Molecular Anatomy of RNA Polymerase

RNA 聚合酶的分子解剖学

• 批准号: 04454614
• 负责人: ISHIHAMA Akira
• 金额: $ 4.1万
• 依托单位: NATIONAL INSTITUTE OF GENETICS
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454614/
• 关键词: RNA polymerase; Transcription regulation; Transcription factor; Protein-protein contact; Structure-function relationship; Escherichia coli; Molecular anatomy; 酵母; 転写調節; 構造-機能相関; 蛋白-蛋白相互作用; 分子集合; 蛋白-DNA相互作用

Transcription is regulated by controlling the promoter selectivity of RNA polymerase. Bacterial RNA polymerase is composed of four major subunits, 2alpha, 1beta, 1beta' and 1sigma subunit. In order to understand how the promoter recognition properties of RNA polymerase is controlled by interaction with transcription factors, we carried out in this research the molecular anatomy of Escherichi coli RNA polymerase, in particular focussing on the contact site mapping on RNA polymerase subunits with various transcription factors. Results indicate that transcription factors can be classified based on the contact site on RNA polymerase. Class-I transcription factors were found to contact with the contact site I located at the C-terminal region of alpha subunit, while class-II factors make contact with the contact site II clustered in the C-terminal region of sigma subunit. Fine mapping revealed that the protein-protein communications between RNA polymerase and transcription factors involve narrow regions on both sides, each being composed of epitope size consisting of about 10 amino acid residues. In parallel, contact site mapping between RNA polymerase subunits has also been carried out.

通过控制RNA聚合酶的启动子选择性来调节转录。细菌RNA聚合酶由四个主要亚基，2alpha，1beta，1beta'和1 sigma亚基组成。为了了解RNA聚合酶的启动子识别特性如何通过与转录因子的相互作用来控制，我们在这项研究中进行了Escherichi Coli RNA聚合酶的分子解剖结构，特别是集中于与各种转录因子的RNA聚合酶亚基上的接触位点映射。结果表明，可以根据RNA聚合酶上的接触位点对转录因子进行分类。发现I类转录因子与位于Alpha亚基C末端区域的接触位点I接触，而II类因子使与Sigma亚基C末端区域的接触站点II接触。精细的映射显示，RNA聚合酶和转录因子之间的蛋白质 - 蛋白质通信涉及两侧的狭窄区域，每个区域由大约10个氨基酸残基组成的表位大小组成。同时，还进行了RNA聚合酶亚基之间的接触位点映射。

项目成果

Tanaka,K.,et al.: "Heterogeneity of the principal sigma factor in Escherichia coli:the rpoS gene product,38,is a principal sigma factor of RNA polymerase in stationary phase Escherichia coli." Proc.Natl.Acad.Sci.USA. 90. 3511-3515 (1993)

Tanaka, K., et al.：“大肠杆菌中主要 sigma 因子的异质性：rpoS 基因产物 38 是稳定期大肠杆菌中 RNA 聚合酶的主要 sigma 因子。”

Seong, B.L., Kobayashi, M., Nagata, K., Brownlee, G.G.and Ishihama, A.: "Comparison of two reconstitution systems for in vitro transcrition and replication of influenza virus." J.Biochem.111(4). 496-499 (1992)

Seong, B.L.、Kobayashi, M.、Nagata, K.、Brownlee, G.G. 和 Ishihama, A.：“流感病毒体外转录和复制的两种重建系统的比较。”

Azuma, Y., Yamagishi, M.and Ishihama, A.: "Subunits of the Schizosaccharomyces pombe RNA polymerase II : Enzyme purification and structure of the subunit 3 gene." Nucleic Acids Res.21. 3749-3754 (1993)

Azuma, Y.、Yamagishi, M. 和 Ishihama, A.：“裂殖酵母 RNA 聚合酶 II 的亚基：酶纯化和亚基 3 基因的结构。”

Zou,C.,et al.: "Mapping the Escherichia coli RNA polymerase contact site I for cAMP receptor protein." Mol.Microbiol.6. 2599-2605 (1992)

Zou,C.,et al.：“绘制 cAMP 受体蛋白的大肠杆菌 RNA 聚合酶接触位点 I”。

Ross,W.,et al.: "A third recognition element in prokaryotic promoters:Involvement of the α subunit of RNA polymerase." Science. 262. 1407-1413 (1993)

Ross, W., 等人：“原核启动子中的第三个识别元件：RNA 聚合酶 α 亚基的参与。科学”262. 1407-1413 (1993)