RNA POLYMERASE SUBUNITS ALPHACTD AND SIGMA REGION4 IN COMPLEX WITH DNA

RNA 聚合酶亚基 ALPHACTD 和 Sigma Region4 与 DNA 复合

• 批准号: 8363408
• 负责人: CATHERINE L LAWSON
• 金额: $ 0.23万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363408
• 关键词: Bacteria; C-terminal; Complex; Crystallography; Cyclic AMP Receptor Protein; DNA; DNA-Directed RNA Polymerase; Data; Dimensions; Event; Funding; Genetic Transcription; Goals; Grant; Image; Light; Modeling; National Center for Research Resources; Organism; Principal Investigator; Process; Prokaryotic Cells; Protein Biosynthesis; RNA; Research; Research Infrastructure; Resolution; Resources; Source; Structure; Synchrotrons; Testing; Transcription Initiation; United States National Institutes of Health; Work; cost; promoter

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The transcription of DNA into RNA is a central process in the transfer of information required for protein synthesis. The study of transcription in prokaryotes provides a simple model for understanding transcription events in all living systems. The goal of this work is to characterize the structural factors involved in transcription initiation at Class I catabolite activator protein (CAP)-dependent promoters in bacteria. To this end, a crystal structure containing CAP, DNA and the C-Terminal Domain of the ¿¿ subunit of RNA polymerase (¿¿-CTD), which has been recently solved in this lab, is expanded to include the C-terminus of the ¿? subunit of RNA polymerase (¿? Region 4 or ¿?-R4). The ¿?-R4 of RNA polymerase is thought to make specific contacts with ¿¿-CTD and DNA in transcription initiation. Thus, a complex of CAP, DNA, ¿¿-CTD, and ¿?-R4 has been crystallized. Data has been collected on NSLS X25 to 3.5 ¿? resolution, but we have not yet been able to complete structure determination.We now have crystals that we believe consist of a subassembly of the CASD complex with alphaCTD, sigma region 4, and DNA (ASD). Several test images collected at the Cornell synchrotron indicate diffraction beyond 3 A, and lattice dimensions consistent with the proposed contents. If these crystals are indeed an ASD complex, determining this structure will be provide a very satisfactory means to complete several of the specific aims of this NIH-funded project.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 DNA 转录为 RNA 是蛋白质合成所需信息传递的核心过程。原核生物转录研究为理解所有生命系统中的转录事件提供了一个简单的模型。这项工作的目标是表征细菌中 I 类分解代谢物激活蛋白 (CAP) 依赖性启动子转录起始所涉及的结构因子。为此，本实验室最近解决了一个包含 CAP、DNA 和 RNA 聚合酶 (¿¿-CTD) 亚基 C 端结构域的晶体结构，该结构被扩展为包括 ¡ RNA 聚合酶亚基（??区域 4 或??-R4）。 RNA聚合酶的??-R4被认为在转录起始过程中与??-CTD和DNA发生特异性接触。因此，CAP、DNA、β-CTD和β-R4的复合物已结晶。数据已在 NSLS X25 至 3.5 ¿分辨率，但我们尚未能够完成结构确定。我们现在拥有晶体，我们认为该晶体由 CASD 复合物与 alphaCTD、sigma 区域 4 和 DNA (ASD) 的子组件组成。 在康奈尔同步加速器收集的几张测试图像表明衍射超过 3 A，并且晶格尺寸与建议的内容一致。 如果这些晶体确实是 ASD 复合物，那么确定这种结构将为完成该 NIH 资助项目的几个具体目标提供非常令人满意的方法。