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: 10595031
• 负责人: SUSAN L FORSBURG
• 金额: $ 70.73万
• 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 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10595031
• 关键词: 3-Dimensional; Address; Affect; Aging; Biological; Biology; Cell model; Cell physiology; Cells; Cellular biology; Centromere; Characteristics; Chromosomal Stability; Chromosome Fragile Sites; Chromosome Segregation; Chromosome Structures; Chromosomes; Congenital Abnormality; Cytoprotection; DNA; DNA Repair; DNA Sequence; DNA biosynthesis; DNA replication fork; Data; Defect; Development; Disease; Ensure; Eukaryotic Cell; Event; Fission Yeast; Genes; Genetic; Genetic Models; Genome Stability; Genomic Instability; Goals; Health; Human; Infertility; Link; Malignant Neoplasms; Meiosis; Mitosis; Molecular; Neurologic; Normal Cell; Organism; Pathology; Pathway interactions; Phase; Proteins; Resolution; Ribosomal DNA; Risk Factors; Role; Stress; Structure; Systems Biology; Yeasts; biomarker identification; developmental disease; gene discovery; genetic information; human disease; live cell imaging; mutant; nervous system disorder; novel; preservation; prevent; repaired; replication stress; response; tool; transmission process

Genome instability refers to changes in chromosome sequence, structure, or number that affect normal cell function. Such instability is characteristic of cancer, as well as certain developmental and neurological defects, and aging. Data from multiple organisms suggests that DNA replication stress is a key contributor to genome instability. Mechanisms that stabilize replication forks, prevent abnormal divisions, and promote DNA repair are a primary barrier to disease; therefore, understanding their function and the consequences of their disruption has direct relevance to human health. This proposal employs an established model cell biology system, the fission yeast S. pombe, to characterize how living cells respond to replication stress. S. pombe is a well-established genetic model for chromosome biology that shares many features with human cells. Significantly, nearly all the genes under study have orthologues in humans that have been associated with disease.. A key aspect of the approach is to use live cell imaging to characterize the response to replication stress and characterize its long term consequences. The overarching goal is to understand the dynamics of replication stress and its resolution in normal and mutant cells. This includes determining how the cell deploys molecular mechanisms to allow damage resolution and ensure chromosome segregations. We address the cellular and genetic consequences of division under stress; investigate how replication occurs late in G2 or mitosis to facilitate resolution; and examine the three-dimensional organization of repair structures. We have previously shown that the pericentromere is a fragile site, and we have expanded that to examine the ribosomal DNA and the role of phase separation in contributing to gene integrity, as well as identification of other fragile regions. A novel component is the analysis of replication stress during meiosis as a contributor to chromosome rearrangements associated with birth defects and infertility. By combining this cell biological approach with superb yeast gene-discovery tools, and identifying the molecular events that lead to abnormal divisions and further stress, this project tackles a critical gap in current understanding. What are the pathways that contribute to different responses to stress and their associated pathologies and how do they affect the biology of living cells? 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.

基因组不稳定性是指影响染色体序列、结构或数量的变化。 正常的细胞功能。这种不稳定性是癌症的特征，也是某些发育和 神经系统缺陷和衰老。来自多个生物体的数据表明 DNA 复制压力是 基因组不稳定的关键因素。稳定复制叉、防止异常的机制 分裂和促进 DNA 修复是疾病的主要屏障；因此，了解他们的 其功能及其破坏的后果与人类健康直接相关。 该提案采用了已建立的模型细胞生物学系统，即裂殖酵母 S. pombe， 表征活细胞如何响应复制压力。粟酒裂殖酵母是一种成熟的遗传 与人类细胞具有许多共同特征的染色体生物学模型。值得注意的是，几乎所有 正在研究的基因在人类中具有与疾病相关的直系同源物。 该方法的一个关键方面是使用活细胞成像来表征对 复制压力并描述其长期后果。总体目标是了解 正常和突变细胞中复制应激的动态及其解决。这包括 确定细胞如何部署分子机制来解决损伤并确保 染色体分离。我们解决分裂的细胞和遗传后果 压力;研究复制如何在 G2 晚期或有丝分裂中发生以促进解决；并检查 修复结构的三维组织。我们之前已经表明，着丝粒周围 是一个脆弱位点，我们已将其扩展以检查核糖体 DNA 和相的作用 分离有助于基因完整性，以及识别其他脆弱区域。一本小说 成分是减数分裂过程中复制应激作为染色体贡献者的分析 与出生缺陷和不孕症相关的重排。 通过将这种细胞生物学方法与卓越的酵母基因发现工具相结合， 该项目确定了导致异常分裂和进一步压力的分子事件，解决了 当前理解中的重大差距。导致不同反应的途径是什么 压力及其相关病理以及它们如何影响活细胞的生物学？一起， 这些研究提供了保守蛋白质如何相互作用以维持基因组稳定性的整体图景 在真核细胞中，识别人类疾病的标记和危险因素。