Structural basis of replisome mediated DNA replication and repair

复制体介导的 DNA 复制和修复的结构基础

• 批准号: 10275032
• 负责人: Yang Gao
• 金额: $ 38.43万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10275032
• 关键词: Bacteriophage T7; Biochemical; Biological Models; Cells; Coupled; Cryoelectron Microscopy; DNA; DNA Maintenance; DNA Repair; DNA Sequence Alteration; DNA biosynthesis; Development; Genetic Diseases; Genetic Variation; Genome; Genomic DNA; Health; Human; Human Genetics; Investigation; Lead; Life; Malignant Neoplasms; Mediating; Mitochondria; Mitochondrial DNA; Multiprotein Complexes; Mutation; Play; Proteins; Role; Signal Transduction; Stress; Structure; assault; biological adaptation to stress; human disease; novel therapeutics; operation; reconstitution; repaired; replication stress; response

DNA is the blueprint of life and DNA replication and maintenance are essential for human health. Damages on DNA due to endogenous or environmental assaults can hamper DNA replication and induce genome abnormities and human diseases. A replisome is a multi-protein complex responsible for DNA replication. Moreover, as the first one to meet any replication barrier, the replisome plays essential roles in signaling replication stress and facilitating DNA repair. However, the structural basis of replisome operation and replisome mediated replication stress response and repair remains unclear. We propose to use biochemical reconstitution and cryo-EM structural determination to investigate how the replisome acts under normal and stressed conditions and how disfunction of the replisome leads to genetic variations and human diseases. First, we will employ replisome from bacteriophage T7 as a model system to investigate the structural basis of replisome operation under stressed conditions. I have recently obtained the first structure of the T7 replisome operating on its DNA substrate, which illustrated the organization and the working mechanism of a replisome. We will further investigate how the T7 replisome responses to various replication barriers to trigger appropriate replication-coupled repair. Second, we will investigate the structural basis of mitochondria DNA replication. Mitochondria is the power house of a cell and errors on mitochondria DNA due to faulty DNA replication are connected to many human genetic diseases. We will reconstitute the mitochondria replisome and investigate how mitochondria DNA is replicated and how mutations on mitochondria replisome lead to human diseases. Our investigations will provide the first structural framework for understanding replisome mediated DNA replication, stress response and repair. The structural information will contribute to the development of novel therapies to target or avoid targeting DNA replication.

DNA是生命的蓝图，DNA的复制和维护对人类健康至关重要。