Defining the effect of centromere/kinetochore associations on genome instability

项目摘要 体细胞拷贝数改变（SCNA）是癌症的一个既定标志，并且是由基因组中的错误引起的。 维持基因组稳定的机制。SCNA在大多数癌症类型中的存在突出了 预防SCNA作为预防癌症产生和传播的潜在策略。预防 SCNAs需要维持基因组的稳定性，这取决于基因组的正常功能。 着丝粒着丝粒是重复的DNA结构域，是动粒组装的位点， 将着丝粒与微管结合并调节染色体分离的多蛋白复合物。 有趣的是，着丝粒富含重复序列的结构使它们易于发生基因组重排， 癌细胞中40-60%的基因组重排发生在着丝粒。因此，着丝粒维持着 基因组稳定性和自身的遗传完整性。癌症起始和 进展总是涉及某种形式的基因组不稳定性，导致SCNA，因此机制 着丝粒DNA/动粒蛋白质相互作用对基因组稳定性的作用是一种 研究不足但却是癌症研究的关键组成部分。这个项目的目标是确定如何改变 在着丝粒-动粒界面影响基因组稳定性，长期目标是发现新的 基因组不稳定性导致癌症发生和发展的机制。这是一个新的建议，因为 这是第一个测试着丝粒DNA和相关蛋白质多样性对基因组的机制作用的实验。 稳定我假设，着丝粒的变化可以影响着丝粒/动粒联合， 随后影响基因组稳定性。目的1将确定是否CENP-A，一个着丝粒特异性组蛋白， H3变异体，与种内不同的着丝粒卫星重复序列差异相关，表明 着丝粒DNA重复序列与CENP-A结合以维持其稳定性的差异能力。目标2将 确定着丝粒序列相容性对动粒蛋白功能的依赖性。这将是 通过观察人类Shugoshin 1的功能来完成，Shugoshin 1是一种与着丝粒相关的动粒蛋白质 在有丝分裂过程中维持着丝粒粘连蛋白的DNA，带有小鼠着丝粒卫星重复序列。所有这些 aims将强调依赖于着丝粒序列发挥功能的途径， 着丝粒依赖机制来维持基因组稳定性。我的训练目标是（i）获得 实验技能，使我能够使用细胞培养系统来测试计算生成的 （2）提高科学沟通能力。这些将有助于我的事业 发展成为一名独立的学术科学家，研究进化遗传学的作用。 人类癌症发病率和演变的过程。我的研究生课程和机构， 培训记录，两个优秀的赞助商将为我提供强大的培训和资源，以执行 研究培训计划，最终帮助我实现目标。

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.