STRUCTURAL STUDIES OF RNA POLYMERASE COMPLEXES

RNA聚合酶复合物的结构研究

• 批准号: 6525689
• 负责人: ELIZABETH A CAMPBELL
• 金额: $ 4.62万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6525689
• 关键词: DNA directed RNA polymerase; X ray crystallography; adenosine triphosphate; antibiotics; bacterial proteins; dinucleotide; enzyme complex; enzyme structure; genetic transcription; molecular cloning; rifamycins; structural biology; transcription factor

RNA polymerase is the central enzyme of gene expression and regulation in bacteria. Recently this laboratory has solved the X-ray crystal structure of core RNAP (the processive form of the enzyme) opening a path for subsequent structural studies of this enzyme's interaction with additional subunits and ligands. The overall goal of this proposal is to understand the mechanism of transcription in relation to the structure of the enzyme. Revelation of the holoenzyme structure would add great insight to how the sigma factor allows RNAP to acquire promoter specificity, initiate transcription and regulate gene inhibitors of the enzyme, will increase our current understanding of the transcription mechanisms. These structures are also extremely relevant to medical research for anti-biotic design against bacterial pathogens. This proposal will be accomplished by X-ray crystallography of core RNAP complexed with the following factors: 1) the housekeeping sigma factor (SigA); 2) an initiating nucleotide tri- phosphate, (ATP); 3) a dinucleotide (ApU) and 4) the antibiotics rifampican and streptolydigin.

RNA聚合酶是细菌基因表达和调节的核心酶。最近，该实验室解决了核心 RNAP（酶的加工形式）的 X 射线晶体结构，为后续对该酶与其他亚基和配体相互作用的结构研究开辟了道路。该提案的总体目标是了解与酶结构相关的转录机制。全酶结构的揭示将有助于深入了解 sigma 因子如何使 RNAP 获得启动子特异性、启动转录并调节酶的基因抑制剂，将增加我们目前对转录机制的理解。这些结构也与针对细菌病原体的抗生素设计的医学研究极为相关。该提议将通过核心 RNAP 与以下因子复合的 X 射线晶体学来完成：1) 管家西格玛因子 (SigA)； 2) 起始核苷酸三磷酸（ATP）； 3) 二核苷酸 (ApU) 和 4) 抗生素利福平和链球菌素。