Defining the effect of centromere/kinetochore associations on genome instability

定义着丝粒/着丝粒关联对基因组不稳定性的影响

• 批准号: 10385959
• 负责人: Uma Arora
• 金额: $ 4.25万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-16 至 2022-12-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10385959
• 关键词: Address; Affect; Affinity; Amino Acids; Aneuploidy; Architecture; Binding; Biological; Cell Culture System; Cell Culture Techniques; Cells; Centromere; ChIP-seq; Chromatin; Chromosome Segregation; Communication; Complex; Computer Analysis; Core Facility; Couples; DNA; DNA Sequence Rearrangement; Data; Databases; Dependence; Ensure; Epigenetic Process; Evolution; Exhibits; Experimental Genetics; Foundations; Generations; Genetic; Genetic Processes; Genetic Variation; Genome; Genome Stability; Genomic Instability; Genomics; Goals; Histone H3; House mice; Human; Inbred Strains Mice; Inbreeding; Incidence; Institution; Kinetochores; Knowledge; Lead; Learning; Left; Light; Maintenance; Malignant Neoplasms; Mammals; Measures; Methods; Microtubules; Mitosis; Mitotic; Molecular; Mouse Strains; Multiprotein Complexes; Mus; Orthologous Gene; Pathway interactions; Prevention; Proteins; Regulation; Research; Research Training; Resources; Role; Scientist; Series; Signal Transduction; Site; Testing; Time; Training; Training Programs; Transgenic Organisms; Variant; Work; anticancer research; base; cancer cell; cancer initiation; cancer type; career development; centromere protein A; cohesin; cohort; embryonic stem cell; epigenomics; experimental study; functional genomics; improved; novel; prevent; programs; protein function; skills; therapeutic target; tool; transgene expression; tumor progression; tumorigenesis

PROJECT SUMMARY Somatic copy number alterations (SCNAs) are an established hallmark of cancer and result from errors in the mechanisms that maintain genome stability. The presence of SCNAs across most cancer types highlights prevention of SCNAs as a potential strategy to prevent the generation and propagation of cancer. Prevention of SCNAs requires the maintenance of genome stability, which is dependent on the normal functioning of centromeres. Centromeres, which are repetitive DNA domains, are the site of assembly of the kinetochore, a multi-protein complex that couples centromeres with microtubules and regulates chromosome segregation. Interestingly, the repeat-rich architecture of centromeres makes them prone to genomic rearrangements, with 40-60% of genomic rearrangements in cancer cells occurring at centromeres. Hence, centromeres maintain genome stability and their own genetic integrity by associating with kinetochore proteins. Cancer initiation and progression always involve some form of genome instability that leads to SCNA, hence the mechanisms through which centromere DNA/kinetochore protein interactions contribute to genome stability are an understudied but critical component of cancer research. The goal of this project is to determine how changes at the centromere-kinetochore interface impact genome stability, with the long-term goal being to uncover new mechanisms of genome instability leading to cancer initiation and progression. This proposal is novel because it is the first to test the mechanistic effect of centromere DNA and associated protein diversity on genome stability. I hypothesize that centromere variation can influence centromere/kinetochore association and subsequently impact genome stability. Aim 1 will determine whether CENP-A, a centromere-specific histone H3 variant, differentially associates with divergent centromere satellite repeats within species, indicating differential ability of centromere DNA repeats to associate with CENP-A to maintain their stability. Aim 2 will determine the dependence of kinetochore protein function on centromere sequence compatibility. This will be done by observing the function of human Shugoshin 1, a kinetochore protein that associates with centromere DNA to maintain centromeric cohesin during mitosis, with mouse centromere satellite repeats. Together, these aims will highlight pathways that are dependent on centromere sequence for their function, implicating centromere-dependent mechanisms to maintain genome stability. My goals for training are (i) to gain an experimental skill set that will enable me to use a cell culture system to test computationally generated hypotheses and (ii) to improve my scientific communication skills. These will contribute to my career development to become an independent academic scientist investigating the role of evolutionary genetic processes in human cancer incidence and evolution. My graduate program and institution with a successful training record, and two excellent sponsors will provide me with strong training and resources to execute the research training plan and ultimately help me achieve my goals.

项目摘要 体拷贝数改变（SCNA）是癌症的既定标志，是由于错误的错误而导致的 维持基因组稳定性的机制。大多数癌症类型的SCNA的存在突出显示 预防SCNA是防止癌症产生和传播的潜在策略。预防 SCNA需要维持基因组稳定性，这取决于正常功能 中心。是重复的DNA域的丝粒，是动元组装的位置，A 多蛋白质复合物将与微管融合并调节染色体分离。 有趣的是，centromeres的重复构建使它们容易重新排列，并带有 癌细胞中的40-60％的基因组重排发生。因此，中心粒保持 基因组稳定性及其自身的遗传完整性通过与动力学蛋白相关联。癌症的启动和 进展始终涉及某种形式的基因组不稳定性，导致SCNA，因此机理 Centromere DNA/动力学蛋白相互作用有助于基因组稳定性是一种 癌症研究的研究研究但至关重要。该项目的目标是确定如何改变 在Centromere-Kinetochore界面影响基因组稳定性，长期目标是发现新的 基因组不稳定性的机制导致癌症的开始和进展。该建议是新颖的，因为 它是第一个测试中心粒DNA和相关蛋白多样性对基因组的机械作用的一种 稳定。我假设中心粒的变化会影响中心粒/动物学的关联和 随后影响基因组稳定性。 AIM 1将确定CENP-A是否是丝粒特异性组蛋白 H3变体，将差异化与物种内部的丝粒卫星重复相关联，表明 CentRomere DNA重复与CENP-A保持稳定性的差异能力。 AIM 2意志 确定动力学蛋白功能对中心粒序列兼容性的依赖性。这将是 通过观察人类Shugoshin 1的功能，该功能是一种与Centromere相关的动力学蛋白 DNA在有丝分裂过程中维持着丝粒粘着蛋白，并具有小鼠丝粒卫星重复序列。在一起，这些 目的将突出显示依赖着丝粒序列的途径，这意味着 依赖着基因组稳定性的Centromere依赖机制。我的培训目标是（i） 实验技能集将使我能够使用细胞培养系统测试计算生成 假设和（ii）提高我的科学沟通技巧。这些将有助于我的职业 发展成为一名独立学术科学家，研究进化遗传的作用 人类癌症发病率和进化过程中的过程。我的研究生课程和机构成功 培训记录和两个出色的赞助商将为我提供强大的培训和资源来执行 研究培训计划，最终帮助我实现自己的目标。