STRUCT STUDY OF BACTERIOPHAGE N4 RNA POLYMERASE TRANSCRIPTION INITIATION COMPLEX

噬菌体N4 RNA聚合酶转录起始复合物的结构研究

• 批准号: 8363539
• 负责人: Katsuhiko Murakami
• 金额: $ 1.95万
• 依托单位: CORNELL UNIVERSITY
• 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/8363539
• 关键词: Bacteriophage N4; Biological Models; Complex; Crystallization; DNA; DNA-Directed RNA Polymerase; Data Set; Funding; Genetic Transcription; Grant; Home environment; L-Selenomethionine; Macromolecular Complexes; Methods; Molecular; Molecular Structure; National Center for Research Resources; Principal Investigator; Proteins; Reaction; Research; Research Infrastructure; Resolution; Resources; Roentgen Rays; Source; Staging; Structure; Time; Transcription Initiation; United States National Institutes of Health; beamline; cost; enzyme structure

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. Structural study of bacteriophage N4 RNA polymerase transcription initiation complex We are using bacteriophage N4 RNA polymerase (N4 RNAP) as a model system to understand transcription mechanism. We have solved apo enzyme structure at 2.3A resolution by using SeMet-protein MAD method and dataset obtained at X25/NSLS. Recently, we have solved N4 RNAP-DNA complex structure by molecular replacement. The dataset was collected with home source X-ray and the resolution is 2.4A. Nobody solved the structure of RNAP-DNA-substrate complex at transcription initiation stage (transcription initiation complex). We prepared initiation complex crystal by co-crystallization and found it diffracts at 2.4A resolution with home source X-ray. To understand molecular detail of transcription imitation reaction, we want to request beamline time of F1/CHESS to collect transcription initiation complex crystal dataset at atomic resolution. The structure will be the first snap shot of transcription initiation stage and it tells us why RNAP is able to carry out de novo reaction. This project is funded by NIH.

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。 噬菌体 N4 RNA 聚合酶转录起始复合物的结构研究 我们使用噬菌体 N4 RNA 聚合酶 (N4 RNAP) 作为模型系统来了解转录机制。我们通过使用 SeMet-蛋白质 MAD 方法和在 X25/NSLS 获得的数据集以 2.3A 分辨率解析了 apo 酶结构。最近，我们通过分子置换解析了N4 RNAP-DNA复合物结构。数据集采用家用X射线采集，分辨率为2.4A。没有人解析转录起始阶段RNAP-DNA-底物复合物（转录起始复合物）的结构。我们通过共结晶制备了引发复合晶体，并发现它在家用X射线下的衍射分辨率为2.4A。为了了解转录模仿反应的分子细节，我们希望请求 F1/CHESS 的光束线时间来收集原子分辨率的转录起始复杂晶体数据集。该结构将是转录起始阶段的第一个快照，它告诉我们为什么RNAP能够进行从头反应。该项目由美国国立卫生研究院 (NIH) 资助。