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首页> 外文期刊>Nucleic Acids Research >ANALYSIS OF THE HUMAN TATA BINDING PROTEIN PROMOTER AND IDENTIFICATION OF AN ETS SITE CRITICAL FOR ACTIVITY
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ANALYSIS OF THE HUMAN TATA BINDING PROTEIN PROMOTER AND IDENTIFICATION OF AN ETS SITE CRITICAL FOR ACTIVITY

机译：人TATA结合蛋白启动子的分析及对ETS活性至关重要的位点的鉴定

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摘要

The TATA binding protein (TBP) is a general transcription factor required for initiation by all three eukaryotic nuclear RNA polymerases. Little is known about how TBP gene expression is regulated. To identify sequence elements and proteins contributing to human TBP (hTBP) gene transcription, we have characterized the promoter in two human cell lines. Multiple 5'-ends of TBP mRNA mapped throughout a 111 bp region immediately upstream of a previously reported hTBP cDNA. Upon transient transfection into cells, the hTBP 5'-flanking region was shown to contain a fairly active promoter. The cis-acting elements responsible for this promoter activity in Namalwa and HeLa cells were localized in vivo by deletion analysis. The minimal promoter defined from these experiments was a 54 bp region that encompassed all but one minor start site and contained a functional Ets protein consensus binding site, which was shown to be required for promoter activity in both cell lines. The importance of other potential elements to promoter activity was found to differ between the two cell lines. Consistent with that finding, different complexes were formed on promoter-containing DNA fragments upon incubation with nuclear extracts prepared from the different cells.
机译：TATA结合蛋白（TBP）是由所有三种真核核RNA聚合酶起始所需的一般转录因子。关于TBP基因表达如何调控知之甚少。为了鉴定有助于人类TBP（hTBP）基因转录的序列元件和蛋白质，我们已经鉴定了两种人类细胞系中的启动子。 TBP mRNA的多个5'端位于先前报道的hTBP cDNA上游111 bp区域。在瞬时转染到细胞中后，hTBP 5'-侧翼区域显示含有相当活跃的启动子。通过缺失分析，将负责Namalwa和HeLa细胞中该启动子活性的顺式作用元件定位在体内。从这些实验中定义的最小启动子是一个54 bp的区域，除了一个较小的起始位点外，其余全部都包含在内，并包含功能性Ets蛋白共有结合位点，这在两个细胞系中均显示为启动子活性所必需。发现其他潜在元件对启动子活性的重要性在两种细胞系之间有所不同。与该发现一致的是，与由不同细胞制备的核提取物一起孵育后，在含启动子的DNA片段上形成了不同的复合物。

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