The same target sequences are differentially important for activation of the interleukin 2 receptor alpha-chain gene in two distinct T-cell lines.

Toledano, Michel B.; Roman, D G; Halden, N F; Lin, B B; Leonard, Warren J.

doi:10.1073/pnas.87.5.1830

Cited by 37 publications

(25 citation statements)

References 35 publications

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“…Though we did not detect cooperativity in binding in vitro, one model is that in vivo, SRF is capable of recruiting NF-cB to its adjacent binding site. The importance of this mechanism for the regulation of the IL-2Rt gene is supported by previous studies in which mutations in the sequences that bind SRF severely impair IL-2Ra promoter function in T cells (3,59). However, these previous studies do not address the contribution of SRF to T-cell specificity in the IL-2R gene, because the promoter mutants and levels of binding activity were not examined in non-T cells.…”

Section: Discussionsupporting

confidence: 57%

“…Sequences within the ILT oligonucleotide resembled a motif called CArG, [CC(A/T)6GG], which binds a nuclear regulatory protein known as SRF (15,32,33,36,41,42,60). Others have previously noted similarities between a protein complex with the IL-2R promoter and SRF (3,38,59). We therefore compared the ILT complex with SRF by binding to a CArG box-containing oligonucleotide derived from Xenopus -y-actin gene sequences (SRE, a gift of R. Prywes).…”

Section: Methodsmentioning

confidence: 99%

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Interaction between NF-kappa B- and serum response factor-binding elements activates an interleukin-2 receptor alpha-chain enhancer specifically in T lymphocytes.

et al. 1993

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We find that a short enhancer element containing the NF-KB binding site from the interleukin-2 receptor a-chain gene (IL-2Ra) is preferentially activated in T cells. The IL-2Rca enhancer binds NF-KB poorly and is only weakly activated by the NF-KB site alone. Serum response factor (SRF) binds to a site adjacent to the NF-KB site in the IL-2R enhancer, and both sites together have strong transcriptional activity specifically in T cells. Surprisingly, the levels of SRF constitutively expressed in T cells are consistently higher than in other cell types. Overexpression of SRF in B cells causes the IL-2R enhancer to function as well as it does in T cells, suggesting that the high level of SRF binding in T cells is functionally important.The a-chain of the interleukin-2 receptor (IL-2Ra) is a key surface protein that is expressed during the activation of T lymphocytes. Interleukin-2 (IL-2) is a major regulator of T-cell proliferation and death (23,53,54). The IL-2Rao protein is a component of the high-affinity receptor for IL-2 (62). The IL-2 receptor is expressed early in mouse T-cell differentiation on CD4-CD8-thymocytes, in which it may play a role in the expansion of thymocyte number (58). The IL-2Ra protein, and presumably its gene, are turned off at later stages of thymic development. In mature T cells, the IL-2Ra gene can be reexpressed following antigen stimulation. This is a tightly regulated process-resting T lymphocytes express virtually no IL-2Ra, whereas antigen-activated T cells have surface expression of 3 x 104 to 6 x 104 IL-2Rot molecules (62). The IL-2Ra gene is most commonly expressed in T cells but can be expressed also in other hematopoietic cells (62). During antigen activation of mature T cells, the promoter of the IL-2Rao gene is regulated by a DNA sequence that binds the NF-KB protein complex (1,3, 4,8,27,44).NF-KB is a protein complex that enables a large number of genes to be rapidly induced in response to extracellular stimuli (16, 24). Two polypeptides, termed p5O and p65, together form the NF-KB transcriptional transactivator by specifically binding to a 10-nucleotide DNA sequence (2, 50). The recent cloning of the subunits of NF-KB reveals that they belong to the Rel family of DNA-binding protein genes, which includes p5O, p49/p5OB (lyt-10), p65, c-rel, RelB,7,12,21,34,35,45,49,50,63). Mature B cells express high constitutive levels of NF-KB (pSO/p65 heterodimer) binding activity in the nucleus (50). Most other

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Section: Discussionsupporting

confidence: 57%

Section: Methodsmentioning

confidence: 99%

Interaction between NF-kappa B- and serum response factor-binding elements activates an interleukin-2 receptor alpha-chain enhancer specifically in T lymphocytes.

et al. 1993

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“…For example, the IL-2Ro promoter contains a KB site that is important for IL-2Rox expression in human T-lymphotrophic virus type I-transformed MT-2 T cells but not Jurkat T cells, whereas the Ig KB site is active in both cell types (13,47 (25). A dimerization domain has been localized much closer to the C terminus within the Rel homology region (28), remote from the region that we have characterized.…”

Section: Discussionmentioning

confidence: 99%

“…distinct and why single sites can manifest different activities in distinct cell types (13,28,47). For example, the IL-2Ro promoter contains a KB site that is important for IL-2Rox expression in human T-lymphotrophic virus type I-transformed MT-2 T cells but not Jurkat T cells, whereas the Ig KB site is active in both cell types (13,47 (25).…”

Section: Discussionmentioning

confidence: 99%

N-terminal DNA-binding domains contribute to differential DNA-binding specificities of NF-kappa B p50 and p65.

Toledano¹,

Ghosh²,

Trinh³

1993

Mol. Cell. Biol.

Self Cite

118

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We previously reported that either oxidation or alkylation of NF-cB in vitro abrogates DNA binding. We used this phenomenon to help elucidate structural determinants of NF-KB binding. We now demonstrate that Cys-62 of NF-KB p50 mediates the redox effect and lies within an N-terminal region required for DNA binding but not for dimerization. Several (16,23,33,40). All of these proteins are members of the Rel family of proteins, also denoted the NRD family, for NF-KB/Rel/dorsal. More recently recognized members of this family are RelB/I-Rel (41, 42) and p49 (also denoted p5OB), which is similar to p50 in that both are derived from larger precursor proteins and can interact functionally with p65 (10, 43).In unstimulated cells, NF-KB is present in the cytoplasm in a latent form consisting of p50 (or p49) and p65 and bound via p65 to an inhibitory molecule, IKB (4-6, 18, 22). Cellular activation results in dissociation of NF-KB from IKB and its translocation to the nucleus as an active DNA-binding complex (4, 5). In vitro, p50, p65, and c-Rel can bind DNA as homodimers or as pSO-p65 or p50-c-Rel heterodimers (8,23,33,49 Fig. 1, lane 1), a p50 construct corresponding to amino acids 1 to 399 (HindIII-RsaI fragment) (P501_399) was used. The binding properties of P501-399 and P501-502 are indistinguishable (23; data not shown). p65 constructs were derived from a BamHI fragment spanning the entire human cDNA coding sequence (40). Single point mutation, N-terminal deletion, and pSONTmyc constructs were prepared by site-directed mutagenesis as previously described (47) with a Bio-Rad kit. Following subcloning into pBluescript SK(-) (Stratagene), mutant constructs were resequenced. Chimeras between p50 and p65 were prepared by a two-step polymerase chain reaction method (19), and correctness was confirmed by sequencing the N-terminal regions extending through the point of "fusion" between p50 and p65.In vitro transcription and translation. In vitro transcription was carried out with constructs in which cDNAs were subcloned in pBluescript SK(-) and the mRNA cap kit (Stratagene). p50 and chimeric constructs were linearized with XbaI and transcribed with T7 RNA polymerase. p65 constructs were linearized with HindIII and transcribed with T3 RNA polymerase. Wheat germ extracts were used for the experiments shown in Fig. 1. Reticulocyte lysates were used for the experiments shown in Fig. 3 and 4. The data summarized in Fig. 2

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“…As illustrated in Figure 5, the human IL-2Ra promoter contains three well-characterized positive regulatory regions (PRRs). PRRI is located at 7299 to 7228 relative to the major transcription initiation site and binds to NF-kB serum response factor (SRF) and Sp1 (Ballard et al, 1989;Bohnlein et al, 1988;Cross et al, 1989;Leung and Nabel, 1988;Roman et al, 1990;Toledano et al, 1990). PRRII spans the region between 7137 to 764 and binds to an Ets family protein, Elf-1, and the high mobility group proteins, HMG-I/Y Lecine et al, 1996).…”

Section: How Do Stat5 Proteins Regulate Transcription Of Target Genes?mentioning

confidence: 99%

The role of Stat5a and Stat5b in signaling by IL-2 family cytokines

2000

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The same target sequences are differentially important for activation of the interleukin 2 receptor alpha-chain gene in two distinct T-cell lines.

Cited by 37 publications

References 35 publications

Interaction between NF-kappa B- and serum response factor-binding elements activates an interleukin-2 receptor alpha-chain enhancer specifically in T lymphocytes.

Interaction between NF-kappa B- and serum response factor-binding elements activates an interleukin-2 receptor alpha-chain enhancer specifically in T lymphocytes.

N-terminal DNA-binding domains contribute to differential DNA-binding specificities of NF-kappa B p50 and p65.

The role of Stat5a and Stat5b in signaling by IL-2 family cytokines

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