INNER CENTROMERE TARGETING OF THE CHROMOSOME PASSENGER COMPLEX

染色体过客复合体的内着丝粒靶向

• 批准号: 8365783
• 负责人: P. TODD STUKENBERG
• 金额: $ 1.28万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365783
• 关键词: Address; Antineoplastic Agents; Biology; Centromere; Chromatin; Chromosomes; Complex; Coupled; Foundations; Funding; Fungal Genome; Goals; Grant; Kinetochores; Knowledge; Location; Mass Spectrum Analysis; Metaphase; Microtubules; Mitosis; Mitotic; Mitotic Activity; Movement; National Center for Research Resources; Pharmaceutical Preparations; Phase I Clinical Trials; Phosphotransferases; Principal Investigator; Process; Proteins; Regulation; Research; Research Infrastructure; Resources; Signal Transduction; Source; Time; United States National Institutes of Health; aurora B kinase; aurora kinase; cost; cytotoxicity; novel; overexpression; small molecule; tumor; tumorigenesis

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.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 有丝分裂在全球和地方两个层面都受到调控。虽然全局调节驱动有丝分裂进程，但局部调节通过在特定时间和位置开启和关闭各种活动来微调有丝分裂。染色体乘客复合物（CPC）是最具特征的有丝分裂调节剂，在当地的水平上运作。CPC的定位在整个有丝分裂中是动态的，这对应于其各种有丝分裂活动。在前中期和中期，CPC定位于内着丝粒，在那里它纠正微管和动粒的错误附着，并产生纺锤体检查点信号。关于CPC如何以及在何处被靶向到内部着丝粒，知之甚少。我建议从MNase消化的有丝分裂染色质中纯化CPC，这种方法用于成功地识别新的动粒蛋白和着丝粒蛋白，结合质谱和深度测序来解决这个问题。分析所鉴定的蛋白质和DNA的功能将为研究CPC靶向以及内部着丝粒组装奠定坚实的基础。本提案的长期目标是了解CPC运动如何与有丝分裂进程相耦合以及这种过程的失调如何有助于肿瘤发生。已发现CPC的核心Aurora-B激酶在各种类型的肿瘤中过表达。另外，一类新的靶向Aurora激酶的小分子药物已在早期临床试验中被证明是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，因为这些药物抑制激酶的活性，具有多种有丝分裂功能。如果我们能找到一种抑制Aurora-B特定功能的方法，这个问题可能会得到解决。了解产品总分类的目标可能是实现这一目标的一个有效和必要的途径。