FUNCTIONAL DETERMINANTS OF CHROMOSOME SEGREGATION IN YEAST

染色体不稳定性(CIN)是由癌细胞中的遗传扰动引起的，现已被广泛认识到 是癌症发生和/或进展的主要诱因条件。的总目标是 这项建议是为了鉴定和表征有丝分裂所需的分子成分。 酵母中染色体传递的保真度，并鉴定哺乳动物物种中的同源成分。 具体目标是： 1)。识别CIN表型突变的所有基因，包括必需和非必需基因 功能。三个独立的屏幕将监测染色体标记的不稳定性 用于鉴定对保存基因组很重要的一组全面的蛋白质 稳定性。将对相应的基因组进行注释、交叉检查和比较。 2)。目的：建立人类肿瘤组织中CIN基因突变的遗传互作图谱。我们将生成 酵母中改变的CIN基因集的全面合成致死遗传相互作用图谱 通过基于DNA芯片的筛查发现人类癌症。RNAi将用于验证哺乳动物的表型 细胞。 3)。建立一个系统地交叉参考酵母和人类CIN基因的资源。 与酵母基因同源的哺乳动物基因将通过与酵母的相似性来鉴定 蛋白质查询并存储在带注释的可搜索数据库中。 4)。目的：研究NdClO和动粒总甲基化的生物学功能。突变 将分析NdclO和其他动粒蛋白的废除总甲基化，并将其用作 全基因组筛选的起点 进一步阐明酵母中CIN的遗传基础将为进一步研究CIN提供机制基础。 了解人类细胞的这一过程，将为这些CIN基因提供候选基因 在癌症中发生突变。因此，从这项工作中获得的知识将提供对机制的洞察 关于肿瘤发生的研究。

Chromosome instability (CIN) due to genetic perturbation in cancer cells is now widely recognized to be a major predisposing condition in cancer initiation and/or progression. The general objective of this proposal is to identify and characterize the molecular components required for mitotic chromosome transmission fidelity in yeast and to identify cognate components in mammalian species. The specific aims are: 1). To identify all genes that are mutable to a CIN phenotype, including both essential and nonessential functions. Three independent screens that monitor instability of a chromosomal marker will be used to identify a comprehensive set of proteins that are important for the preservation of genome stability. The corresponding gene sets will be annotated, cross-checked, and compared. 2). To establish genetic interaction maps for CIN genes mutated in human cancer. We will generate comprehensive synthetic lethal genetic interaction maps in yeast for the set of CIN genes altered in human cancer via DNA chip-based screens. RNAi will be used to validate phenotypes in mammalian cells. 3). To establish a resource that systematically cross-references yeast and human CIN genes. Mammalian genes homologous to yeast genes under study will be identified via similarity to yeast protein queries and stored in an annotated searchable database. 4). To investigate the biological functions of NdclO and sumoylation at the kinetochore. Mutations that abrogate sumoylation of NdclO and other kinetochore proteins will be analyzed, and used as starting points for genome-wide screens 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 mechanisms of tumorigenesis.