<i>In vivo</i>footprinting of the human IL-2 gene reveals a nuclear factor bound to the transcription start site in T cells

Brunvand, Mark W.; Krumm, Anton; Groudine, Mark

doi:10.1093/nar/21.20.4824

Cited by 31 publications

(28 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The location of the in vitro-positioned nucleosome (Ϫ60 to Ϫ210) lies approximately within the bleomycin-footprinted region suggesting that the bleomycin footprint may in part represent a positioned nucleosome. It has previously been shown that a constitutive DNase I footprint occurs across the TATA box region of the IL-2 promoter in Jurkat T cells and this has been interpreted as representing a paused polymerase complex (40,41). Such a complex together with a FIGURE 7.…”

Section: Discussionmentioning

confidence: 99%

“…These experiments revealed that in resting mouse and human T cells, when the IL-2 gene is silent, the proximal promoter region is inaccessible to DNase I and restriction enzyme digestion. Following simulation, new DNase I hypersensitive sites appear suggesting changes in chromatin structure (37)(38)(39)(40)(41).…”

Section: T He Generation Of T Cell-mediated Immune Responses Ismentioning

confidence: 99%

“…Histones were reconstituted onto radiolabeled IL-2 promoter fragments by high-salt to low-salt step-wise exchange with 30-to 60-fold excess of either isolated chicken erythrocyte nucleosome core particles or H1-depleted oligonucleosomes as previously described (41,45). The in vitro-reconstituted nucleosomes were routinely purified from both contaminating free DNA and unbound proteins by loading onto a pre-equilibrated 5-20% 10 ml sucrose gradient in buffer containing 1 M Tris, pH 7.5, 500 mM EDTA, and 100 mM PMSF and centrifuged at 4°C for 16 h at 35,000 rpm.…”

Section: Chromatin Reconstitution and Characterization Of Nucleosome mentioning

confidence: 99%

See 2 more Smart Citations

The Human IL-2 Gene Promoter Can Assemble a Positioned Nucleosome That Becomes Remodeled Upon T Cell Activation

Attema

Reeves

Murray

et al. 2002

The Journal of Immunology

View full text Add to dashboard Cite

Controlled production of the cytokine IL-2 plays a key role in the mammalian immune system. Expression from the gene is tightly regulated with no detectable expression in resting T cells and a strong induction following T cell activation. The IL-2 proximal promoter (+1 to −300) contains many well-defined transcriptional activation elements that respond to T cell stimulation. To determine the role of chromatin structure in the regulation of interleukin-2 gene transcription, nucleosome assembly across the IL-2 promoter region was examined using in vitro chromatin reconstitution assays. The IL-2 promoter assembles a nucleosome that is both translationally and rotationally positioned, spanning some of the major functional control elements. The binding of transcription factors to these elements, with the exception of the architectural protein HMGA1, was occluded by the presence of the nucleosome. Analysis of the chromatin architecture of the IL-2 gene in Jurkat T cells provided evidence for the presence of a similarly positioned nucleosome in vivo. The region encompassed by this nucleosome becomes remodeled following activation of Jurkat T cells. These observations suggest that the presence of a positioned nucleosome across the IL-2 proximal promoter may play an important role in maintaining an inactive gene in resting T cells and that remodeling of this nucleosome is important for gene activation.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: T He Generation Of T Cell-mediated Immune Responses Ismentioning

confidence: 99%

Section: Chromatin Reconstitution and Characterization Of Nucleosome mentioning

confidence: 99%

See 1 more Smart Citation

The Human IL-2 Gene Promoter Can Assemble a Positioned Nucleosome That Becomes Remodeled Upon T Cell Activation

Attema

Reeves

Murray

et al. 2002

The Journal of Immunology

View full text Add to dashboard Cite

show abstract

“…The highly inducible nature of the IL-2 promoter seems to be the result of coordinate binding of many transcription factors to their recognition sequences on the promoter leading to the assembly of a functional unit (3,4,11,14,(21)(22)(23)(24). Such a unit has been termed an enhanceosome on the IFN-␤ promoter (25).…”

mentioning

confidence: 99%

The Role of High-Mobility Group I(Y) Proteins in Expression of IL-2 and T Cell Proliferation

Himes

Reeves

Attema

et al. 2000

The Journal of Immunology

View full text Add to dashboard Cite

The high-mobility group I(Y) (HMGI(Y)) family of proteins plays an important architectural role in chromatin and have been implicated in the control of inducible gene expression. We have previously shown that expression of HMGI antisense RNA in Jurkat T cells inhibits the activity of the IL-2 promoter. Here we have investigated the role of HMGI(Y) in controlling IL-2 promoter-reporter constructs as well as the endogenous IL-2 gene in both Jurkat T cells and human PBL. We found that the IL-2 promoter has numerous binding sites for HMGI(Y), which overlap or are adjacent to the known transcription factor binding sites. HMGI(Y) modulates binding to the IL-2 promoter of at least three transcription factor families, AP-1, NF-AT and NF-κB. By using a mutant HMGI that cannot bind to DNA but can still interact with the transcription factors, we found that DNA binding by HMGI was not essential for the promotion of transcription factor binding. However, the non-DNA binding mutant acts as a dominant negative protein in transfection assays, suggesting that the formation of functional HMGI(Y)-containing complexes requires DNA binding as well as protein:protein interactions. The alteration of HMGI(Y) levels affects IL-2 promoter activity not only in Jurkat T cells but also in PBL. Importantly, we also show here that expression of the endogenous IL-2 gene as well as proliferation of PBL are affected by changes in HMGI(Y) levels. These results demonstrate a major role for HMGI(Y) in IL-2 expression and hence T cell proliferation.

show abstract

“…It belongs to the Rel family of transcriptional regulatory proteins (26,31). The p50 subunit is specifically bound to an intron enhancer site in a number of eukaryotic genes (33)(34)(35)(36). NF-B is involved in cross-talk with other transcription regulators.…”

Section: Cholecystokinin (Cck)mentioning

confidence: 99%

Identification of Overlapping AP-2/NF-κB-responsive Elements on the Rat Cholecystokinin Gene Promoter

Katsel

Greenstein²

2001

Journal of Biological Chemistry

View full text Add to dashboard Cite

In this study we evaluate both proximal and more distal transcriptional regulation of the 5 flanking region of the rat cholecystokinin gene in transfected GH3 (rat pituitary tumor) cells. Transcriptional activity was measured on the intact (؊400 to ؉73) 5 flanking region of cholecystokinin (CCK), as well as with DNA constructs, which were deleted in both the conventional 5 to 3, as well as an unconventional 3 to 5 direction. Our in vivo studies indicate complex phorbol ester and forskolin interactions in the 10-base pair region between ؊130 and ؊140. We conclude, there are at least two transcriptional factors involved in regulation of the rat CCK transcription in this region. In vitro studies utilizing heterologous nuclear (HeLa) extract, as well as purified transcription factors AP-2 and NF-B, identify overlapped AP-2-and NF-B-responsive elements within the 17-base pair sequence between ؊149 and ؊134 of the distal 5 flanking region. In this region complex transcriptional regulation occurs, which indicates inhibition of AP-2 CCK promoter complexing by NF-B. Sixpoint mutations introduced into this sequence prevent AP-2 and NF-B binding to CCK promoter, as well as its transcriptional activation by phorbol ester and forskolin in GH3 cells. Cholecystokinin (CCK),1 a prototypical brain-gut peptide (1-5), has hemacrine (3, 6) and autocrine (7) action in the gut. In the brain CCK is a neurotransmitter (8, 9). There is a commonly accepted gut, as well as brain, CCK transcription initiation site in the rat (10), mouse (11), and man (12). Alternatively, more distal 5Ј CCK transcription start sites have been identified in the brain of the rat (13). Transcriptional control of CCK is regulated by multiple factors. These include phorbol ester (14 -16), cAMP (14,(17)(18)(19)(20), and gonadal steroids (21), as well as glucocorticoids (22). Several cis-elements, important in transcriptional control, have been identified within the first 119 bp of the 5Ј flanking region of the rat CCK gene (10). This 119-bp fragment contains a sequence homologous with a 12-Otetradecanoylphorbol 13-acetate (TPA)-responsive element of the c-fos gene, as well as a cAMP-responsive element identified in the proenkephalin gene (23). This sequence binds several transcription factors including AP-1, jun, and fos homodimers, as well as cAMP-responsive element-binding protein (16,19). Two other recognition sites on the human CCK gene are Sp1 (at Ϫ39 to Ϫ34) and E-box or upstream stimulatory factor (at Ϫ97 to Ϫ92) (16). Additionally, a negative interaction was detected between TPA-responsive element and E-box or upstream stimulatory factor-responsive element (24).TPA and forskolin activate specific nuclear proteins, which regulate gene transcription. These proteins include AP-2 (25) and NF-B (26, 27). Transcription factor AP-2 is activated either directly or indirectly by two signal transduction pathways (25). One route involves cAMP-dependent protein kinase A. The other involves phorbol ester and diacylglycerol-activated protein kinase C (28). AP-2 specifi...

show abstract

In vivofootprinting of the human IL-2 gene reveals a nuclear factor bound to the transcription start site in T cells

Cited by 31 publications

References 32 publications

The Human IL-2 Gene Promoter Can Assemble a Positioned Nucleosome That Becomes Remodeled Upon T Cell Activation

The Human IL-2 Gene Promoter Can Assemble a Positioned Nucleosome That Becomes Remodeled Upon T Cell Activation

The Role of High-Mobility Group I(Y) Proteins in Expression of IL-2 and T Cell Proliferation

Identification of Overlapping AP-2/NF-κB-responsive Elements on the Rat Cholecystokinin Gene Promoter

Contact Info

Product

Resources

About