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通用转录因子RAP30与RNA聚合酶II结合，并阻止其非特异性地与DNA结合。

The general transcription factor RAP30 binds to RNA polymerase II and prevents it from binding nonspecifically to DNA.

作者信息

Killeen M T, Greenblatt J F

机构信息

Department of Molecular and Medical Genetics, University of Toronto, Ontario, Canada.

出版信息

Mol Cell Biol. 1992 Jan;12(1):30-7. doi: 10.1128/mcb.12.1.30-37.1992.

DOI:10.1128/mcb.12.1.30-37.1992
PMID:1729606
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC364066/
Abstract

RAP30/74 is a human general transcription factor that binds to RNA polymerase II and is required for initiation of transcription in vitro regardless of whether the promoter has a recognizable TATA box (Z. F. Burton, M. Killeen, M. Sopta, L. G. Ortolan, and J. F. Greenblatt, Mol. Cell. Biol. 8:1602-1613, 1988). Part of the amino acid sequence of RAP30, the small subunit of RAP30/74, has limited homology with part of Escherichia coli sigma 70 (M. Sopta, Z. F. Burton, and J. Greenblatt, Nature (London) 341:410-414, 1989). To determine which sigmalike activities of RAP30/74 could be attributed to RAP30, we purified human RAP30 and a RAP30-glutathione-S-transferase fusion protein that had been produced in E. coli. Bacterially produced RAP30 bound to RNA polymerase II in the absence of RAP74. Both partially purified natural RAP30/74 and recombinant RAP30 prevented RNA polymerase II from binding nonspecifically to DNA. In addition, nonspecific transcription by RNA polymerase II was greatly inhibited by RAP30-glutathione-S-transferase. DNA-bound RNA polymerase II could be removed from DNA by partially purified RAP30/74 but not by bacterially expressed RAP30. Thus, the ability of RAP30/74 to recruit RNA polymerase II to a promoter-bound preinitiation complex may be an indirect consequence of its ability to suppress nonspecific binding of RNA polymerase II to DNA.

摘要

RAP30/74是一种人类通用转录因子，它与RNA聚合酶II结合，并且在体外转录起始过程中是必需的，无论启动子是否具有可识别的TATA框（Z.F.伯顿、M.基林、M.索普塔、L.G.奥尔托兰和J.F.格林布拉特，《分子细胞生物学》8:1602 - 1613，1988年）。RAP30/74的小亚基RAP30的部分氨基酸序列与大肠杆菌σ70的部分序列具有有限的同源性（M.索普塔、Z.F.伯顿和J.格林布拉特，《自然》（伦敦）341:410 - 414，1989年）。为了确定RAP30/74的哪些类σ活性可归因于RAP30，我们纯化了人RAP30和在大肠杆菌中产生的RAP30 - 谷胱甘肽 - S - 转移酶融合蛋白。在没有RAP74的情况下，细菌产生的RAP30与RNA聚合酶II结合。部分纯化的天然RAP30/74和重组RAP30都能防止RNA聚合酶II非特异性地与DNA结合。此外，RAP30 - 谷胱甘肽 - S - 转移酶极大地抑制了RNA聚合酶II的非特异性转录。部分纯化的RAP30/74可以将与DNA结合的RNA聚合酶II从DNA上移除，但细菌表达的RAP30则不能。因此，RAP30/74将RNA聚合酶II募集到启动子结合的预起始复合物的能力可能是其抑制RNA聚合酶II与DNA非特异性结合能力的间接结果。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/364066/3c963f8e5ca2/molcellb00025-0057-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/364066/e742f2075297/molcellb00025-0054-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/364066/b78111bd2f58/molcellb00025-0055-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/364066/edd9454543e8/molcellb00025-0056-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/364066/a042ddb5f4d7/molcellb00025-0057-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/364066/3c963f8e5ca2/molcellb00025-0057-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/364066/e742f2075297/molcellb00025-0054-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/364066/b78111bd2f58/molcellb00025-0055-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/364066/edd9454543e8/molcellb00025-0056-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/364066/a042ddb5f4d7/molcellb00025-0057-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/364066/3c963f8e5ca2/molcellb00025-0057-b.jpg

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