SEQUENCE-SPECIFIC BINDING OF HUMAN ETS-1 TO THE T-CELL RECEPTOR ALPHA-GENE ENHANCER
SEQUENCE-SPECIFIC BINDING OF HUMAN ETS-1 TO THE T-CELL RECEPTOR ALPHA-GENE ENHANCER
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DOI：
10.1126/science.2237431
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发表时间：
1990-11-09
期刊：
影响因子：
56.9
通讯作者：
LEIDEN, JM
中科院分区：
文献类型：
--
作者：
HO, IC;BHAT, NK;LEIDEN, JM
人T细胞受体（TCR）α的表达基因由T细胞特异性转录增强子调节，该增强子位于C α 2基因片段的4.5个碱基（kb）3“处。核心增强子含有两个核蛋白结合位点，T α 1和T α 2，它们是完整增强子活性所必需的。T α 1含有共有环磷酸腺苷（cAMP）反应元件（CRE）并结合一组普遍表达的CRE结合蛋白。相反，与T α 2位点相互作用的转录因子尚未确定。在该报道中，使用了一种cDNA 11表达方案来分离编码T α 2结合蛋白表达的互补DNA（cDNA）。DNA序列分析表明，该克隆编码人ets-1原癌基因。用该cDNA克隆产生的溶原提取物含有β-特异性结合合成的T α 2寡核苷酸的半乳糖苷酶-Ets-1融合蛋白。Ets-1结合位点定位于T α 2的3“末端的17个碱基对（bp）区域。在该序列中5个核苷酸的突变消除了Ets-1结合和TCR α的活性。T细胞中的增强子。这些结果证明Ets-1以序列特异性方式结合人TCR α。增强子，并表明这种发育调节的原癌基因在调节TCR α中起作用。基因表达
Expression of the human T cell receptor (TCR) .alpha. gene is regulated by a T cell-specific transcriptional enhancer that is located 4.5 kilobases (kb) 3'' to the C.alpha.2, gene segement. The core enhancer contains two nuclear protein binding sites, T.alpha.1 and T.alpha.2, which are essential for full enhancer activity. T.alpha.1 contains a consensus cyclic adenosine monophosphate (cAMP) response element (CRE) and binds a set of ubiquitously expressed CRE binding proteins. In contrast, the transcription factors that interact with the T.alpha.2 site have not been defined. In this report, a .lambda.gt11 expression protocol was used to isolate a complementary DNA (cDNA) that programs the expression of a T.alpha.2 binding protein. DNA sequence analysis demonstrated that this clone encodes the human ets-1 proto-oncogene. Lysogen extracts produced with this cDNA clone contained a .beta.-galactosidase-Ets-1 fusion protein that bound specifically to a synthetic T.alpha.2 oligonucleotide. The Ets-1 binding site was localized to a 17-base pair (bp) region from the 3'' end of T.alpha.2. Mutation of five nucleotides within this sequence abolished both Ets-1 binding and the activity of the TCR .alpha. enhancer in T cells. These results demonstrate that Ets-1 binds in a sequence-specific fashion to the human TCR .alpha. enhancer and suggest that this developmentally regulated proto-oncogene functions in regulating TCR .alpha. gene expression.