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影响的染色体序列，结构或数字的变化 正常的细胞功能。这种不稳定性是癌症的特征，以及某些发育和 神经系统缺陷和衰老。来自多种生物的数据表明DNA复制应力是 基因组不稳定性的关键因素。稳定复制叉的机制，预防异常 分裂和促进DNA修复是疾病的主要障碍。因此，了解他们的 功能及其破坏的后果与人类健康直接相关。 该建议采用已建立的模型细胞生物学系统，裂变酵母菌S. Pombe， 表征活细胞如何应对复制应力。 S. Pombe是一个完善的遗传 染色体生物学模型，与人类细胞具有许多特征。重要的是，几乎所有 研究的基因在与疾病有关的人类中有直系同源物。 该方法的一个关键方面是使用活细胞成像来表征对 复制应力并表征其长期后果。总体目标是了解 复制应力的动力学及其在正常细胞和突变细胞中的分辨率。这包括 确定细胞如何部署分子机制以允许损害解决并确保 染色体隔离。我们解决了分裂下的细胞和遗传后果 压力;研究如何在G2或有丝分裂中进行复制以促进分辨率；并检查 维修结构的三维组织。我们以前已经表明围粒粒粒 是一个脆弱的部位，我们已经将其扩展为检查核糖体DNA和相位的作用 促进基因完整性以及对其他脆弱区域的识别的分离。小说 组成部分是减数分裂过程中复制应力的分析作为染色体的促成 与先天缺陷和不育有关的重排。 通过将这种细胞生物学方法与出色的酵母基因发现工具相结合，并 确定导致异常分裂和进一步压力的分子事件，该项目解决了 当前理解中的关键差距。有助于不同的回答的途径是什么 压力及其相关的病理以及它们如何影响活细胞的生物学？一起， 这些研究提供了整体图片，说明保守蛋白如何相互作用以维持基因组稳定性 在真核细胞中，确定人类疾病的标记和危险因素。