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（由申请人提供）：遗传不稳定性与突变、染色体重排和非整倍体的发生率增加有关。这是癌症的标志，也是其他重大健康挑战的标志，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复制应激是基因组不稳定性的关键因素。稳定复制叉、防止异常分裂和促进DNA修复的分子机制是疾病的主要屏障;因此，了解它们的功能与人类健康直接相关。该建议采用已建立的模型遗传系统来鉴定和表征维持基因组稳定性的分子途径。该项目使用遗传学、分子生物学和新的细胞生物学方法在裂殖酵母S。粟酒裂殖酵母，以检查细胞在正常营养细胞周期和减数分裂期间对复制应激的反应。S.粟酒裂殖酵母是一种成熟的染色体生物学模型，与人类细胞具有许多共同特征。该提案研究了这样一种假设，即对复制应激的反应动力学决定了细胞是否阻止细胞周期，或者它们是否逃避正常的检查点并继续异常分裂，产生染色体重排和突变率增加。复制压力可能会在整个基因组和研究的一个重要组成部分是作为一个模型脆弱的网站pericentromere的分析。另一个新的组成部分是分析减数分裂过程中的复制压力，作为与出生缺陷和不育相关的染色体重排的贡献者。一个重要的创新是活细胞谱系的新系统 分析与定量分析相结合，以研究对损坏和检查点规避的动态响应。这种活的全细胞分析允许识别经历不同结果的不同细胞亚群，从而产生与多种疾病相关的不稳定循环。通过将这种新的细胞生物学方法与高超的酵母基因发现工具相结合，并识别导致异常分裂和进一步应激的分子事件，该项目将解决当前理解中的关键空白。哪些途径导致对压力的不同反应及其相关病理？它们是如何彼此不同以产生不同的结果的，例如簇状突变、CNV、缺失和重复以及染色体重排？总之，这些研究提供了保守蛋白如何相互作用以维持真核细胞中基因组稳定性的整体图景，确定了人类疾病的标志物和风险因素。