CLONING AND PROPERTIES OF THE CAENORHABDITIS-ELEGANS TATA-BOX-BINDING PROTEIN
CLONING AND PROPERTIES OF THE CAENORHABDITIS-ELEGANS TATA-BOX-BINDING PROTEIN
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DOI：
10.1073/pnas.90.20.9673
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发表时间：
1993-10-15
影响因子：
11.1
通讯作者：
TJIAN, R
中科院分区：
文献类型：
--
作者：
LICHTSTEINER, S;TJIAN, R
秀丽隐杆线虫已成为在遗传水平上研究发育过程的首选生物。我们已经着手建立一个体外系统来研究C.优美的作为第一步，我们在这里报告的cDNA编码的克隆C。线虫TATA盒结合蛋白（CeTBP）。我们使用“降落PCR”产生一个特定的DNA探针，该探针来自迄今为止克隆的所有TBP基因中保守的C-末端区域。从噬菌体λ cDNA文库中分离出几个编码延伸开放阅读框的克隆。从cDNA推导的CeTBP的完整氨基酸序列揭示了一个37 kDa的蛋白质，其C-末端区域与迄今为止测序的所有其他TBP cDNA具有扩展的序列相似性。然而，CeTBP的N-末端区域（氨基酸1-153）不显示与来自其它生物体的TBP的任何同源性。有趣的是，该分子的N-末端部分含有三个短的直接重复序列。纯化的重组CeTBP与TATA盒序列特异性结合，与转录因子TFIIA和TFIIB相互作用，并且当在HeLa和C中通过体外转录测定时能够替代MID基础活性。线虫核提取物。因此，CeTBP是基础转录因子。
The nematode Caenorhabditis elegans has become an organism of choice for the study of developmental processes at the genetic level. We have undertaken to develop an in vitro system to study transcription in C. elegans. As a first step we report here the cloning of the cDNA encoding the C. elegans TATA-box-binding protein (CeTBP). We used ''touchdown PCR'' to generate a specific DNA probe derived from the C-terminal region conserved in all TBP genes cloned to date. Several clones encoding an extended open reading frame were isolated from a phage lambda cDNA library. The complete amino acid sequence of CeTBP deduced from the cDNA reveals a protein of 37 kDa with an extended sequence similarity in the C-terminal region with all other TBP cDNAs sequenced so far. The N-terminal region of CeTBP (amino acids 1-153), however, does not show any homology with TBPs from other organisms. Interestingly, the N-terminal portion of the molecule contains three short direct repeats. Purified recombinant CeTBP binds specifically to the TATA box sequence, interacts with transcription factors TFIIA and TFIIB, and is able to substitute for the MID basal activity when assayed by in vitro transcription in both HeLa and C. elegans nuclear extracts. CeTBP is therefore a basal transcription factor.