Mitotic regulation by checkpoints in budding yeast

芽殖酵母中检查点的有丝分裂调节

基本信息

批准号：
7922751
负责人：
Daniel J Burke
金额：
$ 32.4万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-28 至 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7922751
关键词：
Address Anaphase Aneuploidy Biochemical Markers Biochemistry Cell Cycle Cells Cellular biology Chromosome Segregation Chromosomes Congenital Abnormality DNA Damage Data Development Disease Genes Genetic Genetic Screening Genome Stability Genomic Instability Goals Health Homologous Gene Human Kinetochores Malignant Neoplasms Maps Measures Microtubules Mitosis Mitotic Modeling Monitor Nuclear Pore Complex Pathway interactions Phospho-Specific Antibodies Phosphorylation Process Proteins Recruitment Activity Regulation Regulatory Pathway Research Resolution Role Saccharomyces cerevisiae Saccharomycetales Signal Transduction Sister Chromatid Site Source System Testing driving force gene function improved in vivo insight loss of function member molecular marker mutant novel prevent public health relevance research study response role model segregation transmission process tumor

项目摘要

DESCRIPTION (provided by applicant): Errors in chromosome segregation are the major source of aneuploidy and a driving force in the development of tumors. The spindle assembly checkpoint (SAC) is one of the mechanisms that prevents aneuploidy and is an important regulator of genomic stability. The SAC is orchestrated by the kinetochore and a major unresolved problem, addressed in this proposal, is to determine how the kinetochore measures the lack of tension and lack of occupancy and how the kinetochore transfers that information to the cell cycle machinery. We will complete high resolution mapping of the tension branch of the SAC within the kinetochore using loss-of- function mutants. We have developed a phospho-specific antibody to Mad3 that is the first biochemical marker for the tension branch of the SAC and we propose to use it to dissect this important component of SAC regulation. We propose a new model for the role of the kinetochore in the tension checkpoint. We will determine if subsets of Mad3-containing proteins are phosphorylated, where Mad3 is phosphorylated in the cell and which kinetochore proteins are needed to transmit the tension signal. We have isolated novel mutants that are specific to the occupancy branch of the SAC. We will dissect the role for the SAC in the DNA damage pathway which requires Mec1 and Tel1 phosphorylation of SAC proteins and represents a novel pathway of mitotic regulation. Together, the experiments in this proposal will provide important insights into how SAC signaling is initiated and integrated into the cell cycle. PUBLIC HEALTH RELEVANCE: Errors in chromosome segregation result in imbalances in chromosome number (aneuploidy), a hallmark of some kinds of birth defects and a major driving force in the development of cancer. The spindle assembly checkpoint is one of the mechanisms that prevents aneuploidy and guards against the diseases resulting from genomic instability. The experiments in this proposal will provide important insights into how the spindle checkpoint is organized, how it is regulated and how it is integrated into the cell cycle. The long term goal is to improve human health by preventing aneuploidy that results in birth defects and cancer.

描述（由申请人提供）：染色体分离错误是非整倍性的主要来源，也是肿瘤发展的驱动力。纺锤体组装检查点（SAC）是防止非整倍性的机制之一，也是基因组稳定性的重要调节因子。 SAC 由着丝粒精心策划，本提案中解决的一个主要未解决问题是确定着丝粒如何测量张力的缺乏和占用的缺乏以及着丝粒如何将该信息传输到细胞周期机器。我们将使用功能丧失突变体完成着丝粒内 SAC 张力分支的高分辨率绘图。我们开发了一种针对 Mad3 的磷酸化特异性抗体，它是 SAC 张力分支的第一个生化标记物，我们建议用它来剖析 SAC 调节的这一重要组成部分。我们提出了一个关于着丝粒在张力检查点中的作用的新模型。我们将确定包含 Mad3 的蛋白质子集是否被磷酸化、Mad3 在细胞中的何处被磷酸化以及需要哪些动粒蛋白来传递张力信号。我们分离出了 SAC 占用分支特有的新型突变体。我们将剖析 SAC 在 DNA 损伤途径中的作用，该途径需要 SAC 蛋白的 Mec1 和 Tel1 磷酸化，并代表有丝分裂调节的新途径。总之，本提案中的实验将为 SAC 信号传导如何启动并整合到细胞周期中提供重要的见解。公共卫生相关性：染色体分离错误会导致染色体数量不平衡（非整倍体），这是某些出生缺陷的标志，也是癌症发展的主要驱动力。纺锤体组装检查点是防止非整倍性和预防基因组不稳定引起的疾病的机制之一。该提案中的实验将为纺锤体检查点如何组织、如何调节以及如何整合到细胞周期中提供重要见解。长期目标是通过预防导致出生缺陷和癌症的非整倍体来改善人类健康。