Assembly of the Bacteriophage T4 Primosome

噬菌体 T4 引发体的组装

• 批准号: 6879044
• 负责人: Michelle Marie Spiering
• 金额: $ 4.83万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6879044
• 关键词: DNA binding protein; DNA footprinting; DNA replication; bacteriophage T4; cell line; electron microscopy; fluorescence resonance energy transfer; postdoctoral investigator; protein protein interaction; protein structure function; protein transport; virus assembly; virus genetics; virus protein

DESCRIPTION (provided by applicant): The bacteriophage T4 replication system is one of the best-studied of the related mechanisms of DNA replication employed by prokaryotes and eukaryotes. This proposal is focused on the assembly and functioning of the primosome in the context of DNA replication. A complete understanding will require studying the individual proteins of the primosome as a large enzyme machine. The assembly process of the primosome will be interrogated by ensemble and single-molecule FRET equilibrium and pre-steady state studies. Additional structural information on the primosome will be gained by electron microscopy. The functioning of the primosome will by elucidated by cross-linking/footprinting studies and by monitoring protein-protein interactions using FRET during active translocation and priming by the primosome. Completion of this project will result in a description of the assembly and functioning of the primosome in detail similar to that already determined for the polymerase holoenzyme. Since the process of DNA replication has profound implications for human disease, including cancer, understanding the fidelity of DNA replication is of significant humanitarian and economic interest.

描述（由申请人提供）： 噬菌体T4复制系统是原核生物和真核生物采用的DNA复制机制中最好的研究之一。该建议的重点是在DNA复制背景下原始体的组装和功能。完全理解将需要研究原始体的单个蛋白质作为大型酶机。原始体的组装过程将通过集合和单分子fret平衡和稳态的状态研究来审问。关于原始体的其他结构信息将通过电子显微镜获得。通过交联/足迹研究阐明原始体的功能，并通过在活性易位和原始体启动过程中使用FRET监测蛋白质 - 蛋白质相互作用。该项目的完成将导致对原始体的组装和功能的描述，类似于已经确定的聚合酶全酶。由于DNA复制过程对包括癌症在内的人类疾病具有深远的影响，因此了解DNA复制的忠诚度具有重要的人道主义和经济利益。