Mechanisms linking replication stress to genome instability in fission yeast

裂殖酵母中复制应激与基因组不稳定性的联系机制

• 批准号: 9893001
• 负责人: SUSAN L FORSBURG
• 金额: $ 68.67万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2021-05-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9893001
• 关键词: Aging; Aneuploidy; Biological; Biology; Cell Cycle; Cell Cycle Arrest; Cells; Cellular biology; Chromosomal Stability; Chromosome Fragile Sites; Chromosomes; Congenital Abnormality; Coupled; DNA; DNA Repair; DNA biosynthesis; DNA replication fork; Data; Defect; Development; Disease; Ensure; Eukaryotic Cell; Event; Fission Yeast; Genes; Genetic; Genetic Materials; Genetic Models; Genome; Genome Stability; Genomic Instability; Health; Human; Infertility; Lead; Link; Malignant Neoplasms; Meiosis; Methods; Modeling; Molecular; Molecular Biology; Mutation; Organism; Outcome; Pathology; Pathway interactions; Proteins; Risk Factors; Stress; System; Yeasts; gene discovery; genetic information; genetic pedigree; human disease; innovation; nervous system disorder; novel; preservation; prevent; public health relevance; replication stress; response; tool; transmission process

 DESCRIPTION (provided by applicant): Genetic instability is associated with increased rates of mutation, chromosome rearrangement, and aneuploidy. This is a hallmark of cancer, as well as other significant health challenges including developmental defects, neurological disorders, and aging. Data from multiple systems suggests that DNA replication stress is a key contributor to genome instability. Molecular mechanisms that stabilize replication forks, prevent abnormal divisions, and promote DNA repair are a primary barrier to disease; therefore, understanding their function has direct relevance to human health. This proposal employs an established model genetic system to identify and characterize molecular pathways that maintain genome stability. The project uses genetics, molecular biology, and novel cell biology methods in the fission yeast S. pombe to examine the response of cells to replication stress during the normal vegetative cell cycle, and during meiosis. S. pombe is a well-established model for chromosome biology that shares many features with human cells. The proposal investigates the hypothesis that the dynamics of the response to replication stress determines whether cells arrest the cell cycle, or whether they evade normal checkpoints and go on to divide abnormally, generating chromosome rearrangements and increased rates of mutation. Replication stress may vary across the genome and a significant component of the study is the analysis of the pericentromere as a model fragile site. An additional novel component is the analysis of replication stress during meiosis as a contributor to chromosome rearrangements associated with birth defects and infertility. A significant innovation is a new system for live cell pedigree analysis coupled with quantitative analysis to investigate the dynamic response to damage and checkpoint evasion. This live whole-cell analysis allows the identification of distinct sub-populations of cells that undergo different outcomes creating a cycle of instability that is associated with multiple diseases By combining this new cell biological approach with superb yeast gene-discovery tools, and identifying the molecular events that lead to abnormal divisions and further stress, this project will tackle a critical gap in current understanding. What are the pathways that contribute to different responses to stress and their associated pathologies? How do they differ from one another to generate distinct outcomes such as clustered mutations, CNVs, deletions and duplications, and chromosome rearrangements? Together, these studies provide a holistic picture of how conserved proteins interact to maintain genome stability in a eukaryotic cell, identifying markers and risk factors for human disease.

 描述（由适用提供）：遗传不稳定性与突变率提高，染色体重排和非整倍性有关。这是癌症的标志，也是其他重大健康挑战，包括发育缺陷，神经系统疾病和衰老。来自多个系统的数据表明，DNA复制应力是导致基因组不稳定性的关键因素。稳定复制叉，预防异常分裂和促进DNA修复的分子机制是疾病的主要障碍。因此，了解其功能与人类健康有直接相关。该建议采用已建立的模型遗传系统来识别和表征维持基因组稳定性的分子途径。该项目在裂变酵母菌中使用遗传学，分子生物学和新型细胞生物学方法，以检查细胞对正常植物细胞S. pombe在正常植物性细胞中复制应激的反应。该提案调查了以下假设：对复制应力的反应动力学决定了细胞是否阻止了细胞周期，还是逃避正常检查点并继续对异常分裂，从而产生染色体重排和更大的突变速率。在整个基因组中，复制应力可能会有所不同，研究的重要组成部分是将围粒子作为模型脆弱部位的分析。另一个新的组成部分是对减数分裂过程中复制应力的分析，是导致与先天缺陷和不育相关的染色体重排的贡献者。重大创新是实时细胞谱系的新系统 分析结合定量分析，以研究对损伤和检查点演变的动态响应。这种实时的全细胞分析允许鉴定细胞的不同亚种群，这些细胞经历了不同的结果，从而产生了不稳定性的循环，该循环通过将这种新的细胞生物学方法与超级酵母基因分离工具相结合，与多种疾病相关，并识别出异常的划分和进一步的压力，该项目将在当前的理解中差距。有助于对压力及其相关病理的不同反应的途径是什么？它们如何彼此不同，以产生不同的结果，例如聚类突变，CNV，缺失和重复以及染色体重排？这些研究共同提供了整体图片，说明成本蛋白如何相互作用以维持真核细胞中的基因组稳定性，从而确定了人类疾病的标记和危险因素。