Structural Studies of the T7 DNA Replisome

T7 DNA 复制体的结构研究

• 批准号: 6487900
• 负责人: YING LI
• 金额: $ 3.83万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-25 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6487900
• 关键词: DNA directed DNA polymerase; DNA primase; DNA replication; bacteriophage T7; chemical structure function; computer data analysis; cryoelectron microscopy; enzyme substrate complex; helicase; image enhancement; intermolecular interaction; model design /development; molecular assembly /self assembly; molecular site; nucleic acid structure; physical model; postdoctoral investigator; protein purification; structural biology; virus DNA; virus protein; virus replication

DESCRIPTION: (provided by applicant) The stability of the genome, which is dependent on faithful DNA replication, is essential to normal cell functions. Most human cancers have been shown to be related to some forms of genetic instabilities. Therefore, understanding the mechanism of DNA replication is fundamental in understanding the underlying causes of DNA instability related human diseases, as well as developing potential diagnostic and therapeutic approaches for these diseases. The objective of this research is to study the global organization of a replication fork, as the first step in understanding the structural basis for the coordinated synthesis of the leading and lagging strands during DNA replication. The bacteriophage T7 DNA replication system will be used as the model system in this study. Electron cryo-microscopic approaches will be employed to determine the structures of the complexes formed between replisomal proteins. The specific aims of this proposal are: 1. Determine the structures of the T7 gene 4 helicase/ primase and the T7 DNA polymerase complexes in the contexts of the leading and lagging strand synthesis, respectively, using electron cryomicroscopic (cryoEM) methods. 2. Determine the structure of the entire T7 phage replisome using cryoEM methods.

描述：（由申请人提供）基因组的稳定性，即