</protein> is an antagonist of <protein>p50/NF-kappa B</protein> -mediated inhibition .


The candidate oncogene bcl-3 was discovered as a translocation into the immunoglobulin alpha-locus in some cases of B-cell chronic lymphocytic leukaemias .

The <dna>candidate oncogene bcl-3</dna> was discovered as a translocation into the <dna>immunoglobulin alpha-locus</dna> in some cases of B-cell chronic lymphocytic leukaemias .


The protein Bcl-3 contains seven so-called ankyrin repeats .

The <protein>protein Bcl-3</protein> contains seven so-called <protein>ankyrin repeats</protein> .


Similar repeat motifs are found in a number of diverse regulatory proteins but the motifs of Bcl-3 are most closely related to those found in I kappa B proteins in which the ankyrin repeat domain is thought to be directly involved in inhibition of NF-kappa B activity .

Similar <protein>repeat motifs</protein> are found in a number of <protein>diverse regulatory proteins</protein> but the motifs of <protein>Bcl-3</protein> are most closely related to those found in <protein>I kappa B proteins</protein> in which the <protein>ankyrin repeat</protein> domain is thought to be directly involved in inhibition of <protein>NF-kappa B</protein> activity .


No biological function has yet been described for Bcl-3 , but it was noted recently that Bcl-3 interferes with DNA-binding of the p50 subunit of NF-kappa B in vitro .

No biological function has yet been described for <protein>Bcl-3</protein> , but it was noted recently that <protein>Bcl-3</protein> interferes with DNA-binding of the <protein>p50 subunit</protein> of <protein>NF-kappa B</protein> in vitro .


Here we demonstrate that Bcl-3 can aid kappa B site -dependent transcription in vivo by counteracting the inhibitory effects of p50/NF-kappa B homodimers .

Here we demonstrate that <protein>Bcl-3</protein> can aid <dna>kappa B site</dna> -dependent transcription in vivo by counteracting the inhibitory effects of <protein>p50/NF-kappa B homodimers</protein> .


Bcl-3 may therefore aid activation of select NF-kappa B-regulated genes , including those of the human immunodeficiency virus .

<protein>Bcl-3</protein> may therefore aid activation of select <dna>NF-kappa B-regulated genes</dna> , including those of the human immunodeficiency virus .


[ Mechanism of action of steroid hormones. I. Estrogens ]


The steroid hormone are very versatile molecules : although they are related among them by their chemical structure , they have very diverse functions and including antagonic .


Their action mechanism is not completely cleared .


The estrogens participate in the regulation of practically all the reproductive and sexual events of the female , although the intracellular actions by which they take place are not well known and the proposed models do not adequately satisfy the questions .


Currently it is accepted the existence of a cytoplasmic and/or nuclear receptor , without explaining satisfactorily how the hormones come to the nucleus .

Currently it is accepted the existence of a <protein>cytoplasmic and/or nuclear receptor</protein> , without explaining satisfactorily how the hormones come to the nucleus .


The endocrine events that are rapidly expressed ( seconds ) are due to a possible interaction with cellular membrane .


The purpose of this review is to analyze and concilliate the reported data on the mechanism of action of estrogens .


Reduced susceptibility to HIV-1 infection of ethyl-methanesulfonate-treated CEM subclones correlates with a blockade in their protein kinase C signaling pathway .

Reduced susceptibility to HIV-1 infection of <cell_line>ethyl-methanesulfonate-treated CEM subclones</cell_line> correlates with a blockade in their <protein>protein kinase C</protein> signaling pathway .


We have described the isolation of chemically induced CEM subclones that express CD4 receptors and bind soluble gp120 , yet show a markedly reduced susceptibility to infection with HIV-1 .

We have described the isolation of chemically induced <cell_line>CEM subclones</cell_line> that express <protein>CD4 receptors</protein> and bind soluble <protein>gp120</protein> , yet show a markedly reduced susceptibility to infection with HIV-1 .


Two subclones were found to have an abnormal response to the protein kinase C ( PKC ) activator PMA .

Two subclones were found to have an abnormal response to the <protein>protein kinase C</protein> ( <protein>PKC</protein> ) activator PMA .


PMA treatment induced CD3 and CD25 ( IL-2R ) receptors on the parental line and on other ethyl-methanesulfonate-derived subclones , but not on these two mutants .

PMA treatment induced <protein>CD3 and CD25 ( IL-2R ) receptors</protein> on the parental line and on other <cell_line>ethyl-methanesulfonate-derived subclones</cell_line> , but not on these two mutants .


Direct assays of PKC activity were conducted .

Direct assays of <protein>PKC</protein> activity were conducted .


Total cellular PKC enzymatic activity was found to be normal in these subclones .

Total cellular <protein>PKC</protein> enzymatic activity was found to be normal in these subclones .


PMA-induced CD4 down-modulation occurred normally .

PMA-induced <protein>CD4</protein> down-modulation occurred normally .


In addition , activation of c-raf kinase was normal .

In addition , activation of <protein>c-raf kinase</protein> was normal .


Since HIV-1 long terminal repeat contains two functional nuclear factor kB ( NF-kB ) regulatory elements , we studied the ability of PMA to induce NF-kB binding activity by different assays .

Since <dna>HIV-1 long terminal repeat</dna> contains two functional <dna>nuclear factor kB ( NF-kB ) regulatory elements</dna> , we studied the ability of PMA to induce <protein>NF-kB</protein> binding activity by different assays .


Chloramphenicol acetyl transferase ( CAT ) assays using the HIV-1 ( -139 ) long terminal repeat-CAT construct showed no PMA induction of CAT activity in these subclones ( unlike the parental line and other subclones ) .

<protein>Chloramphenicol acetyl transferase</protein> ( <protein>CAT</protein> ) assays using the <dna>HIV-1 ( -139 ) long terminal repeat-CAT construct</dna> showed no PMA induction of <protein>CAT</protein> activity in these subclones ( unlike the parental line and other subclones ) .


Okadaic acid , an inhibitor of phosphatases 1 and 2A , did not overcome the defect in these subclones .

Okadaic acid , an inhibitor of <protein>phosphatases 1 and 2A</protein> , did not overcome the defect in these subclones .


Gel retardation assays , using a 32P-probe containing the HIV-1 NF-kB probe and nuclear extracts from PMA-treated cells , showed significantly reduced induction of nuclear NF-kB binding proteins in these two subclones compared with wild type CEM and a control subclone .

Gel retardation assays , using a 32P-probe containing the HIV-1 <protein>NF-kB</protein> probe and nuclear extracts from <cell_type>PMA-treated cells</cell_type> , showed significantly reduced induction of <protein>nuclear NF-kB binding proteins</protein> in these two subclones compared with <protein>wild type CEM</protein> and a control subclone .


Deoxycholate treatment of cytoplasmic extracts from these subclones released much reduced NF-kB binding proteins from their cytoplasmic pools .

Deoxycholate treatment of cytoplasmic extracts from these subclones released much reduced <protein>NF-kB binding proteins</protein> from their cytoplasmic pools .


Thus , reduced levels of PKC -induced nuclear NF-kB activity in two T cell subclones did not affect their normal cell growth , but correlated with a pronounced reduction in their susceptibility to HIV-1 infection .

Thus , reduced levels of <protein>PKC</protein> -induced nuclear <protein>NF-kB</protein> activity in two T cell subclones did not affect their normal cell growth , but correlated with a pronounced reduction in their susceptibility to HIV-1 infection .


Nuclear factor of activated T cells contains Fos and Jun .

<protein>Nuclear factor</protein> of <cell_type>activated T cells</cell_type> contains <protein>Fos</protein> and <protein>Jun</protein> .


The nuclear factor NF-AT ( ref. 1 ) is induced in T cells stimulated through the T-cell receptor/CD3 complex , and is required for interleukin-2 ( IL-2 ) gene induction .

The <protein>nuclear factor NF-AT</protein> ( ref. 1 ) is induced in <cell_type>T cells</cell_type> stimulated through the <protein>T-cell receptor/CD3 complex</protein> , and is required for <dna>interleukin-2 ( IL-2 ) gene</dna> induction .


Although NF-AT has not been cloned or purified , there is evidence that it is a major target for immunosuppression by cyclosporin A ( CsA ) and FK506 ( refs 2-7 ) .

Although <protein>NF-AT</protein> has not been cloned or purified , there is evidence that it is a major target for immunosuppression by cyclosporin A ( CsA ) and FK506 ( refs 2-7 ) .


NF-AT induction may require two activation-dependent events : the CsA-sensitive translocation of a pre-existing component and the CsA-resistant synthesis of a nuclear component .

<protein>NF-AT</protein> induction may require two activation-dependent events : the CsA-sensitive translocation of a pre-existing component and the CsA-resistant synthesis of a <protein>nuclear component</protein> .


Here we report that the newly synthesized nuclear component of NF-AT is the transcription factor AP-1 .

Here we report that the newly synthesized <protein>nuclear component</protein> of <protein>NF-AT</protein> is the <protein>transcription factor</protein> <protein>AP-1</protein> .


We show that the inducible nuclear form of NF-AT contains Fos and Jun proteins .

We show that the inducible nuclear form of <protein>NF-AT</protein> contains <protein>Fos</protein> and <protein>Jun</protein> proteins .


Furthermore , we identify a pre-existing NF-AT-binding factor that is present in hypotonic extracts of unstimulated T cells .

Furthermore , we identify a <protein>pre-existing NF-AT-binding factor</protein> that is present in hypotonic extracts of <cell_type>unstimulated T cells</cell_type> .


On the basis of binding , reconstitution and cotransfection experiments , we propose that activation of NF-AT occurs in at least two stages : a CsA-sensitive stage involving modification and/or translocation of the pre-existing NF-AT complex , and a CsA-insensitive stage involving the addition of newly synthesized Fos or Fos/Jun proteins to the pre-existing complex .

On the basis of binding , reconstitution and cotransfection experiments , we propose that activation of <protein>NF-AT</protein> occurs in at least two stages : a CsA-sensitive stage involving modification and/or translocation of the pre-existing <protein>NF-AT complex</protein> , and a CsA-insensitive stage involving the addition of newly synthesized <protein>Fos</protein> or <protein>Fos/Jun proteins</protein> to the <protein>pre-existing complex</protein> .


A lymphoid cell-specific nuclear factor containing c-Rel-like proteins preferentially interacts with interleukin-6 kappa B-related motifs whose activities are repressed in lymphoid cells .

A <protein>lymphoid cell-specific nuclear factor</protein> containing <protein>c-Rel-like proteins</protein> preferentially interacts with <dna>interleukin-6 kappa B-related motifs</dna> whose activities are repressed in <cell_type>lymphoid cells</cell_type> .


The proto-oncoprotein c-Rel is a member of the nuclear factor kappa B transcription factor family , which includes the p50 and p65 subunits of nuclear factor kappa B .

The <protein>proto-oncoprotein c-Rel</protein> is a member of the <protein>nuclear factor kappa B transcription factor family</protein> , which includes the <protein>p50</protein> and <protein>p65 subunits</protein> of <protein>nuclear factor kappa B</protein> .


We show here that c-Rel binds to kappa B sites as homodimers as well as heterodimers with p50 .

We show here that <protein>c-Rel</protein> binds to <dna>kappa B sites</dna> as <protein>homodimers</protein> as well as <protein>heterodimers</protein> with <protein>p50</protein> .


These homodimers and heterodimers show distinct DNA-binding specificities and affinities for various kappa B motifs .

These <protein>homodimers</protein> and <protein>heterodimers</protein> show distinct DNA-binding specificities and affinities for various <dna>kappa B motifs</dna> .


In particular , the c-Rel homodimer has a high affinity for interleukin-6 ( IL-6 ) and beta interferon kappa B sites .

In particular , the <protein>c-Rel homodimer</protein> has a high affinity for <protein>interleukin-6</protein> ( <protein>IL-6</protein> ) and <dna>beta interferon kappa B sites</dna> .


In spite of its association with p50 in vitro , however , we found a lymphoid cell-specific nuclear factor in vivo that contains c-Rel but not p50 epitopes ; this factor , termed IL-6 kappa B binding factor II , appears to contain the c-Rel homodimer and preferentially recognizes several IL-6 kappa B-related kappa B motifs .

In spite of its association with <protein>p50</protein> in vitro , however , we found a <protein>lymphoid cell-specific nuclear factor</protein> in vivo that contains <protein>c-Rel</protein> but not <protein>p50 epitopes</protein> ; this factor , termed <protein>IL-6 kappa B binding factor II</protein> , appears to contain the <protein>c-Rel homodimer</protein> and preferentially recognizes several <dna>IL-6 kappa B-related kappa B motifs</dna> .


Although it has been previously shown that the IL-6 kappa B motif functions as a potent IL-1/tumor necrosis factor-responsive element in nonlymphoid cells , its activity was found to be repressed in lymphoid cells such as a Jurkat T-cell line .

Although it has been previously shown that the <dna>IL-6 kappa B motif</dna> functions as a potent <dna>IL-1/tumor necrosis factor-responsive element</dna> in <cell_type>nonlymphoid cells</cell_type> , its activity was found to be repressed in <cell_type>lymphoid cells</cell_type> such as a <cell_line>Jurkat T-cell line</cell_line> .


We also present evidence that IL-6 kappa B binding factor II functions as a repressor specific for IL-6 kappa B-related kappa B motifs in lymphoid cells .

We also present evidence that <protein>IL-6 kappa B binding factor II</protein> functions as a repressor specific for <dna>IL-6 kappa B-related kappa B motifs</dna> in <cell_type>lymphoid cells</cell_type> .


Cortisol resistance in acquired immunodeficiency syndrome .


This study concerns 9 iv drug abusers with acquired immunodeficiency syndrome ( AIDS ) who developed hypercortisolism without the clinical signs or metabolic consequences of hypercortisolism .


All patients were characterized by an Addisonian picture ( weakness , weight loss , hypotension , hyponatremia , and intense mucocutaneous melanosis ) .


An acquired form of peripheral resistance to glucocorticoids was suspected .


We , therefore , examined glucocorticoid receptor characteristics on mononuclear leukocytes by measuring [ 3H ] dexamethasone binding and the effect of dexamethasone on [ 3H ] thymidine incorporation , which is one of the effects of glucocorticoid receptor activation .

We , therefore , examined <protein>glucocorticoid receptor</protein> characteristics on <cell_type>mononuclear leukocytes</cell_type> by measuring [ 3H ] dexamethasone binding and the effect of dexamethasone on [ 3H ] thymidine incorporation , which is one of the effects of <protein>glucocorticoid receptor</protein> activation .


Glucocorticoid receptor density was increased in AIDS patients with an Addisonian picture ( group 1 ; 16.2 +/- 9.4 fmol/million cells ) compared to values in 12 AIDS patients without an Addisonian picture ( group 2 ; 6.05 +/- 2.6 fmol/million cells ; P less than 0.01 ) and sex- and age-matched controls ( 3.15 +/- 2.3 fmol/million cells ; P less than 0.01 ) .

<protein>Glucocorticoid receptor</protein> density was increased in AIDS patients with an Addisonian picture ( group 1 ; 16.2 +/- 9.4 fmol/million cells ) compared to values in 12 AIDS patients without an Addisonian picture ( group 2 ; 6.05 +/- 2.6 fmol/million cells ; P less than 0.01 ) and sex- and age-matched controls ( 3.15 +/- 2.3 fmol/million cells ; P less than 0.01 ) .


The affinity of glucocorticoid receptors ( Kd ) was strikingly decreased ( 9.36 +/- 3.44 nM in group 1 ; 3.2 +/- 1.5 nM in group 2 ; 2.0 +/- 0.8 nM in controls ; P less than 0.01 ) .

The affinity of <protein>glucocorticoid receptors</protein> ( Kd ) was strikingly decreased ( 9.36 +/- 3.44 nM in group 1 ; 3.2 +/- 1.5 nM in group 2 ; 2.0 +/- 0.8 nM in controls ; P less than 0.01 ) .


[ 3H ] Thymidine incorporation was decreased dose-dependently by dexamethasone in controls and patients ; the effect was significantly blunted ( P less than 0.05 ) in group 1 patients , which suggests that activation of glucocorticoid receptor is impaired as a result of the glucocorticoid receptor abnormality .

[ 3H ] Thymidine incorporation was decreased dose-dependently by dexamethasone in controls and patients ; the effect was significantly blunted ( P less than 0.05 ) in group 1 patients , which suggests that activation of <protein>glucocorticoid receptor</protein> is impaired as a result of the <protein>glucocorticoid receptor</protein> abnormality .


In conclusion , AIDS patients with hypercortisolism and clinical features of peripheral resistance to glucocorticoids are characterized by abnormal glucocorticoid receptors on lymphocytes .

In conclusion , AIDS patients with hypercortisolism and clinical features of peripheral resistance to glucocorticoids are characterized by <protein>abnormal glucocorticoid receptors</protein> on <cell_type>lymphocytes</cell_type> .


Resistance to glucocorticoids implies a complex change in immune-endocrine function , which may be important in the course of immunodeficiency syndrome .


A novel primer extension method to detect the number of CAG repeats in the androgen receptor gene in families with X-linked spinal and bulbar muscular atrophy .

A novel primer extension method to detect the number of CAG repeats in the <dna>androgen receptor gene</dna> in families with X-linked spinal and bulbar muscular atrophy .


X-linked spinal and bulbar muscular atrophy ( SBMA ) , an adult-onset form of motor neuron disease , was recently reported to be caused by amplification of the CAG repeats in the androgen receptor gene .

X-linked spinal and bulbar muscular atrophy ( SBMA ) , an adult-onset form of motor neuron disease , was recently reported to be caused by amplification of the CAG repeats in the <dna>androgen receptor gene</dna> .


We report here a simple and rapid strategy to detect the precise number of the CAGs .


After the DNA fragment containing the CAG repeats is amplified by the polymerase chain reaction , a primer extension is carried out ; the extension of the end-labelled reverse primer adjacent to 3 ' end of CAG repeats stops at the first T after CAG repeats with the incorporation of dideoxy ATP in the reaction mixture .

After the <dna>DNA fragment</dna> containing the <dna>CAG repeats</dna> is amplified by the polymerase chain reaction , a primer extension is carried out ; the extension of the <dna>end-labelled reverse primer</dna> adjacent to <dna>3 ' end</dna> of <dna>CAG repeats</dna> stops at the first T after <dna>CAG repeats</dna> with the incorporation of dideoxy ATP in the reaction mixture .


The resultant primer products are analysed by denaturing polyacrylamide gel electrophoresis and autoradiography .

The resultant <dna>primer products</dna> are analysed by denaturing polyacrylamide gel electrophoresis and autoradiography .


This method could be quite useful to detect not only CAG repeats in SBMA but also other polymorphic dinucleotide and trinucleotide repeats .

This method could be quite useful to detect not only <dna>CAG repeats</dna> in SBMA but also other <dna>polymorphic dinucleotide</dna> and <dna>trinucleotide repeats</dna> .


Cellular immune and cytokine pathways resulting in tissue factor expression and relevance to septic shock .

Cellular immune and cytokine pathways resulting in <protein>tissue factor</protein> expression and relevance to septic shock .


Cells of monocyte lineage serve as effector cells in the cellular immune response .

Cells of monocyte lineage serve as <cell_type>effector cells</cell_type> in the cellular immune response .


In addition , they respond to LPS and cytokines with activation and expression of inflammatory effector gene products similar to those elicited by the antigen driven response .

In addition , they respond to LPS and cytokines with activation and expression of <protein>inflammatory effector gene products</protein> similar to those elicited by the antigen driven response .


The response to antigen proceeds at the T helper cell level through two independent forms of cellular collaboration , contact and lymphokine .

The response to antigen proceeds at the <cell_type>T helper cell</cell_type> level through two independent forms of cellular collaboration , contact and <protein>lymphokine</protein> .


We review the control of expression of the Tissue Factor ( TF ) gene and the function of the TF protein .

We review the control of expression of the <dna>Tissue Factor ( TF ) gene</dna> and the function of the <protein>TF protein</protein> .


The enhanced initiation of transcription of the TF gene appears to require engagement of a 56 bp LPS Response Element , an enhancer that is engaged by both AP-1 type heterodimeric complexes as well as NF kappa B like heterodimeric complexes .

The enhanced initiation of transcription of the TF gene appears to require engagement of a 56 bp <dna>LPS Response Element</dna> , an enhancer that is engaged by both <protein>AP-1 type heterodimeric complexes</protein> as well as <protein>NF kappa B like heterodimeric complexes</protein> .


Dissociation of NF kappa B from Ig kappa B by cytokine and LPS stimulation , and possibly activated T cells , may represent a common pathway to induction of the TF and other inflammatory genes .

Dissociation of <protein>NF kappa B</protein> from <protein>Ig kappa B</protein> by cytokine and LPS stimulation , and possibly <cell_type>activated T cells</cell_type> , may represent a common pathway to induction of the <dna>TF and other inflammatory genes</dna> .


Enhancement of expression of TF is observed upon adhesion of Mo to endothelial cells and extracellular matrix proteins , as well as upon engagement of leukocyte integrins .

Enhancement of expression of <protein>TF</protein> is observed upon adhesion of Mo to <cell_type>endothelial cells</cell_type> and extracellular matrix proteins , as well as upon engagement of <protein>leukocyte integrins</protein> .


The biological effects that follow from expression of TF by vascular cells have been resolved by analysis of function aided by the use of recombinant full length TF and truncated surface domain of TF .

The biological effects that follow from expression of <protein>TF</protein> by vascular cells have been resolved by analysis of function aided by the use of <protein>recombinant full length TF</protein> and <protein>truncated surface domain</protein> of <protein>TF</protein> .


The rules of assembly of the cognate ligands of TF , namely the zymogen plasma factors VII and the serine protease factor VIIa , with the soluble surface domain of TF in free solution , in the presence of phospholipid surfaces and cell surface and of the anchored TF molecule have been described .

The rules of assembly of the cognate ligands of <protein>TF</protein> , namely the <protein>zymogen plasma factors VII</protein> and the <protein>serine protease factor VIIa</protein> , with the <protein>soluble surface domain</protein> of <protein>TF</protein> in free solution , in the presence of phospholipid surfaces and cell surface and of the <protein>anchored TF molecule</protein> have been described .


It is evident that assembly of the surface domain of TF with VIIa to form the binary TF.VIIa complex induces a significant increase in the Kcat of the catalytic domain of VIIa for small peptidyl substrates and more profoundly for protein substrate .

It is evident that assembly of the <protein>surface domain</protein> of <protein>TF</protein> with <protein>VIIa</protein> to form the binary <protein>TF.VIIa complex</protein> induces a significant increase in the Kcat of the <protein>catalytic domain</protein> of <protein>VIIa</protein> for small peptidyl substrates and more profoundly for <protein>protein substrate</protein> .


This provides substantial evidence for an allosteric effect on the catalytic cleft of VIIa that is imparted by binding to TF , its cognate catalytic cofactor .

This provides substantial evidence for an allosteric effect on the catalytic cleft of <protein>VIIa</protein> that is imparted by binding to <protein>TF</protein> , its cognate catalytic <protein>cofactor</protein> .


It is also evident that the TF.VIIa complex is proteolytically active and can activate the zymogen plasma factor X to the serine protease Xa in free solution , inferring that extended substrate recognition by induced structural loci of the TF.VIIa complex are created from either or both proteins to constitute a new recognition structure .

It is also evident that the <protein>TF.VIIa complex</protein> is proteolytically active and can activate the <protein>zymogen plasma factor X</protein> to the <protein>serine protease Xa</protein> in free solution , inferring that extended substrate recognition by induced structural loci of the <protein>TF.VIIa complex</protein> are created from either or both proteins to constitute a <protein>new recognition structure</protein> .


It is also evident that association of X with charged phospholipid surfaces enhances the proteolytic activation of this zymogen by increasing recognition and susceptibility of the sessile peptide bond deduced from the markedly decreased Km and increased Kcat .

It is also evident that association of <protein>X</protein> with charged phospholipid surfaces enhances the proteolytic activation of this <protein>zymogen</protein> by increasing recognition and susceptibility of the sessile peptide bond deduced from the markedly decreased Km and increased Kcat .


Phorbol ester reduces constitutive nuclear NF kappa B and inhibits HIV-1 production in mature human monocytic cells .

Phorbol ester reduces <protein>constitutive nuclear NF kappa B</protein> and inhibits HIV-1 production in <cell_type>mature human monocytic cells</cell_type> .


NF kappa B is a potent mediator of specific gene expression in human monocytes and has been shown to play a role in transcription of the HIV-1 genome in promonocytic leukemias .

<protein>NF kappa B</protein> is a potent mediator of specific gene expression in human monocytes and has been shown to play a role in transcription of the HIV-1 genome in promonocytic leukemias .


There is little information available on the response of NF kappa B to cytokines in normal human monocytes .

There is little information available on the response of <protein>NF kappa B</protein> to <protein>cytokines</protein> in <cell_type>normal human monocytes</cell_type> .


We have used a 32P-labeled oligonucleotide derived from human immunodeficiency virus ( HIV-1 ) long terminal repeat , which contains a tandem repeat of the NF kappa B binding sequence , as a probe in a gel retardation assay to study this transcription factor .

We have used a 32P-labeled oligonucleotide derived from <dna>human immunodeficiency virus ( HIV-1 ) long terminal repeat</dna> , which contains a tandem repeat of the <dna>NF kappa B binding sequence</dna> , as a probe in a gel retardation assay to study this <protein>transcription factor</protein> .


Using this assay , we have detected NF kappa B in extracts of nuclei from normal human monocytes .

Using this assay , we have detected <protein>NF kappa B</protein> in extracts of nuclei from <cell_type>normal human monocytes</cell_type> .


Treatment of normal monocytes with 12-0-tetradecanoyl phorbol-13-acetate ( TPA ) for 4-24 h caused the complete disappearance of NF kappa B from nuclear extracts of monocytes .

Treatment of <cell_type>normal monocytes</cell_type> with 12-0-tetradecanoyl phorbol-13-acetate ( TPA ) for 4-24 h caused the complete disappearance of <protein>NF kappa B</protein> from nuclear extracts of monocytes .


A similar result was obtained with the mature monocytic leukemia cell line THP-1 .

A similar result was obtained with the <cell_line>mature monocytic leukemia cell line</cell_line> <cell_line>THP-1</cell_line> .


The constitutive transcription factor SP1 was unaffected by addition of TPA .

The <protein>constitutive transcription factor</protein> <protein>SP1</protein> was unaffected by addition of TPA .


The disappearance of NF kappa B from the nucleus was concentration dependent between 10 and 50 ng/ml of phorbol ester .

The disappearance of <protein>NF kappa B</protein> from the nucleus was concentration dependent between 10 and 50 ng/ml of phorbol ester .


In THP-1 cells , TPA also induced a new , faster-migrating NF kappa B species not induced in monocytes .

In THP-1 cells , TPA also induced a new , <protein>faster-migrating NF kappa B species</protein> not induced in <cell_type>monocytes</cell_type> .


Protein kinase C inhibitor staurosporine , but not cyclic nucleotide-dependent protein kinase inhibitor HA-1004 , also dramatically reduced constitutive levels of nuclear NF kappa B .

Protein kinase C inhibitor staurosporine , but not cyclic nucleotide-dependent protein kinase inhibitor HA-1004 , also dramatically reduced constitutive levels of <protein>nuclear NF kappa B</protein> .


Finally , TPA addition to monocytes infected with HIV-1 inhibited HIV-1 replication , as determined by reverse transcriptase assays , in a concentration-dependent manner .

Finally , TPA addition to <cell_type>monocytes</cell_type> infected with HIV-1 inhibited HIV-1 replication , as determined by <protein>reverse transcriptase</protein> assays , in a concentration-dependent manner .


These results are in striking contrast to the increase in nuclear NF kappa B and HIV-1 replication induced by phorbol esters in promonocytic leukemia cells U937 and HL-60 , and emphasize the importance of studying cytokine regulation of HIV-1 in normal monocytes .

These results are in striking contrast to the increase in nuclear <protein>NF kappa B</protein> and HIV-1 replication induced by phorbol esters in <cell_line>promonocytic leukemia cells U937</cell_line> and <cell_line>HL-60</cell_line> , and emphasize the importance of studying <protein>cytokine</protein> regulation of HIV-1 in <cell_type>normal monocytes</cell_type> .


A mechanism for the antiinflammatory effects of corticosteroids : the glucocorticoid receptor regulates leukocyte adhesion to endothelial cells and expression of endothelial-leukocyte adhesion molecule 1 and intercellular adhesion molecule 1 .

A mechanism for the antiinflammatory effects of corticosteroids : the <protein>glucocorticoid receptor</protein> regulates leukocyte adhesion to <cell_type>endothelial cells</cell_type> and expression of <protein>endothelial-leukocyte adhesion molecule 1</protein> and <protein>intercellular adhesion molecule 1</protein> .


Corticosteroids are the preeminent antiinflammatory agents although the molecular mechanisms that impart their efficacy have not been defined .


The endothelium plays a critical role in inflammation by directing circulating leukocytes into extravascular tissues by expressing adhesive molecules for leukocytes [ e.g. , endothelial-leukocyte adhesion molecule 1 ( ELAM-1 ) and intercellular adhesion molecule 1 ( ICAM-1 ) ] .

The endothelium plays a critical role in inflammation by directing <cell_type>circulating leukocytes</cell_type> into extravascular tissues by expressing <protein>adhesive molecules</protein> for <cell_type>leukocytes</cell_type> [ e.g. , <protein>endothelial-leukocyte adhesion molecule 1</protein> ( <protein>ELAM-1</protein> ) and <protein>intercellular adhesion molecule 1</protein> ( <protein>ICAM-1</protein> ) ] .


We therefore determined whether corticosteroids suppress inflammation by inhibiting endothelial expression of adhesion molecules for neutrophils ( polymorphonuclear leukocytes ) .

We therefore determined whether corticosteroids suppress inflammation by inhibiting endothelial expression of <protein>adhesion molecules</protein> for <cell_type>neutrophils</cell_type> ( <cell_type>polymorphonuclear leukocytes</cell_type> ) .


Preincubation of endothelial cells with endotoxin [ lipopolysaccharide ( LPS ) , 1 microgram/ml ] led to a 4-fold increase in subsequent adherence of polymorphonuclear leukocytes ( P < 0.0001 , n = 10 ) to endothelial cells , an increase that was markedly attenuated when endothelial cells were treated with dexamethasone ( IC50 < 1 nM , P < 0.0001 , n = 6 or 7 ) during preincubation with LPS .

Preincubation of <cell_type>endothelial cells</cell_type> with endotoxin [ lipopolysaccharide ( LPS ) , 1 microgram/ml ] led to a 4-fold increase in subsequent adherence of <cell_type>polymorphonuclear leukocytes</cell_type> ( P < 0.0001 , n = 10 ) to <cell_type>endothelial cells</cell_type> , an increase that was markedly attenuated when <cell_type>endothelial cells</cell_type> were treated with dexamethasone ( IC50 < 1 nM , P < 0.0001 , n = 6 or 7 ) during preincubation with LPS .


Moreover , the steroid receptor agonist cortisol ( 10 microM ) , but not its inactive metabolite tetrahydrocortisol ( 10 microM ) , diminished LPS-induced endothelial cell adhesiveness .

Moreover , the <protein>steroid receptor</protein> agonist cortisol ( 10 microM ) , but not its inactive metabolite tetrahydrocortisol ( 10 microM ) , diminished LPS-induced endothelial cell adhesiveness .


Further evidence that the action of dexamethasone was mediated through ligation of corticosteroid receptors [ human glucocorticoid receptors ( hGRs ) ] was provided by experiments utilizing the steroid antagonist RU-486 .

Further evidence that the action of dexamethasone was mediated through ligation of <protein>corticosteroid receptors</protein> [ <protein>human glucocorticoid receptors</protein> ( <protein>hGRs</protein> ) ] was provided by experiments utilizing the steroid antagonist RU-486 .


RU-486 ( 10 microM ) , which prevents translocation of ligated hGR to the nucleus by inhibiting dissociation of hGR from heat shock protein 90 , completely aborted the effect of dexamethasone on adhesiveness of endothelial cells ( P < 0.0005 , n = 3 ) .

RU-486 ( 10 microM ) , which prevents translocation of ligated <protein>hGR</protein> to the nucleus by inhibiting dissociation of <protein>hGR</protein> from heat shock protein 90 , completely aborted the effect of dexamethasone on adhesiveness of <cell_type>endothelial cells</cell_type> ( P < 0.0005 , n = 3 ) .


Treatment of endothelial cells with LPS ( 1 microgram/ml ) stimulated transcription of ELAM-1 , as shown by Northern blot analysis , and expression of membrane-associated ELAM-1 and ICAM-1 , as shown by quantitative immunofluorescence ( both P < 0.001 , n = 9 ) .

Treatment of <cell_type>endothelial cells</cell_type> with LPS ( 1 microgram/ml ) stimulated transcription of <protein>ELAM-1</protein> , as shown by Northern blot analysis , and expression of <protein>membrane-associated ELAM-1</protein> and <protein>ICAM-1</protein> , as shown by quantitative immunofluorescence ( both P < 0.001 , n = 9 ) .


Dexamethasone markedly inhibited LPS-stimulated accumulation of mRNA for ELAM-1 and expression of ELAM-1 and ICAM-1 ( IC50 < 10 nM , both P < 0.001 , n = 4-9 ) ; inhibition of expression by dexamethasone was reversed by RU-486 ( both P < 0.005 , n = 4-6 ) .

Dexamethasone markedly inhibited LPS-stimulated accumulation of mRNA for <protein>ELAM-1</protein> and expression of <protein>ELAM-1</protein> and <protein>ICAM-1</protein> ( IC50 < 10 nM , both P < 0.001 , n = 4-9 ) ; inhibition of expression by dexamethasone was reversed by RU-486 ( both P < 0.005 , n = 4-6 ) .


As in the adhesion studies , cortisol but not tetrahydrocortisol inhibited expression of ELAM-1 and ICAM-1 ( both P < 0.005 , n = 3 or 4 ) .

As in the adhesion studies , cortisol but not tetrahydrocortisol inhibited expression of <protein>ELAM-1</protein> and <protein>ICAM-1</protein> ( both P < 0.005 , n = 3 or 4 ) .


In contrast , sodium salicylate ( 1 mM ) inhibited neither adhesion nor expression of these adhesion molecules .

In contrast , sodium salicylate ( 1 mM ) inhibited neither adhesion nor expression of these <protein>adhesion molecules</protein> .


These studies suggest that antagonism by dexamethasone of endotoxin -induced inflammation is a specific instance of the general biological principle that the glucocorticoid receptor is a hormone-dependent regulator of transcription .

These studies suggest that antagonism by dexamethasone of <protein>endotoxin</protein> -induced inflammation is a specific instance of the general biological principle that the <protein>glucocorticoid receptor</protein> is a hormone-dependent regulator of transcription .


Pax-5 encodes the transcription factor BSAP and is expressed in B lymphocytes , the developing CNS , and adult testis .

<dna>Pax-5</dna> encodes the <protein>transcription factor</protein> <protein>BSAP</protein> and is expressed in <cell_type>B lymphocytes</cell_type> , the developing CNS , and adult testis .


BSAP has been identified previously as a transcription factor that is expressed at early , but not late , stages of B-cell differentiation .

<protein>BSAP</protein> has been identified previously as a <protein>transcription factor</protein> that is expressed at early , but not late , stages of B-cell differentiation .


Biochemical purification and cDNA cloning has now revealed that BSAP belongs to the family of paired domain proteins .

Biochemical purification and cDNA cloning has now revealed that <protein>BSAP</protein> belongs to the family of <protein>paired domain proteins</protein> .


BSAP is encoded by the Pax-5 gene and has been highly conserved between human and mouse .

<protein>BSAP</protein> is encoded by the <dna>Pax-5 gene</dna> and has been highly conserved between human and mouse .


An intact paired domain was shown to be both necessary and sufficient for DNA binding of BSAP .

An intact paired domain was shown to be both necessary and sufficient for DNA binding of <protein>BSAP</protein> .


Binding studies with several BSAP recognition sequences demonstrated that the sequence specificity of BSAP differs from that of the distantly related paired domain protein Pax-1 .

Binding studies with several <dna>BSAP recognition sequences</dna> demonstrated that the sequence specificity of <protein>BSAP</protein> differs from that of the distantly related <protein>paired domain protein Pax-1</protein> .


During embryogenesis , the BSAP gene is transiently expressed in the mesencephalon and spinal cord with a spatial and temporal expression pattern that is distinct from that of other Pax genes in the developing central nervous system ( CNS ) .

During embryogenesis , the <dna>BSAP gene</dna> is transiently expressed in the mesencephalon and spinal cord with a spatial and temporal expression pattern that is distinct from that of other <dna>Pax genes</dna> in the developing central nervous system ( CNS ) .


Later , the expression of the BSAP gene shifts to the fetal liver where it correlates with the onset of B lymphopoiesis .

Later , the expression of the <dna>BSAP gene</dna> shifts to the fetal liver where it correlates with the onset of B lymphopoiesis .


BSAP expression persists in B lymphocytes and is also seen in the testis of the adult mouse .

<protein>BSAP</protein> expression persists in <cell_type>B lymphocytes</cell_type> and is also seen in the testis of the adult mouse .


All of this evidence indicates that the transcription factor BSAP may not only play an important role in B-cell differentiation but also in neural development and spermatogenesis .

All of this evidence indicates that the <protein>transcription factor</protein> <protein>BSAP</protein> may not only play an important role in B-cell differentiation but also in neural development and spermatogenesis .


Cell cycle-dependent initiation and lineage-dependent abrogation of GATA-1 expression in pure differentiating hematopoietic progenitors .

Cell cycle-dependent initiation and lineage-dependent abrogation of <protein>GATA-1</protein> expression in <cell_type>pure differentiating hematopoietic progenitors</cell_type> .


The programmed activation/repression of transcription factors in early hematopoietic differentiation has not yet been explored .

The programmed activation/repression of <protein>transcription factors</protein> in early hematopoietic differentiation has not yet been explored .


The DNA-binding protein GATA-1 is required for normal erythroid development and regulates erythroid-expressed genes in maturing erythroblasts .

The <protein>DNA-binding protein</protein> <protein>GATA-1</protein> is required for normal erythroid development and regulates <dna>erythroid-expressed genes</dna> in <cell_type>maturing erythroblasts</cell_type> .


We analyzed GATA-1 expression in early human adult hematopoiesis by using an in vitro system in which `` pure '' early hematopoietic progenitors are induced to gradual and synchronized differentiation selectively along the erythroid or granulocyte-macrophage pathway by differential treatment with hematopoietic growth factors .

We analyzed <protein>GATA-1</protein> expression in early human adult hematopoiesis by using an in vitro system in which `` pure '' <cell_type>early hematopoietic progenitors</cell_type> are induced to gradual and synchronized differentiation selectively along the erythroid or granulocyte-macrophage pathway by differential treatment with hematopoietic growth factors .


The GATA-1 gene , though virtually silent in quiescent progenitors , is activated after entrance into the cell cycle upon stimulation with hematopoietic growth factors .

The <dna>GATA-1 gene</dna> , though virtually silent in <cell_type>quiescent progenitors</cell_type> , is activated after entrance into the cell cycle upon stimulation with hematopoietic growth factors .


Subsequently , increasing expression along the erythroid pathway contrasts with an abrupt downregulation in the granulocyte-macrophage lineage .

Subsequently , increasing expression along the erythroid pathway contrasts with an abrupt downregulation in the <cell_type>granulocyte-macrophage lineage</cell_type> .


These results suggest a microenvironment-directed , two-step model for GATA-1 expression in differentiating hematopoietic progenitors that involves ( i ) cycle-dependent initiation and ( ii ) lineage-dependent maintenance or suppression .

These results suggest a microenvironment-directed , two-step model for <protein>GATA-1</protein> expression in <cell_type>differentiating hematopoietic progenitors</cell_type> that involves ( i ) cycle-dependent initiation and ( ii ) lineage-dependent maintenance or suppression .


Hypothetically , on/off switches of lineage-restricted transactivators may underlie the binary fate decisions of hematopoietic progenitors .

Hypothetically , on/off switches of <protein>lineage-restricted transactivators</protein> may underlie the binary fate decisions of <cell_type>hematopoietic progenitors</cell_type> .


Specific NF-kappa B subunits act in concert with Tat to stimulate human immunodeficiency virus type 1 transcription .

Specific <protein>NF-kappa B subunits</protein> act in concert with <protein>Tat</protein> to stimulate human immunodeficiency virus type 1 transcription .


NF-kappa B is a protein complex which functions in concert with the tat-I gene product to stimulate human immunodeficiency virus ( HIV ) transcription .

<protein>NF-kappa B</protein> is a <protein>protein complex</protein> which functions in concert with the <protein>tat-I gene product</protein> to stimulate human immunodeficiency virus ( HIV ) transcription .


To determine whether specific members of the NF-kappa B family contribute to this effect , we have examined the abilities of different NF-kappa B subunits to act with Tat-I to stimulate transcription of HIV in Jurkat T-leukemia cells .

To determine whether specific members of the <protein>NF-kappa B family</protein> contribute to this effect , we have examined the abilities of different <protein>NF-kappa B subunits</protein> to act with <protein>Tat-I</protein> to stimulate transcription of HIV in <cell_type>Jurkat T-leukemia cells</cell_type> .


We have found that the p49 ( 100 ) DNA binding subunit , together with p65 , can act in concert with Tat-I to stimulate the expression of HIV-CAT plasmid .

We have found that the <protein>p49 ( 100 ) DNA binding subunit</protein> , together with <protein>p65</protein> , can act in concert with <protein>Tat-I</protein> to stimulate the expression of <dna>HIV-CAT plasmid</dna> .


Little effect was observed with 50-kDa forms of p105 NF-kappa B or rel , in combination with p65 or full-length c-rel , which do not stimulate the HIV enhancer in these cells .

Little effect was observed with <protein>50-kDa forms</protein> of <protein>p105</protein> <protein>NF-kappa B</protein> or <protein>rel</protein> , in combination with <protein>p65</protein> or <protein>full-length c-rel</protein> , which do not stimulate the <dna>HIV enhancer</dna> in these cells .


These findings suggest that the combination of p49 ( 100 ) and p65 NF-kappa B can act in concert with the tat-I gene product to stimulate the synthesis of HIV RNA .

These findings suggest that the combination of <protein>p49 ( 100 )</protein> and <protein>p65</protein> <protein>NF-kappa B</protein> can act in concert with the <protein>tat-I gene product</protein> to stimulate the synthesis of <rna>HIV RNA</rna> .


Stable expression of HB24 , a diverged human homeobox gene , in T lymphocytes induces genes involved in T cell activation and growth .

Stable expression of <dna>HB24</dna> , a <dna>diverged human homeobox gene</dna> , in T lymphocytes induces genes involved in T cell activation and growth .


A diverged homeobox gene , HB24 , which is known to be induced following lymphocyte activation , was introduced into Jurkat T cells under the control of a constitutive promoter .

A <dna>diverged homeobox gene</dna> , <dna>HB24</dna> , which is known to be induced following lymphocyte activation , was introduced into <cell_line>Jurkat T cells</cell_line> under the control of a <dna>constitutive promoter</dna> .


Stable transfectants of HB24 were established that expressed high levels of HB24 mRNA and possessed an altered phenotype suggestive of activated T cells .

Stable transfectants of <dna>HB24</dna> were established that expressed high levels of <rna>HB24 mRNA</rna> and possessed an altered phenotype suggestive of <cell_type>activated T cells</cell_type> .


A number of genes known to be induced following T cell activation and associated with cell growth were increased in the transfectants , including c-fos , c-myc , c-myb , HLA-DR , lck , NF-kappa B , interleukin-2 and interleukin-2 receptor alpha ( IL-2R alpha ) .

A number of genes known to be induced following T cell activation and associated with cell growth were increased in the <cell_line>transfectants</cell_line> , including <dna>c-fos</dna> , <dna>c-myc</dna> , <dna>c-myb</dna> , <dna>HLA-DR</dna> , <dna>lck</dna> , <protein>NF-kappa B</protein> , <dna>interleukin-2</dna> and <dna>interleukin-2 receptor alpha</dna> ( <dna>IL-2R alpha</dna> ) .


Analysis of IL-2R alpha expression by transient transfection of IL-2R alpha promoter constructs into the HB24 transfectants revealed constitutive expression ( about 60 % of phytohemagglutinin- and phorbol ester-activated Jurkat cells ) that was dependent on the kappa B site in the IL-2R alpha promoter .

Analysis of <protein>IL-2R alpha</protein> expression by transient transfection of <dna>IL-2R alpha promoter constructs</dna> into the <cell_line>HB24 transfectants</cell_line> revealed constitutive expression ( about 60 % of <cell_line>phytohemagglutinin- and phorbol ester-activated Jurkat cells</cell_line> ) that was dependent on the <dna>kappa B site</dna> in the <dna>IL-2R alpha promoter</dna> .


Furthermore , as a consequence of the increased HB24 mRNA levels , the Jurkat HB24 transfectants proliferated more rapidly than control cell lines .

Furthermore , as a consequence of the increased <rna>HB24 mRNA</rna> levels , the <cell_line>Jurkat HB24 transfectants</cell_line> proliferated more rapidly than <cell_line>control cell lines</cell_line> .


Thus , stable expression of HB24 confers an activation phenotype on a human T cell line , implicating this gene as an important transcriptional factor during T cell activation and growth .

Thus , stable expression of <dna>HB24</dna> confers an activation phenotype on a <cell_line>human T cell line</cell_line> , implicating this gene as an important <protein>transcriptional factor</protein> during T cell activation and growth .


Functional interaction between the two zinc finger domains of the v-erb A oncoprotein .

Functional interaction between the two <protein>zinc finger domains</protein> of the <protein>v-erb A oncoprotein</protein> .


The v-erb A oncogene of avian erythroblastosis virus is a mutated and virally transduced copy of a host cell gene encoding a thyroid hormone receptor .

The <dna>v-erb A oncogene</dna> of avian erythroblastosis virus is a mutated and virally transduced copy of a <dna>host cell gene</dna> encoding a <protein>thyroid hormone receptor</protein> .


The protein expressed by the v-erb A oncogene binds to DNA and acts as a dominant negative inhibitor of both the thyroid hormone receptor and the closely related retinoic acid receptor .

The protein expressed by the <dna>v-erb A oncogene</dna> binds to DNA and acts as a dominant negative inhibitor of both the <protein>thyroid hormone receptor</protein> and the closely related <protein>retinoic acid receptor</protein> .


The v-erb A protein has sustained two amino acid alterations within its DNA-binding domain relative to that of c-erb A , one of which , at serine 61 , is known to be important for v-erb A function in the neoplastic cell .

The <protein>v-erb A protein</protein> has sustained two amino acid alterations within its <protein>DNA-binding domain</protein> relative to that of <protein>c-erb A</protein> , one of which , at serine 61 , is known to be important for <protein>v-erb A</protein> function in the <cell_type>neoplastic cell</cell_type> .


We report here that the second alteration , at threonine 78 , also plays an important , although more indirect , role : alteration of the sequence at threonine 78 such that it resembles that of c-erb A can act as an intragenic suppressor and can partially restore function to a v-erb A protein rendered defective due to a mutation at position 61 .

We report here that the second alteration , at threonine 78 , also plays an important , although more indirect , role : alteration of the sequence at threonine 78 such that it resembles that of <protein>c-erb A</protein> can act as an intragenic suppressor and can partially restore function to a <protein>v-erb A protein</protein> rendered defective due to a mutation at <protein>position 61</protein> .


Threonine 78 lies within the D-box of the v-erb A protein , a region thought to mediate receptor-receptor dimerizations , and is not in physical proximity to the serine at position 61 .

Threonine 78 lies within the <protein>D-box</protein> of the <protein>v-erb A protein</protein> , a region thought to mediate receptor-receptor dimerizations , and is not in physical proximity to the serine at <protein>position 61</protein> .


It therefore appears that an indirect interaction occurs between these two sites and that this interaction is crucial for v-erb A function .

It therefore appears that an indirect interaction occurs between these two sites and that this interaction is crucial for <protein>v-erb A</protein> function .


cis-acting sequences required for inducible interleukin-2 enhancer function bind a novel Ets-related protein , Elf-1 .

<dna>cis-acting sequences</dna> required for inducible <dna>interleukin-2 enhancer</dna> function bind a novel <protein>Ets-related protein</protein> , <protein>Elf-1</protein> .


The recent definition of a consensus DNA binding sequence for the Ets family of transcription factors has allowed the identification of potential Ets binding sites in the promoters and enhancers of many inducible T-cell genes .

The recent definition of a consensus <dna>DNA binding sequence</dna> for the <protein>Ets family</protein> of <protein>transcription factors</protein> has allowed the identification of potential <dna>Ets binding sites</dna> in the <dna>promoters</dna> and <dna>enhancers</dna> of many <dna>inducible T-cell genes</dna> .


In the studies described in this report , we have identified two potential Ets binding sites , EBS1 and EBS2 , which are conserved in both the human and murine interleukin-2 enhancers .

In the studies described in this report , we have identified two potential <dna>Ets binding sites</dna> , EBS1 and <dna>EBS2</dna> , which are conserved in both the <dna>human and murine interleukin-2 enhancers</dna> .


Within the human enhancer , these two sites are located within the previously defined DNase I footprints , NFAT-1 and NFIL-2B , respectively .

Within the <dna>human enhancer</dna> , these two sites are located within the previously defined <protein>DNase I</protein> footprints , <dna>NFAT-1</dna> and <dna>NFIL-2B</dna> , respectively .


Electrophoretic mobility shift and methylation interference analyses demonstrated that EBS1 and EBS2 are essential for the formation of the NFAT-1 and NFIL-2B nuclear protein complexes .

Electrophoretic mobility shift and methylation interference analyses demonstrated that EBS1 and <dna>EBS2</dna> are essential for the formation of the <protein>NFAT-1 and NFIL-2B nuclear protein complexes</protein> .


Furthermore , in vitro mutagenesis experiments demonstrated that inducible interleukin-2 enhancer function requires the presence of either EBS1 or EBS2 .

Furthermore , in vitro mutagenesis experiments demonstrated that inducible <dna>interleukin-2 enhancer</dna> function requires the presence of either EBS1 or <dna>EBS2</dna> .


Two well-characterized Ets family members , Ets-1 and Ets-2 , are reciprocally expressed during T-cell activation .

Two well-characterized <protein>Ets family members</protein> , <protein>Ets-1</protein> and <protein>Ets-2</protein> , are reciprocally expressed during T-cell activation .


Surprisingly , however , neither of these proteins bound in vitro to EBS1 or EBS2 .

Surprisingly , however , neither of these proteins bound in vitro to EBS1 or <dna>EBS2</dna> .


We therefore screened a T-cell cDNA library under low-stringency conditions with a probe from the DNA binding domain of Ets-1 and isolated a novel Ets family member , Elf-1 .

We therefore screened a <dna>T-cell cDNA</dna> library under low-stringency conditions with a probe from the <protein>DNA binding domain</protein> of <protein>Ets-1</protein> and isolated a <protein>novel Ets family member</protein> , <protein>Elf-1</protein> .


Elf-1 contains a DNA binding domain that is nearly identical to that of E74 , the ecdysone-inducible Drosophila transcription factor required for metamorphosis ( hence the name Elf-1 , for E74-like factor 1 ) .

<protein>Elf-1</protein> contains a <protein>DNA binding domain</protein> that is nearly identical to that of <protein>E74</protein> , the <protein>ecdysone-inducible Drosophila transcription factor</protein> required for metamorphosis ( hence the name <protein>Elf-1</protein> , for <protein>E74-like factor 1</protein> ) .


Elf-1 bound specifically to both EBS1 and EBS2 in electrophoretic mobility shift assays .

<protein>Elf-1</protein> bound specifically to both EBS1 and <dna>EBS2</dna> in electrophoretic mobility shift assays .


It also bound to the purine-rich CD3R element from the human immunodeficiency virus type 2 long terminal repeat , which is required for inducible virus expression in response to signalling through the T-cell receptor .

It also bound to the <dna>purine-rich CD3R element</dna> from the <dna>human immunodeficiency virus type 2 long terminal repeat</dna> , which is required for inducible virus expression in response to signalling through the <protein>T-cell receptor</protein> .


Taken together , these results demonstrate that multiple Ets family members with apparently distinct DNA binding specificities regulate differential gene expression in resting and activated T cells .

Taken together , these results demonstrate that <protein>multiple Ets family members</protein> with apparently distinct DNA binding specificities regulate differential gene expression in <cell_type>resting and activated T cells</cell_type> .


Binding of erythroid and non-erythroid nuclear proteins to the silencer of the human epsilon-globin-encoding gene .

Binding of <protein>erythroid and non-erythroid nuclear proteins</protein> to the <dna>silencer</dna> of the <dna>human epsilon-globin-encoding gene</dna> .


To clarify the molecular mechanisms involved in the developmental control of hemoglobin-encoding genes we have been studying the expression of these genes in human cells in continuous culture .

To clarify the molecular mechanisms involved in the developmental control of <dna>hemoglobin-encoding genes</dna> we have been studying the expression of these <dna>genes</dna> in <cell_type>human cells</cell_type> in continuous culture .


We have previously reported the presence of a transcriptional control element with the properties of a silencer extending from -392 to -177 bp relative to the cap site of the human epsilon-globin-encoding gene [ Cao et al. , Proc.Natl.Acad.Sci.USA 86 ( 1989 ) 5306-5309 ] .

We have previously reported the presence of a <dna>transcriptional control element</dna> with the properties of a <dna>silencer</dna> extending from <dna>-392 to -177 bp</dna> relative to the <dna>cap site</dna> of the <dna>human epsilon-globin-encoding gene</dna> [ Cao et al. , Proc.Natl.Acad.Sci.USA 86 ( 1989 ) 5306-5309 ] .


We also showed that this silencer has stronger inhibitory activity in HeLa cells , as compared to K562 human erythroleukemia cells .

We also showed that this <dna>silencer</dna> has stronger inhibitory activity in <cell_line>HeLa cells</cell_line> , as compared to <cell_line>K562 human erythroleukemia cells</cell_line> .


Using deletion mutants and cis-cloned synthetic oligodeoxyribonucleotides in transient expression assays , nucleotide sequences responsible for this effect have now been further delimited to 44 bp located from -294 to -251 bp .

Using <dna>deletion mutants</dna> and cis-cloned synthetic oligodeoxyribonucleotides in transient expression assays , nucleotide sequences responsible for this effect have now been further delimited to 44 bp located from <dna>-294 to -251 bp</dna> .


Gel electrophoresis mobility shift assays and DNaseI footprinting assays demonstrate that these negative regulatory sequences are recognized differently by proteins present in nuclear extracts obtained from HeLa and K562 cells .

Gel electrophoresis mobility shift assays and <protein>DNaseI</protein> footprinting assays demonstrate that these negative regulatory sequences are recognized differently by proteins present in nuclear extracts obtained from <cell_line>HeLa</cell_line> and <cell_line>K562 cells</cell_line> .


Two binding proteins are detected in K562 nuclear extracts , while only one is found in extracts from HeLa cells .

Two binding proteins are detected in K562 nuclear extracts , while only one is found in extracts from <cell_line>HeLa cells</cell_line> .


Possible mechanisms by which these proteins may regulate transcription of the epsilon-globin-encoding gene in erythroid and non-erythroid cells are discussed .

Possible mechanisms by which these proteins may regulate transcription of the <dna>epsilon-globin-encoding gene</dna> in <cell_type>erythroid and non-erythroid cells</cell_type> are discussed .


Calcitriol : a hematolymphopoietrope ? [ editorial ]


A MEDLINE search of the English-language literature was conducted using the indexing terms 'immunology , calcitriol and vitamin D ' to identify studies indicating a role for calcitriol as a primary immunomodulator .


Sixty-six papers published between January 1956 and June 1991 were identified .


Forty-five of these reports are cited in this review .


The data strongly suggest an endocrine , autocrine and/or paracrine role for calcitriol in immune regulation .


No unifying hypothesis has yet emerged explaining this collection of data .


This paper provides a brief review of immune properties currently attributed to calcitriol .


Transcription of the hypersensitive site HS2 enhancer in erythroid cells .

Transcription of the <dna>hypersensitive site HS2 enhancer</dna> in <cell_type>erythroid cells</cell_type> .


In the human genome , the erythroid-specific hypersensitive site HS2 enhancer regulates the transcription of the downstream beta-like globin genes 10-50 kilobases away .

In the <dna>human genome</dna> , the <dna>erythroid-specific hypersensitive site HS2 enhancer</dna> regulates the transcription of the <dna>downstream beta-like globin genes</dna> <dna>10-50 kilobases</dna> away .


The mechanism of HS2 enhancer function is not known .

The mechanism of <dna>HS2 enhancer</dna> function is not known .


The present study employs RNA protection assays to analyze the transcriptional status of the HS2 enhancer in transfected recombinant chloramphenicol acetyltransferase ( CAT ) plasmids .

The present study employs RNA protection assays to analyze the transcriptional status of the <dna>HS2 enhancer</dna> in <dna>transfected recombinant chloramphenicol acetyltransferase ( CAT ) plasmids</dna> .


In erythroid K562 cells in which the HS2 enhancer is active , the HS2 sequence directs the synthesis of long enhancer transcripts that are initiated apparently from within the enhancer and elongated through the intervening DNA into the cis-linked CAT gene .

In <cell_line>erythroid K562 cells</cell_line> in which the <dna>HS2 enhancer</dna> is active , the <dna>HS2 sequence</dna> directs the synthesis of long enhancer transcripts that are initiated apparently from within the <dna>enhancer</dna> and elongated through the intervening DNA into the <dna>cis-linked CAT gene</dna> .


In nonerythroid HL-60 cells in which the HS2 enhancer is inactive , long enhancer transcripts are not detectable .

In <cell_line>nonerythroid HL-60 cells</cell_line> in which the <dna>HS2 enhancer</dna> is inactive , <rna>long enhancer transcripts</rna> are not detectable .


Splitting the HS2 enhancer between two tandem Ap1 sites abolishes the synthesis of a group of long enhancer transcripts and results in loss of enhancer function and transcriptional silencing of the cis-linked CAT gene .

Splitting the <dna>HS2 enhancer</dna> between two <dna>tandem Ap1 sites</dna> abolishes the synthesis of a group of <rna>long enhancer transcripts</rna> and results in loss of enhancer function and transcriptional silencing of the cis-linked CAT gene .


In directing the synthesis of RNA through the intervening DNA and the gene by a tracking and transcription mechanism , the HS2 enhancer may ( i ) open up the chromatin structure of a gene domain and ( ii ) deliver enhancer binding proteins to the promoter sequence where they may stimulate the transcription of the gene at the cap site .

In directing the synthesis of <rna>RNA</rna> through the intervening <dna>DNA</dna> and the gene by a tracking and transcription mechanism , the <dna>HS2 enhancer</dna> may ( i ) open up the <dna>chromatin structure</dna> of a <dna>gene domain</dna> and ( ii ) deliver <protein>enhancer binding proteins</protein> to the promoter sequence where they may stimulate the transcription of the gene at the <dna>cap site</dna> .


Characterization of a novel T lymphocyte protein which binds to a site related to steroid/thyroid hormone receptor response elements in the negative regulatory sequence of the human immunodeficiency virus long terminal repeat .

Characterization of a <protein>novel T lymphocyte protein</protein> which binds to a site related to <dna>steroid/thyroid hormone receptor response elements</dna> in the <dna>negative regulatory sequence</dna> of the <dna>human immunodeficiency virus long terminal repeat</dna> .


We have previously identified a T lymphocyte protein which binds to a site within the LTR of the human immunodeficiency virus type 1 ( HIV-1 ) and exerts an inhibitory effect on virus gene expression .

We have previously identified a <protein>T lymphocyte protein</protein> which binds to a site within the <dna>LTR</dna> of the human immunodeficiency virus type 1 ( HIV-1 ) and exerts an inhibitory effect on virus gene expression .


The palindromic site ( site B ) recognized by this protein is related to the palindromic binding sites of members of the steroid/thyroid hormone receptor family .

The <dna>palindromic site</dna> ( <dna>site B</dna> ) recognized by this protein is related to the <dna>palindromic binding sites</dna> of members of the <protein>steroid/thyroid hormone receptor family</protein> .


Here we characterize the T cell protein binding to this site as a 100 kD protein which is most abundant in T cells and which binds to site B as a 200 kD complex .

Here we characterize the <protein>T cell protein</protein> binding to this site as a 100 kD protein which is most abundant in <cell_type>T cells</cell_type> and which binds to <dna>site B</dna> as a <protein>200 kD complex</protein> .


This protein is distinct from other members of the steroid/thyroid hormone receptor family including the COUP protein which has a closely related DNA binding specificity .

This protein is distinct from other members of the <protein>steroid/thyroid hormone receptor family</protein> including the <protein>COUP protein</protein> which has a closely related DNA binding specificity .


TAR -independent transactivation by Tat in cells derived from the CNS : a novel mechanism of HIV-1 gene regulation .

<dna>TAR</dna> -independent transactivation by <protein>Tat</protein> in cells derived from the CNS : a novel mechanism of HIV-1 gene regulation .


The Tat protein of human immunodeficiency virus type 1 ( HIV-1 ) is essential for productive infection and is a potential target for antiviral therapy .

The <protein>Tat protein</protein> of human immunodeficiency virus type 1 ( HIV-1 ) is essential for productive infection and is a potential target for antiviral therapy .


Tat , a potent activator of HIV-1 gene expression , serves to greatly increase the rate of transcription directed by the viral promoter .

<protein>Tat</protein> , a potent activator of HIV-1 gene expression , serves to greatly increase the rate of transcription directed by the <dna>viral promoter</dna> .


This induction , which seems to be an important component in the progression of acquired immune deficiency syndrome ( AIDS ) , may be due to increased transcriptional initiation , increased transcriptional elongation , or a combination of these processes .


Much attention has been focused on the interaction of Tat with a specific RNA target termed TAR ( transactivation responsive ) which is present in the leader sequence of all HIV-1 mRNAs .

Much attention has been focused on the interaction of <protein>Tat</protein> with a specific <rna>RNA target</rna> termed <rna>TAR</rna> ( <rna>transactivation responsive</rna> ) which is present in the leader sequence of all <rna>HIV-1 mRNAs</rna> .


This interaction is believed to be an important component of the mechanism of transactivation .


In this report we demonstrate that in certain CNS-derived cells Tat is capable of activating HIV-1 through a TAR -independent pathway .

In this report we demonstrate that in certain <cell_line>CNS-derived cells</cell_line> <protein>Tat</protein> is capable of activating HIV-1 through a <dna>TAR</dna> -independent pathway .


A Tat-responsive element is found upstream within the viral promoter that in glial-derived cell lines allows transactivation in the absence of TAR .

A <dna>Tat-responsive element</dna> is found upstream within the <dna>viral promoter</dna> that in <cell_line>glial-derived cell lines</cell_line> allows transactivation in the absence of <rna>TAR</rna> .


Deletion mapping and hybrid promoter constructs demonstrate that the newly identified Tat-responsive element corresponds to a sequence within the viral long terminal repeat ( LTR ) previously identified as the HIV-1 enhancer , or NF-kappa B domain .

Deletion mapping and <dna>hybrid promoter constructs</dna> demonstrate that the newly identified <dna>Tat-responsive element</dna> corresponds to a sequence within the <dna>viral long terminal repeat</dna> ( <dna>LTR</dna> ) previously identified as the <dna>HIV-1 enhancer</dna> , or <dna>NF-kappa B domain</dna> .


DNA band-shift analysis reveals NF-kappa B binding activity in glial cells that differs from that present in T lymphoid cells .

DNA band-shift analysis reveals <protein>NF-kappa B</protein> binding activity in glial cells that differs from that present in <cell_type>T lymphoid cells</cell_type> .


Further , we observe that TAR-deleted mutants of HIV-1 demonstrate normal late gene expression in glial cells as evidenced by syncytia formation and production of viral p24 antigen .

Further , we observe that <dna>TAR-deleted mutants</dna> of HIV-1 demonstrate normal late gene expression in <cell_type>glial cells</cell_type> as evidenced by syncytia formation and production of <protein>viral p24 antigen</protein> .


( ABSTRACT TRUNCATED AT 250 WORDS )


Transcription factor AP-2 activates gene expression of HTLV-I .

<protein>Transcription factor AP-2</protein> activates gene expression of HTLV-I .


The HTLV-I LTR contains three conserved regulatory elements known as 21 base pair repeats which are required for stimulation of gene expression by the transactivator protein tax .

The <dna>HTLV-I LTR</dna> contains three conserved <dna>regulatory elements</dna> known as <dna>21 base pair repeats</dna> which are required for stimulation of gene expression by the <protein>transactivator protein</protein> <protein>tax</protein> .


Mutagenesis indicates that the 21 bp repeats can be subdivided into three motifs , A , B and C , each of which influences the level of tax activation .

Mutagenesis indicates that the <dna>21 bp repeats</dna> can be subdivided into three motifs , A , B and C , each of which influences the level of <protein>tax</protein> activation .


The A site in the 21 bp repeat has strong homology with previously described binding sites for the transcription factor AP-2 .

The <dna>A site</dna> in the <dna>21 bp repeat</dna> has strong homology with previously described <dna>binding sites</dna> for the <protein>transcription factor AP-2</protein> .


We demonstrated that AP-2 mRNA was present in T-lymphocytes and that cellular factors from both non-transformed and transformed T-lymphocytes specifically bound to the consensus motif for AP-2 in each 21 bp .

We demonstrated that <rna>AP-2 mRNA</rna> was present in <cell_type>T-lymphocytes</cell_type> and that cellular factors from both non-transformed and transformed <cell_type>T-lymphocytes</cell_type> specifically bound to the <dna>consensus motif</dna> for <protein>AP-2</protein> in each 21 bp .


To determine the role of AP-2 in the regulation of the HTLV-I LTR gene expression , we used an AP-2 cDNA in DNA binding and transient expression assays .

To determine the role of <protein>AP-2</protein> in the regulation of the <dna>HTLV-I LTR</dna> gene expression , we used an <dna>AP-2 cDNA</dna> in DNA binding and transient expression assays .


Gel retardation and methylation interference studies revealed that bacterially produced AP-2 bound specifically and with high affinity to all three 21 bp repeats , and that it required the core sequence AGGC for specific binding .

Gel retardation and methylation interference studies revealed that <protein>bacterially produced AP-2</protein> bound specifically and with high affinity to all three <dna>21 bp repeats</dna> , and that it required the <dna>core sequence</dna> AGGC for specific binding .


Binding of AP-2 prevented the subsequent binding of members of the CREB/ATF family to an adjacent regulatory motif in the 21 bp repeat .

Binding of <protein>AP-2</protein> prevented the subsequent binding of members of the <protein>CREB/ATF family</protein> to an <dna>adjacent regulatory motif</dna> in the <dna>21 bp repeat</dna> .


Transfection of an AP-2 expression construct into T-lymphocytes activated gene expression from the HTLV-I LTR .

Transfection of an <dna>AP-2 expression construct</dna> into <cell_type>T-lymphocytes</cell_type> activated gene expression from the <dna>HTLV-I LTR</dna> .


At least two 21 bp repeats were required for high levels of AP-2 activation and mutagenesis of the AP-2 consensus binding sequences in the 21 bp repeats eliminate this activation .

At least two <dna>21 bp repeats</dna> were required for high levels of <protein>AP-2</protein> activation and mutagenesis of the <dna>AP-2 consensus binding sequences</dna> in the <dna>21 bp repeats</dna> eliminate this activation .


Activation of the human immunodeficiency virus type 1 enhancer is not dependent on NFAT-1 .

Activation of the <dna>human immunodeficiency virus type 1 enhancer</dna> is not dependent on <protein>NFAT-1</protein> .


The function of a putative NFAT-1 site in the human immunodeficiency virus type 1 enhancer has been analyzed .

The function of a <dna>putative NFAT-1 site</dna> in the <dna>human immunodeficiency virus type 1 enhancer</dna> has been analyzed .


Activation by the T-cell antigen receptor is minimal in Jurkat cells and is mediated by the kappa B sites .

Activation by the <protein>T-cell antigen receptor</protein> is minimal in <cell_line>Jurkat cells</cell_line> and is mediated by the <dna>kappa B sites</dna> .


The putative NFAT-1 region is not required for the response to anti-CD3 or to mitogens in T-cell , B-cell , or monocyte/macrophage leukemia lines , nor is it a cis-acting negative regulatory element .

The <dna>putative NFAT-1 region</dna> is not required for the response to <protein>anti-CD3</protein> or to <protein>mitogens</protein> in <cell_line>T-cell</cell_line> , <cell_line>B-cell</cell_line> , or <cell_line>monocyte/macrophage leukemia lines</cell_line> , nor is it a <dna>cis-acting negative regulatory element</dna> .


The B cell-specific nuclear factor OTF-2 positively regulates transcription of the human class II transplantation gene , DRA .

The <protein>B cell-specific nuclear factor</protein> <protein>OTF-2</protein> positively regulates transcription of the <dna>human class II transplantation gene</dna> , <dna>DRA</dna> .


The promoter of the major histocompatibility class II gene DRA contains an octamer element ( ATTTGCAT ) that is required for efficient DRA expression in B cells .

The promoter of the <dna>major histocompatibility class II gene</dna> <dna>DRA</dna> contains an <dna>octamer element</dna> ( ATTTGCAT ) that is required for efficient <dna>DRA</dna> expression in <cell_type>B cells</cell_type> .


Several DNA-binding proteins are known to bind this sequence .

Several <protein>DNA-binding proteins</protein> are known to bind this sequence .


The best characterized are the B cell-specific OTF-2 and the ubiquitous OTF-1 .

The best characterized are the <protein>B cell-specific OTF-2</protein> and the ubiquitous <protein>OTF-1</protein> .


This report directly demonstrates that OTF-2 but not OTF-1 regulates the DRA gene .

This report directly demonstrates that <protein>OTF-2</protein> but not <protein>OTF-1</protein> regulates the <dna>DRA gene</dna> .


In vitro transcription analysis using protein fractions enriched for the octamer-binding protein OTF-2 demonstrate a positive functional role for OTF-2 in DRA gene transcription .

In vitro transcription analysis using protein fractions enriched for the octamer-binding protein <protein>OTF-2</protein> demonstrate a positive functional role for <protein>OTF-2</protein> in <dna>DRA gene</dna> transcription .


In contrast , OTF-1 -enriched protein fractions did not affect DRA gene transcription although it functionally enhanced the transcription of another gene .

In contrast , <protein>OTF-1</protein> -enriched protein fractions did not affect <dna>DRA gene</dna> transcription although it functionally enhanced the transcription of another gene .


Recombinant OTF-2 protein produced by in vitro transcription/translation could also enhance DRA gene transcription in vitro .

<protein>Recombinant OTF-2 protein</protein> produced by in vitro transcription/translation could also enhance <dna>DRA gene</dna> transcription in vitro .


In vivo transient transfection studies utilizing an OTF-2 expression vector resulted in similar findings : that OTF-2 protein enhanced DRA gene transcription , and that this effect requires an intact octamer element .

In vivo transient transfection studies utilizing an <dna>OTF-2 expression vector</dna> resulted in similar findings : that <protein>OTF-2 protein</protein> enhanced <dna>DRA gene</dna> transcription , and that this effect requires an intact <dna>octamer element</dna> .


Together these results constitute the first direct evidence of a positive role for the lymphoid-specific octamer-binding factor in DRA gene transcription .

Together these results constitute the first direct evidence of a positive role for the <protein>lymphoid-specific octamer-binding factor</protein> in DRA gene transcription .


Interferon-gamma potentiates the antiviral activity and the expression of interferon-stimulated genes induced by interferon-alpha in U937 cells .

<protein>Interferon-gamma</protein> potentiates the antiviral activity and the expression of <dna>interferon-stimulated genes</dna> induced by <cell_line>interferon-alpha in U937 cells</cell_line> .


Binding of type I interferon ( IFN-alpha/beta ) to specific receptors results in the rapid transcriptional activation , independent of protein synthesis , of IFN-alpha-stimulated genes ( ISGs ) in human fibroblasts and HeLa and Daudi cell lines .

Binding of <protein>type I interferon ( IFN-alpha/beta ) to specific receptors</protein> results in the rapid transcriptional activation , independent of protein synthesis , of <dna>IFN-alpha-stimulated genes</dna> ( <dna>ISGs</dna> ) in <cell_type>human fibroblasts</cell_type> and <cell_line>HeLa</cell_line> and <cell_line>Daudi cell lines</cell_line> .


The binding of ISGF3 ( IFN-stimulated gene factor 3 ) to the conserved IFN-stimulated response element ( ISRE ) results in transcriptional activation .

The binding of <protein>ISGF3</protein> ( <protein>IFN-stimulated gene factor 3</protein> ) to the conserved <dna>IFN-stimulated response element</dna> ( <dna>ISRE</dna> ) results in transcriptional activation .


This factor is composed of a DNA-binding protein ( ISGF3 gamma ) , which normally is present in the cytoplasm , and other IFN-alpha-activated proteins which preexist as latent cytoplasmic precursors ( ISGF3 alpha ) .

This factor is composed of a <protein>DNA-binding protein</protein> ( <protein>ISGF3 gamma</protein> ) , which normally is present in the cytoplasm , and other <protein>IFN-alpha-activated proteins</protein> which preexist as <protein>latent cytoplasmic precursors</protein> ( <protein>ISGF3 alpha</protein> ) .


We have found that ISG expression in the monocytic U937 cell line differs from most cell lines previously examined .

We have found that ISG expression in the <cell_line>monocytic U937 cell line</cell_line> differs from most <cell_line>cell lines</cell_line> previously examined .


U937 cells express both type I and type II IFN receptors , but only IFN-alpha is capable of inducing antiviral protection in these cells .

<cell_line>U937 cells</cell_line> express both <protein>type I and type II IFN receptors</protein> , but only <protein>IFN-alpha</protein> is capable of inducing antiviral protection in these cells .


Pretreatment with IFN-gamma potentiates the IFN-alpha -induced protection , but IFN-gamma alone does not have any antiviral activity .

Pretreatment with <protein>IFN-gamma</protein> potentiates the <protein>IFN-alpha</protein> -induced protection , but <protein>IFN-gamma</protein> alone does not have any antiviral activity .


ISG15 mRNA accumulation in U937 cells is not detectable before 6 h of IFN-alpha treatment , peaks at 24 h , and requires protein synthesis .

<rna>ISG15 mRNA</rna> accumulation in <cell_line>U937 cells</cell_line> is not detectable before 6 h of <protein>IFN-alpha</protein> treatment , peaks at 24 h , and requires protein synthesis .