Inner Centromere Targeting of the Chromosome Passenger Complex

染色体乘客复合物的内部着丝粒靶向

• 批准号: 8111885
• 负责人: Limin Liu
• 金额: $ 5.3万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8111885
• 关键词: Address; Anaphase; Aneuploidy; Antineoplastic Agents; Binding; Biochemical; Biological; Centromere; Chromatin; Chromosome Arm; Chromosomes; Complex; Congenital Abnormality; Coupled; Cytokinesis; DNA Sequence; Foundations; Genetic Materials; Goals; Histone H3; Kinetochores; Knowledge; Location; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Metaphase; Micrococcal Nuclease; Microtubules; Mitosis; Mitotic; Mitotic Activity; Mitotic Chromosome; Movement; Pathway interactions; Pharmaceutical Preparations; Phase I Clinical Trials; Phosphotransferases; Play; Process; Prometaphase; Prophase; Proteins; Regulation; Role; Signal Transduction; Source; Structure; Time; Ubiquitination; aurora B kinase; aurora kinase; cytotoxicity; daughter cell; driving force; novel; overexpression; small molecule; telophase; tumor; tumorigenesis

DESCRIPTION (provided by applicant): 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 [1, 2]. The localization of the CPC is dynamic throughout mitosis, which corresponds to its various mitotic activities [1, 2]. During premetaphase and metaphase the CPC localizes at inner centromere where it corrects misattachment of microtubule and kinetochore [3-10] and generates spindle checkpoint signals [11,12]. 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 [13-15], 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. Meanwhile, it is known TD-60 is required for centromere targeting of the CPC with an unknown mechanism [16]. To determine how TD-60 regualtes CPC targeting, I propose to dissect which structural domain and which biochemical function of TD-60 is involved in CPC regulation through function and structure analysis. 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 [17- 27]. 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 [28]. 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.

描述(由申请人提供)：有丝分裂在全球和地方两个层面上都受到调控。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[1，2]。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[1，2]。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[3-10]，并产生纺锤体检查点信号[11，12]。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[13-15]，并结合质谱学和深度测序来解决这个问题。对鉴定的蛋白质和DNA功能的分析将为研究CPC靶向和内部着丝粒组装奠定坚实的基础。同时，已知TD-60对于着丝粒靶向CPC是必需的，其机制未知[16]。为了确定TD-60如何调节CPC靶向性，我建议通过功能和结构分析来剖析TD-60的哪些结构域和哪些生化功能参与CPC的调节。这项建议的长期目标是了解CPC运动如何与有丝分裂进程相关联，以及放松这一过程可能如何有助于肿瘤的发生。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[17-27]。此外，一类新的针对极光激酶的小分子已被证明在临床试验的早期阶段是很有前途的抗癌药物[28]。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。