Maintaining the integrity of a genome

维持基因组的完整性

• 批准号: 10672392
• 负责人: JENNIFER L GERTON
• 金额: $ 36.99万
• 依托单位: STOWERS INSTITUTE FOR MEDICAL RESEARCH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10672392
• 关键词: 3-Dimensional; Address; Affect; Amaze; Anaphase; Aneuploidy; Binding; Biology; Calibration; Cancer Etiology; Catenated DNA; Cell Culture Techniques; Cells; Centromere; ChIP-seq; Chromosomal Instability; Chromosome 4; Chromosome Segregation; Chromosome Structures; Chromosomes; Complement; Complex; Cultured Cells; DNA; Data; Double Strand Break Repair; Elements; Ensure; Environment; Event; Fatigue; Genetic; Genetic Determinism; Genome; Genome Stability; Genomic Instability; Goals; Hand; Haplotypes; Health; Human; Human Cell Line; Human Chromosomes; Human Genome; Human Pathology; Image; Imaging Device; Individual; Kinetochores; Literature; Malignant Neoplasms; Maps; Mediating; Metaphase; Microtubules; Mission; Mitosis; Modeling; Molecular; Mutation; Organizational Models; Outcome; Pathologic; Pattern; Persons; Proteins; Public Health; Repetitive Sequence; Reporting; Research; Role; Shapes; Sister Chromatid; Testing; Time; Topoisomerase II; Tumor Tissue; Variant; Work; Yin-Yang; blind; cancer type; chromosome loss; chromosome missegregation; cohesin; cohesion; genetic information; genome integrity; genomic tools; grasp; human tissue; innovation; loss of function; micronucleus; molecular modeling; prevent; reproductive senescence; segregation; superresolution imaging; tissue/cell culture; tool; transmission process; tumor xenograft

Project Summary/Abstract In order to fully grasp the molecular origins of genome instability, the field must understand at a molecular level how centromeres work to promote the stable transmission of chromosomes. Genome instability underlies a variety of human pathologies, including cancer and reproductive aging. Our long-term goal is to determine how the evolutionarily conserved cohesin complex maintains genome integrity through its roles in chromosome segregation, chromosome organization, and double-strand break repair. Loss of sister chromatid cohesion is speculated to be a major contributor to chromosome instability. The objective of this application is to produce a molecular model for how cohesin operates at individual human centromeres to achieve centromeric cohesion and accurate chromosome segregation. The central hypothesis is that cohesin and DNA catenation together create centromere-unique landscapes of sister chromatid cohesion to prevent chromosome instability. The variation in human centromeres and centromeric cohesion may therefore impact the transmission of each chromosome. We will test the idea that centromere-specific cohesion must be considered as a genetic determinant of sister chromatid cohesion and segregation in order to have a complete model for how chromosomal instability occurs through two specific aims: 1) discover the landscape of centromeric cohesion at individual human centromeres and 2) examine how chromosome centromeric cohesion maintains euploidy. Under the first aim, calibrated paired-end ChIP seq will be used to map cohesin binding relative to kinetochore proteins and human centromeric arrays in human tissue culture cells. This approach will be complemented by superresolution imaging of the same three components (centromeres, kinetochores, and cohesin) in cells, and will include imaging-based determination of centromere-specific cohesion. Together these approaches will produce a linear and 3D map of cohesion within and around individual human centromeres. In the second aim we will examine how centromere-specific patterns of centromeric cohesion prevent chromosome missegregation events in cultured cells and in xenograft tumor tissue. The outcome will be fundamental principles of centromeric array-based cohesion fatigue and resulting patterns of chromosome instability. The research is innovative because it incorporates the latest information on human centromeric DNA arrays, a new working model for the organization of centromeric DNA by cohesion, and new quantitative molecular, genomic, and imaging tools to probe how centromeric cohesion enforces accurate sister chromatid segregation. The proposed research is significant because centromeric arrays may be unrecognized genetic determinants of chromosome instability. Many types of cancer are associated with seemingly random patterns of instability that may have molecular origins in unique centromeric cohesion profiles. Furthermore, many cancers are associated with mutations that impact chromosome segregation machinery, such as cohesin. The outcome of this project will be a more complete picture of the mechanisms underlying chromosomal instability.

项目摘要/摘要 为了完全掌握基因组不稳定性的分子起源，该场必须在分子水平上理解 中心粒如何促进染色体的稳定传播。基因组不稳定性是 各种人类病理，包括癌症和生殖衰老。我们的长期目标是确定如何 进化保守的粘着素复合物通过其在染色体中的作用保持基因组完整性 隔离，染色体组织和双链断裂修复。姐妹染色单体内聚力的损失是 推测是染色体不稳定性的主要因素。该应用的目的是生产 分子模型的粘着蛋白如何在单个人的centromeres上运行以实现丝粒内聚会 和准确的染色体分离。中心假设是粘蛋白和DNA融合在一起 创建姐妹染色单体内聚力的Centromere唯一景观，以防止染色体不稳定性。这 因此，人类的丝粒和丝粒凝聚力的变化可能会影响每个的传播 染色体。我们将测试以下思想，即必须将Centromere特异性内聚力视为遗传 姐妹染色单体内聚和隔离的决定因素，以便有一个完整的模型 染色体不稳定性通过两个特定目的发生：1）在 单个人体centromeres和2）检查染色体centromeric凝聚力如何保持植物。 在第一个目标下，校准的配对芯片SEQ将用于映射粘蛋白的结合相对于动力学 人类组织培养细胞中的蛋白质和人类丝粒阵列。这种方法将得到补充 相同三个成分（centromeres，sbinetochores和obhesin）的上分辨率成像，并且 将包括基于成像的共粒特异性内聚力的确定。这些方法将在一起 在单个人类的centromeres内和周围产生线性和3D图。在第二个目标 我们将研究丝粒特异性的丝粒凝聚力模式如何预防染色体 培养细胞和异种移植肿瘤组织中的错误分析事件。结果将是基本的 基于丝粒阵列的原理基于染色体不稳定性的凝聚力和产生的模式。这 研究之所以创新，是因为它包含了有关人类丝粒DNA阵列的最新信息，这是一个新的 通过内聚力和新的定量分子基因组，用于组织丝粒DNA的工作模型 和成像工具，以探测中心粒的内聚力如何实现准确的姐妹染色质被隔离。这 拟议的研究很重要，因为中心层阵列可能是未被认可的遗传决定因素 染色体不稳定性。许多类型的癌症与看似随机的不稳定模式有关， 可能具有分子起源于独特的丝粒内聚谱。此外，许多癌症是 与影响染色体分离机械的突变有关，例如粘着蛋白。结果 该项目将是染色体不稳定性基础机制的更完整图片。