Functional Determinants of Chromosome Segregation

描述（由申请人提供）： 项目摘要/摘要 导致染色体不稳定 (CIN) 的突变被认为是导致癌症发生和/或进展的重要诱发事件。我们的方法是开发和应用遗传和生化方法，以了解染色体传递所需的分子成分，总体目标是将我们在酵母中的工作与人类癌症联系起来。具体目标是： 1. 表征重要的 CIN 基因/通路：缩短的端粒 CIN 基因和 ASTRA 复合体。我们将剖析 ASTRA 复合体的功能及其在端粒生物学和 TORC1 信号传导中的作用。 2. 描述 DNA 损伤和 RNA 加工在基因组完整性中的作用。 mRNA 切割和聚腺苷酸化因子 (CPF) 的五个亚基表现出 CIN 和高自发 Rad52 灶发生率，而剪接因子则表现出野生型损伤水平。我们将描述 CPF 复合物和其他 RNA 加工因子导致 DNA 损伤和/或 CIN 的机制。 3. 验证涉及端粒生物学和RNA代谢的CIN基因中的候选体细胞突变，并评估细胞对候选合成致死伙伴的敲低的敏感性。我们将评估酵母中的方法，以确定肿瘤中发现的特定错义体细胞突变是否具有“功能”，并测试从酵母遗传相互作用网络预测的合成致死相互作用，以实现培养哺乳动物细胞中的进化保守性。进一步阐明酵母中CIN的遗传基础将为理解人类细胞中的这一过程提供机制基础，并为癌症中突变的CIN基因提供候选基因。因此，从这项工作中获得的知识将有助于深入了解肿瘤发生。 PHS 398/2590（修订版 06/09） 页面延续 格式页面

DESCRIPTION (provided by applicant): PROJECT SUMMARY/ABSTRACT Mutations that cause chromosome instability (CIN) are considered important predisposing events that contribute to the initiation and/or progression of cancer. Our approach is to develop and apply genetic and biochemical methodologies to obtain an understanding of molecular components required for chromosome transmission, with the overarching goal of relating our work in yeast to human cancer. The specific aims are: 1. To characterize essential CIN genes/pathways: shortened telomere CIN genes and the ASTRA complex. We will dissect the function of the ASTRA complex and its role in telomere biology and TORC1 signaling. 2. To characterize the roles of DNA damage and RNA processing in genome integrity. Five subunits of the mRNA cleavage and polyadenylation factor (CPF) exhibit CIN and high rates of spontaneous Rad52-foci while splicing factors show wild-type levels of damage. We will characterize the mechanism by which the CPF complex and other RNA processing factors cause DNA damage and/or CIN. 3. To validate candidate somatic mutations in CIN genes involved in telomere biology and RNA metabolism, and assess sensitization of cells to knockdown of candidate synthetic lethal partners. We will evaluate methods in yeast to determine whether specific mis-sense somatic mutations found in tumors are "functional" and test synthetic lethal interactions predicted from yeast genetic interaction networks for evolutionary conservation in cultured mammalian cells. Further elucidation of the genetic basis of CIN in yeast will provide a mechanistic basis for understanding this process in human cells, and will provide candidate genes for those CIN genes mutated in cancer. Therefore, knowledge gained from this work will provide insight into tumorigensis. PHS 398/2590 (Rev. 06/09) Page Continuation Format Page