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复制和维护对于人类健康至关重要。 由于内源性或环境攻击引起的DNA损害可能会阻碍DNA的复制，并且 诱导基因组异常和人类疾病。重新构体是负责的多蛋白质复合物 用于DNA复制。此外，作为第一个遇到任何复制障碍的人，重新播放戏剧 在信号复制应力和促进DNA修复中的基本作用。但是，结构基础 重新组合操作和重新介导的复制应力反应和修复尚不清楚。 我们建议使用生化重构和冷冻EM结构决心来研究如何 重置体在正常和压力条件下起作用 遗传变异和人类疾病。首先，我们将利用噬菌体T7的重建体作为 模型系统以研究在压力条件下重新构体操作的结构基础。我 最近获得了在其DNA底物上运行的T7重建体的第一个结构，该结构 说明了组织的组织和工作机制。我们将进一步调查如何 T7对各种复制壁垒的反应，以触发适当的复制耦合 维修。其次，我们将研究线粒体DNA复制的结构基础。线粒体 是电池的动力室，并且由于DNA复制故障而导致的线粒体DNA上的错误是 与许多人类遗传疾病有关。我们将重建线粒体重置和 研究线粒体DNA的复制方式，以及线粒体上的突变如何导致 人类疾病。我们的调查将为理解提供第一个结构框架 重新介导的DNA复制，应力反应和修复。结构信息将 有助于开发新的疗法，以靶向或避免靶向DNA复制。