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RNA聚合酶III介导的TATA框体外转录。聚合酶III型复合物中TATA结合蛋白的证据。

TATA box-mediated in vitro transcription by RNA polymerase III. Evidence for TATA-binding protein in a polymerase III type complex.

作者信息

Mitchell M T, Benfield P A

机构信息

Du Pont Merck Pharmaceutical Co., Wilmington, Delaware 19880-0328.

出版信息

J Biol Chem. 1993 Jan 15;268(2):1141-50.

PMID:7678250
Abstract

We have shown previously that under specific conditions, a TATA box will mediate efficient in vitro transcription by RNA polymerase (pol) III in the absence of a PSE or other promoter elements. The reaction requires a HeLa cell phosphocellulose protein fraction, fraction B, which must be preincubated with the template DNA. Fraction B does not contain any detectable pol II type transcription factor IID (TFIID) activity. In this report, the relationship between fraction B and TFIID was further examined. Purified human TATA-binding protein (TBP) can substitute for fraction B to mediate TATA-dependent pol III transcription. Both TBP and fraction B prefer a reverse TATA box for pol III transcription, yet TBP bound to a reverse TATA box functions poorly for pol II transcription. Like TFIID, fraction B forms a template-committed complex with TATA-containing promoters. TBP, however, will not template commit for pol III transcription unless premixed with phosphocellulose fraction C. TBP-mediated pol III transcription is also more sensitive to the detergent Sarkosyl (N-lauroylsarcosine, Sigma) than is the fraction B reaction unless it is premixed with fraction C. Together, the data suggest that TBP can complex with a component of fraction C, and this complex is then functionally equivalent to fraction B. We propose that fraction B contains TBP in a complex with some other component(s) of the pol III transcription machinery and that this B complex TBP may be specific for pol III transcription.

摘要

我们之前已经表明，在特定条件下，TATA框在没有近端序列元件（PSE）或其他启动子元件的情况下，将介导RNA聚合酶（pol）III高效的体外转录。该反应需要HeLa细胞磷酸纤维素蛋白组分B，它必须与模板DNA预孵育。组分B不含有任何可检测到的pol II型转录因子IID（TFIID）活性。在本报告中，进一步研究了组分B与TFIID之间的关系。纯化的人TATA结合蛋白（TBP）可以替代组分B来介导依赖TATA的pol III转录。TBP和组分B在pol III转录中都更喜欢反向TATA框，然而，与反向TATA框结合的TBP在pol II转录中功能较差。与TFIID一样，组分B与含TATA的启动子形成模板结合复合物。然而，除非与磷酸纤维素组分C预混合，TBP不会为pol III转录进行模板结合。TBP介导的pol III转录对去污剂十二烷基肌氨酸钠（N-月桂酰肌氨酸，Sigma）也比组分B反应更敏感，除非它与组分C预混合。总之，数据表明TBP可以与组分C的一种成分形成复合物，并且这种复合物在功能上等同于组分B。我们提出组分B包含与pol III转录机制的一些其他成分形成复合物的TBP，并且这种B复合物TBP可能对pol III转录具有特异性。

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J Biol Chem. 1993 Jan 15;268(2):1141-50.
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引用本文的文献

1
Role of TATA box sequence and orientation in determining RNA polymerase II/III transcription specificity.TATA 框序列和方向在决定 RNA 聚合酶 II/III 转录特异性中的作用。
Nucleic Acids Res. 1996 Aug 1;24(15):3100-6. doi: 10.1093/nar/24.15.3100.
2
Gene expression: surprises from the class III side.基因表达：来自III类的惊喜。
Mol Cell Biochem. 1993 Jul 7;124(1):85-9. doi: 10.1007/BF01096385.
3
RNA polymerase II/III transcription specificity determined by TATA box orientation.由TATA框方向决定的RNA聚合酶II/III转录特异性。
Proc Natl Acad Sci U S A. 1995 Sep 12;92(19):8606-10. doi: 10.1073/pnas.92.19.8606.