INNER CENTROMERE TARGETING OF THE CHROMOSOME PASSENGER COMPLEX

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 有丝分裂在全球水平和局部水平上都受到调控。虽然全球调控推动有丝分裂进程，但局部调控通过在特定时间和地点开启和关闭各种活动来微调有丝分裂。染色体乘客复合体(CPC)是在局部水平上运作的最具特性的有丝分裂调节因子。CPC的定位在整个有丝分裂过程中是动态的，这与其各种有丝分裂活动相对应。在中期前期和中期，CPC定位于着丝粒内侧，纠正微管和着丝粒的错误附着，并产生纺锤体检查点信号。关于CPC如何以及在哪里靶向内着丝粒，我们知之甚少。我建议通过LAP纯化从MNase消化的有丝分裂染色质中纯化CPC，这种方法可以成功地鉴定新的动粒蛋白和着丝粒蛋白，结合质谱学和深度测序来解决这个问题。对识别的蛋白质和DNA的功能的分析将为研究CPC靶向以及内部着丝粒组装奠定坚实的基础。这一提议的长期目标是了解CPC运动如何与有丝分裂进程相耦合，以及这一过程的放松调控如何可能有助于肿瘤的发生。Aurora-B激酶是CPC的核心，已被发现在各种类型的肿瘤中过表达。此外，一类新的针对极光激酶的小分子已被证明是早期临床试验中很有前途的抗癌药物。然而，细胞毒性是一个不可避免的问题，因为这些药物抑制具有多种有丝分裂功能的激酶活性。如果我们能找到一种方法来抑制Aurora-B的特定功能，这个问题可能会被克服。关于CPC目标的知识可能是实现这一目标的有效和必要的方式。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Mitosis is regulated at both global levels and local level. While global regulation drives mitotic progression, the localized regulation fine tunes mitosis by turning on and off various activities at specific times and locations. The chromosome passenger complex (CPC) is the best characterized mitotic regulator that operates at the local level. The localization of the CPC is dynamic throughout mitosis, which corresponds to its various mitotic activities. During premetaphase and metaphase the CPC localizes at inner centromere where it corrects misattachment of microtubule and kinetochore and generates spindle checkpoint signals. Little is known about how and where the CPC is targeted to the inner centromere. I propose to purify CPC from mitotic chromatin digested with MNase by LAP purification, an approach used to successfully identify novel kinetochore proteins and centromere proteins, combined with mass spectrometry and deep sequencing to address this question. Analysis of the function of the identified proteins and DNAs will lay a strong foundation to study CPC targeting as well as inner centromere assembly The long-term goals of this proposal are to understand how CPC movement is coupled to mitotic progression and how deregulation of such a process might contribute to tumorigenesis. Aurora-B kinase, the core of the CPC, has been found to be overexpressed in various types of tumors. In addition, a new class small molecules targeting to Aurora kinases have been proved to be promising anti-cancer drugs in early stage of clinical trials. However, cytotoxicity is an inevitable issue since these drugs inhibit the kinase activity which has a variety of mitotic functions. This issue might be overcome if we can find a way to inhibit a specific function of Aurora-B. Knowledge about CPC targeting might be an efficient and essential way to achieve such a goal.