Structure-Function Analysis of DNA Replication Initiation Factors Implicated in Disease

与疾病相关的 DNA 复制起始因子的结构功能分析

• 批准号: 10707959
• 负责人: Gheorghe Chistol
• 金额: $ 38.62万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-21 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10707959
• 关键词: Biochemical; Biological Markers; Cell Proliferation; DNA analysis; DNA biosynthesis; DNA-Protein Interaction; Disease; Genetic; Genetic Diseases; Genome; Genome Stability; Human; Laboratories; Life; Malignant Neoplasms; Methods; Molecular; Mutation; Oncogene Activation; Organism; Peptide Initiation Factors; Process; Proliferating; Proteins; Regulation; Replication Initiation; Replication Origin; Role; Structure; Time; Visualization; Work; Yeasts; cancer cell; genome integrity; imaging platform; insight; new therapeutic target; novel; overexpression; rare genetic disorder; single molecule

PROJECT SUMMARY DNA replication is a highly conserved process essential for all life. Since eukaryotic genomes are very large, they are replicated in parallel from thousands of replication origins. Proper regulation of replication initiation is essential for maintaining genome stability. Oncogene activation dysregulates replication initiation and enables rapid cell proliferation in cancer. Key regulators of replication initiation are over-expressed in cancer, and mutations in these proteins cause rare genetic disorders. Yet, the molecular mechanisms that underlie replication initiation remain poorly understood, especially in multicellular organisms, for two main reasons: (1) replication initiation involves transient protein-protein and protein-DNA interactions that are very challenging to study using existing biochemical, genetic, and structural methods; and (2) the most comprehensive studies of replication initiation have been done in yeast, however key regulators of replication initiation are poorly conserved between yeast and metazoans. Notably, some human replication factors are 2-3x larger than their yeast counterparts and containing novel domains with unknown functions. My laboratory recently developed a single-molecule imaging platform to visualize replication initiation in real time. We will use this powerful approach to (i) provide a quantitative description of replication initiation; (ii) understand how this process changed from yeast to humans; (iii) delineate the role of key replication proteins and their domains that are unique to metazoans; and (iv) understand why mutations in some replication initiation factors cause disease. Our work will also provide valuable mechanistic insights into how cancer cells sustain rapid proliferation by over-activating replication initiation and inform how replication initiation factors may be used as biomarkers or targets for novel therapies.

项目总结