Cell and tumor genetics of mitotic arrest checkpoints

有丝分裂停滞检查点的细胞和肿瘤遗传学

• 批准号: 7054702
• 负责人: PETER Karl SORGER
• 金额: $ 31.55万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-01 至 2006-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7054702
• 关键词: DNA binding protein; RNA interference; aneuploidy; cell cycle; cell growth regulation; chromosome aberrations; cytogenetics; electron microscopy; fibroblasts; fluorescence recovery after photobleaching; fungal genetics; gene targeting; genetic markers; laboratory mouse; neoplasm /cancer genetics; p53 gene /protein; phosphorylation; protein localization; time resolved data; yeasts; DNA binding protein; RNA interference; aneuploidy; cell cycle; cell growth regulation; chromosome aberrations; cytogenetics; electron microscopy; fibroblasts; fluorescence recovery after photobleaching; fungal genetics; gene targeting; genetic markers; laboratory mouse; neoplasm /cancer genetics; p53 gene /protein; phosphorylation; protein localization; time resolved data; yeasts

DESCRIPTION (provided by applicant): The accurate transmission of genetic information during the cell cycle is dependent on the correct operation of a spindle assembly checkpoint the monitors the state of chromosome-microtubule attachment. Signals generated at kinetochores and transmitted via a transduction system comprised of Mad and Bub proteins, act to regulate the activity of the anaphase promoting complex (APC) thereby linking cell cycle progression to the correct execution of the mechanical events of mitosis. The goal of this work is to probe the mechanisms of checkpoint gene activation with the goal of defining the upstream events that initiate checkpoint signaling and the downstream events that control APC. In addition, the role played by checkpoint lesions in genomic instability in mice will be examined through the use of conditional alleles in two checkpoint genes. Specifically: (1) The mechanism by which p53-loss rescues the viability of cells lacking a spindle checkpoint will be analyzed in vitro and in vivo (2) The mechanism of action of a recently discovered negative regulator of Mad2 (CMT2) will be analyzed in detail to uncover key aspects of Mad2-mediated checkpoint signaling. (3) The proteins responsible for recruiting Mad and Bub proteins to kinetochores will be studied, with particular emphasis on the members of the Ndc80 complex (4) Bub1, a kinase with dual functions in checkpoint control and chromosome-microtubule attachment will be analyzed using specific mutations that abrogate various functions (5) The potential role of checkpoint lesions in generating chromosome instability and in promoting cancer will be examined in mice using conditional alleles and mouse model off lung cancer development. The molecular analysis of checkpoints will have an impact on two aspects of cancer biology. First, it should help to reveal the mechanism off action of important anti-microtubule chemotherapeutics such as taxol and the vinca alkaloids. These compounds provoke the spindle checkpoint, and lesions in the checkpoint are very likely to alter the effectiveness of these drugs in the clinic. Second, careful study of the spindle checkpoint should clarify the role of chromosome instability on tumor development.

描述（由申请人提供）：细胞周期中遗传信息的准确传输取决于主轴组件检查站的正确操作，监测染色体 - 微管附件状态。在动力学上产生的信号，并通过由MAD和BUB蛋白组成的转导系统传输，作用于调节后期促进复合物（APC）的活性，从而将细胞周期的进程与有丝分裂的机械事件的正确执行联系起来。这项工作的目的是探测检查点基因激活的机制，目的是定义启动检查点信号传导和控制APC的下游事件的上游事件。此外，检查点病变在小鼠基因组不稳定性中所起的作用将通过在两个检查点基因中使用条件等位基因来检查。具体来说： （1）将在体外和体内分析缺乏主轴检查点的细胞的可行性的机制 （2）将详细分析最近发现的MAD2负面调节剂（CMT2）的作用机理，以发现MAD2介导的检查点信号的关键方面。 （3）将研究负责招募MAD和BUB蛋白到动物学的蛋白质，特别强调NDC80复合体的成员 （4）BUB1，将使用特定的突变分析检查点控制和染色体 - 微管附着的双重功能的激酶，以消除各种功能 （5）检查点病变在产生染色体不稳定性和促进癌症方面的潜在作用将在小鼠中使用条件等位基因和肺癌发展中的小鼠模型检查。 检查点的分子分析将对癌症生物学的两个方面产生影响。首先，它应该有助于揭示重要的抗微管化学治疗剂（例如紫杉醇和VINCA生物碱）的作用的机制。这些化合物引起了主轴检查点，检查点中的病变很可能会改变诊所中这些药物的有效性。其次，对主轴检查点的仔细研究应阐明染色体不稳定性在肿瘤发育中的作用。