Determining how the spindle assembly checkpoint monitors chromosome biorientation

确定纺锤体装配检查点如何监控染色体生物取向

基本信息

批准号：
8523035
负责人：
David J Wynne
金额：
$ 5.39万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8523035
关键词：
Address Affinity Behavior Binding Biochemical Biochemistry Biological Assay Biology Cell Cycle Cell Cycle Checkpoint Cell Cycle Progression Cell Cycle Regulation Cell division Centromere Chromatin Chromosome Segregation Chromosome Structures Chromosomes Chromosomes, Human, Pair 2 Complex Defect Development Down Syndrome Failure Genome Genomic Instability Goals Health Human Image Imagery Kinetochores Lead Maintenance Malignant Neoplasms Measurement Mediating Mediator of activation protein Microscopic Microscopy Microtubules Mitotic Mitotic Chromosome Mitotic spindle Modeling Monitor Outcome Phosphoric Monoester Hydrolases Phosphorylation Proteins Recombinants Recruitment Activity Regulation Research Research Training Resolution Series Signal Pathway Signal Transduction Sister Sister Chromatid Specificity Stretching System Techniques Technology Testing Training Work Xenopus aurora B kinase cancer cell career drug development egg improved innovation insight meetings member new technology prevent reconstitution research study response tumor progression

项目摘要

DESCRIPTION (provided by applicant): Chromosome segregation must be carefully regulated because missegregation has catastrophic consequences such as genome instability and cancer. In order to segregate properly, all replicated chromosomes must orient correctly on the mitotic spindle with each pair of sister kinetochores making attachments to microtubules from opposing spindle poles. Failure of even a single chromosome to achieve this "bioriented" configuration triggers a major cell cycle checkpoint response, known as the spindle assembly checkpoint (SAC), which halts the cell cycle and prevents separation of sister chromatids. Many of the downstream effectors of the SAC have been well characterized, but the mechanistic details of how misaligned chromosomes trigger the initial activation of the SAC remain poorly understood. A critical player in this regulation is the chromosomal passenger complex (CPC) that controls the activity of the essential mitotic kinase Aurora B. Highlighting the importance of the CPC to cell cycle control, members of CPC are known to be upregulated in cancer cells and are currently being used as targets in drug development. However, despite this importance a thorough understanding of the functions of CPC components is lacking. The research training plan proposed here describes three complementary lines of experimentation to investigate the mechanisms by which the CPC controls the activity of Aurora B and thereby coordinates chromosome structure and cell cycle progression. These studies will provide critical insights into chromosome biology and the regulation of the cell cycle that will have broad relevance to genome maintenance as well as specific impacts on the understanding of cancer progression. The goals of the research training plan proposed here are 1) to determine how the CPC effects the localization and dynamic turnover of Aurora B on mitotic chromosomes 2) to elucidate which targets of Aurora B kinase are effected by chromosome biorientation and 3) to uncover whether the interaction with specific binding partners confers the sensitivity of CPC function to chromosome structure. To meet these goals, three complementary lines of experimentation will be undertaken using the Xenopus egg extract system, which has been widely used to characterize the biochemistry of cell cycle regulation because of the unprecedented control it provides of cell cycle progression. The power of the Xenopus system will be combined with high-resolution microscopic analysis and recent innovations in the reconstitution and purification of mitotic chromatin. These experiments will test current hypotheses for the function of the CPC and will provide valuable training in the application of cutting edge technologies to the important fields of chromosome biology and cell cycle control.

描述（由申请人提供）：必须仔细调节染色体隔离，因为错误分析具有灾难性后果，例如基因组不稳定性和癌症。为了正确隔离，所有复制的染色体都必须在有丝分裂的主轴上正确定向，每对姐妹动物学体从相反的纺锤管上固定在微管上。即使是单个染色体，无法实现这种“生物为导向”的构型的失败会触发主要的细胞周期检查点响应，即纺锤体组装检查点（SAC），该检查点（SAC）停止了细胞周期并防止姐妹染色质被的分离。 SAC的许多下游效应因子都得到了很好的特征，但是染色体未对准的机械细节触发SAC的初始激活仍然鲜为人知。该法规中的关键参与者是控制基本有丝分裂激酶Aurora的活动的染色体乘客综合体（CPC）。 CPC至细胞周期控制，CPC的成员已知在癌细胞中被上调，目前正在用作药物开发的靶标。然而，尽管存在如此重要的是对CPC组件功能的全面了解。这里提出的研究培训计划描述了三种互补的实验线，以研究CPC控制Aurora B活性的机制，从而协调染色体结构和细胞周期进程。这些研究将为染色体生物学和细胞周期的调节提供关键的见解，这将与基因组维持具有广泛相关性以及对癌症进展的理解的特定影响。这里提出的研究培训计划的目标是1）确定CPC如何影响Aurora B对有丝分裂染色体的本地化和动态周转2）阐明Aurora b激酶的靶标通过染色体的生物体现和3）以与特定的结合派对结构相互作用是否与特定的结构结构相互作用。为了实现这些目标，将使用爪蟾卵提取系统进行三种互补的实验线，该系统已被广泛用于表征细胞周期调节的生物化学，因为它提供了细胞周期进展的前所未有的控制。爪蟾系统的功能将与高分辨率显微镜分析以及有丝分裂染色质的重新构造和纯化的最新创新相结合。这些实验将测试当前的假设对CPC的功能，并将在将最尖端技术应用于重要的情况下提供有价值的培训染色体生物学和细胞周期控制的场。