Molecular Dissection of Cytokinesis

细胞分裂的分子解剖

基本信息

批准号：
7240472
负责人：
Michael A Glotzer
金额：
$ 28.44万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-07-01 至 2010-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7240472
关键词：
ATP phosphohydrolase Actomyosin Address Anaphase Animals Binding Biochemical Biochemistry Biological Assay Caenorhabditis elegans Cell Division Process Cell Proliferation Cell Separation Cell division Cells Cellular Structures Coiled-Coil Domain Complex Cytokinesis Defect Development Dissection Ectopic Expression Embryo Excision Factor Analysis Family Genetic Screening Goals In Vitro Kinesin Mammalian Cell Measures Mediating Microtubule Bundle Microtubules Mitotic Mitotic spindle Molecular Molecular Machines Motor Motor Activity Play Property Proteins RNA Interference Reaction Recombinants Relative (related person)Research Research Personnel Role Structure System Tertiary Protein Structure Translating aurora B kinase base cancer therapy daughter cell functional loss in vivo insight member mgcRacGAP novel progenitor programs reconstitution research study rho rho GTPase-activating protein size telophase

项目摘要

DESCRIPTION (provided by applicant): We seek to understand the molecular basis of cell division, cytokinesis, in animal cells. The central spindle, a set of antiparallel bundled microtubules in the anaphase spindle, regulates formation of the actomyosin- based contractile ring and is essential for completion of cytokinesis. We propose that the organization and function of the central spindle is a consequence of the structural organization and biochemical properties of the centralspindlin complex. Centralspindlin is an evolutionary conserved, multimeric complex containing a kinesin-like protein (ZEN-4/MKLP1) and a Rho family GAP (CYK-4/MgcRacGAP) that is highly concentrated on the central spindle and midbody during anaphase and telophase, respectively. We propose to combine in vitro biochemistry and in vivo rescue assays in C. elegans embryos and in mammalian cells to address three specific aims: (1) To dissect the molecular organization of the centralspindlin complex and to characterize the atypical kinesin protein, ZEN-4. By characterizing the critical protein domains in the centralspindlin complex in relative isolation, we will develop the biochemical framework necessary to understand the function of these molecules in more complex reactions as well as in vivo. (2) To decipher the molecular mechanism of central spindle assembly. Central spindle assembly will be reconstituted in vitro from purified components in order to define the principles by which this set of dynamic components assemble into a highly ordered and stable structure. (3) To determine how the central spindle mediates completion of cytokinesis. Centralspindlin contains two distinct protein domains that are proposed to control late steps in cytokinesis. Therefore, the molecular function of these domains will be established. This research will provide molecular insights into a cellular structure that is critical for cell multiplication. Therefore our research could contribute to the development of novel anti-mitotic agents for the treatment of cancer. Furthermore, molecular dissection of ZEN-4 will enhance our understanding of the mechanism of action of microtubule motors.

描述（由申请人提供）：我们试图了解动物细胞中细胞分裂，细胞因子的分子基础。中央主轴是后期纺锤体中的一组反平行捆绑的微管，调节基于肌动蛋白的收缩环的形成，对于完成细胞因子是必不可少的。我们建议中央主轴的组织和功能是中心植物复合体的结构组织和生化特性的结果。 Centralspindlin是一种进化保守的多聚体配合物，其中含有驱动蛋白样蛋白（Zen-4/MKLP1）和Rho家族间隙（CYK-4/MGCRACGAP），分别高度集中在中央纺锤体和中体上的中体和中间体内的中体和固定剂。我们建议在秀丽隐杆线虫胚胎和哺乳动物细胞中结合体外生物化学和体内救援测定法，以解决三个特定目的：（1）剖析Centralspindlin Complect的分子组织，并表征异型蛋白蛋白，Zen-4，Zen-4 。通过在相对分离中表征Centralspindlin复合物中的临界蛋白质结构域，我们将开发出了解这些分子在更复杂的反应以及体内所必需的生化框架。（2）破译中央主轴组件的分子机制。中央主轴组件将在体外从纯化的组件重新结构，以定义这组动态组件组装成高度有序稳定的结构的原理。（3）确定中央主轴如何介导细胞因子的完成。 Centralspindlin包含两个不同的蛋白质结构域，这些结构域建议控制细胞因子的后期步骤。因此，将建立这些结构域的分子功能。这项研究将为细胞结构提供分子见解，这对于细胞繁殖至关重要。因此，我们的研究可能有助于开发新的抗溶毒剂治疗癌症。此外，Zen-4的分子解剖将增强我们对微管电机作用机理的理解。