THE N-TERMINAL DOMAIN OF THE HUMAN TATA-BINDING PROTEIN PLAYS A ROLE IN TRANSCRIPTION FROM TATA-CONTAINING RNA-POLYMERASE-II AND RNA-POLYMERASE-III PROMOTERS
THE N-TERMINAL DOMAIN OF THE HUMAN TATA-BINDING PROTEIN PLAYS A ROLE IN TRANSCRIPTION FROM TATA-CONTAINING RNA-POLYMERASE-II AND RNA-POLYMERASE-III PROMOTERS
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DOI：
10.1002/j.1460-2075.1994.tb06366.x
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发表时间：
1994-03-01
期刊：
影响因子：
11.4
通讯作者：
TORA, L
中科院分区：
文献类型：
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作者：
LESCURE, A;LUTZ, Y;TORA, L
在真核生物中，TATA盒结合蛋白（TBP）是所有三类核RNA聚合酶的转录起始复合物的组成部分。在这项研究中，我们已经调查了从RNA聚合酶（Pol）I，II和III启动子的转录起始的人TBP的N-末端区域的作用，通过使用三种单克隆抗体（mAb）。每种抗体识别人TBP的N-末端结构域中的不同表位。我们证明，这些抗体差异影响转录不同类别的启动子。一种抗体mAb 1C 2和包含其表位的合成肽选择性地抑制来自含TATA启动子的体外转录，但不抑制来自无TATA启动子的体外转录，无论它们是由Pol II还是Pol m转录。另一方面，Pol I的转录没有受到影响。另外两种抗体和它们各自的表位肽不影响从任何测试的启动子的转录。添加顺序的实验表明，mAb 1C 2没有阻止TBP与TATA盒的结合或TBP-TFIIA-TFIIB复合物的形成，而是抑制了随后的预起始复合物形成步骤。这些数据表明，一个确定的区域内的N-末端结构域的人TBP可能参与特定的蛋白质-蛋白质的相互作用所需的组装功能preinitiation复合物的TATA-含有，但不TATA-少启动子。
In eukaryotes, the TATA box binding protein (TBP) is an integral component of the transcription initiation complexes of all three classes of nuclear RNA polymerases. In this study we have investigated the role of the N-terminal region of human TBP in transcription initiation from RNA polymerase (Pol) I, II and III promoters by using three monoclonal antibodies (mAbs). Each antibody recognizes a distinct epitope in the N-terminal domain of human TBP. We demonstrate that these antibodies differentially affect transcription from distinct classes of promoters. One antibody, mAb1C2, and a synthetic peptide comprising its epitope selectively inhibited in vitro transcription from TATA-containing, but not from TATA-less promoters, irrespective of whether they were transcribed by Pol II or Pol m. Transcription by Pol I, on the other hand, was not affected. Two other antibodies and their respective epitope peptides did not affect transcription from any of the promoters tested. Order of addition experiments indicate that mAb1C2 did not prevent binding of TBP to the TATA box or the formation of the TBP-TFIIA-TFIIB complex but rather inhibited a subsequent step of preinitiation complex formation. These data suggest that a defined region within the N-terminal domain of human TBP may be involved in specific protein- protein interactions required for the assembly of functional preinitiation complexes on TATA-containing, but not on TATA-less promoters.