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.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 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）作为了解转录机制的模型系统。我们通过使用Semet蛋白MAD方法和在X25/NSLS获得的数据集以2.3A分辨率解决了APO酶结构。最近，我们通过分子替代求解了N4 RNAP-DNA复合物结构。该数据集用Home Source X射线收集，分辨率为2.4a。没有人在转录启动阶段（转录起始复合物）解决RNAP-DNA-SUBSTRATE复合物的结构。我们通过共结晶制备了启动复合物晶体，并发现它以2.4A分辨率的衍射与Home Source X射线分辨率。为了了解转录模仿反应的分子细节，我们希望要求F1/国际象棋的光束线时间以原子分辨率收集转录起始复杂的晶体数据集。该结构将是转录启动阶段的第一个快照，它告诉我们RNAP为什么能够进行从头反应。该项目由NIH资助。