The role of CENP-A in mammalian centromere specification

描述（由申请人提供）： 为了使姐妹染色体在细胞分裂期间忠实地分离到相对的子细胞，每条染色体必须具有单个着丝粒。缺乏着丝粒或产生双着丝粒染色体（具有两个着丝粒的染色体）导致有丝分裂期间染色体断裂和染色体错误分离。反过来，异常的染色体分离可能引起发育异常和肿瘤性疾病。此外，更好地了解人类染色体的组成部分，管理遗传信息的传输可能是重要的，在新的基因疗法的发展。具体目标1：通过开发含有内源性小鼠CENP-A基因的条件性破坏的转基因小鼠，这些研究试图确定CENP-A蛋白是否是建立和维持哺乳动物着丝粒所必需的。具体目标二：此外，使用酵母双杂交筛选，本研究致力于确定与CENP-A相互作用的蛋白质，可能参与着丝粒规范和动粒组装。具体目标3：最后，本文提出的研究旨在确定CENP-A本身是否足以招募着丝粒成分并最终指定着丝粒。

DESCRIPTION (provided by applicant): In order for sister chromosomes to be faithfully segregated to opposing daughter cells during cell division, each chromosome must possess a single centromere. The lack of a centromere or the creation of dicentric chromosomes (those possessing two centromeres) leads to chromosomal breakage and missegregation of chromosomes during mitosis. In turn, aberrant chromosome segregation may give rise to developmental abnormalities and neoplastic disease. Furthermore, a better understanding of the components of the human chromosome that govern transmission of genetic information may be important in the development of novel gene therapies. Specific Aim 1: By developing a transgenic mouse containing a conditional disruption of the endogenous mouse CENP-A gene, these studies seek to determine whether CENP-A protein is required for the establishment and maintenance of the mammalian centromere. Specific Aim 2: Furthermore, using a yeast two-hybrid screen, this study endeavors to identify proteins that interact with CENP-A that may be involved in centromere specification and kinetochore assembly. Specific Aim3: Finally, the studies proposed here are designed to determine whether CENP-A itself is sufficient to recruit centromere components and ultimately specify the centromere.