Mechanisms of spindle formation

纺锤体形成机制

基本信息

批准号：
8546426
负责人：
PHONG T TRAN
金额：
$ 32.19万
依托单位：
UNIVERSITY OF PENNSYLVANIA
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：
2012-09-30 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8546426
关键词：
Address Anaphase Aneuploidy Binding Biochemical Biochemistry Biological Assay Biological Models COX7A2L Protein Cell Cycle Cell division Cells Cellular biology Centrosome Chromosome Segregation Chromosomes Cultured Cells Defect Dependency Dynein ATPase Eukaryotic Cell Event Failure Fission Yeast Frequencies Genes Genetic Hela Cells Homologous Gene Human Kinesin Kinetochores Laboratories Lead Learning Length Life Macromolecular Complexes Malignant Neoplasms Metaphase Microfluidic Microchips Microfluidics Microtubule-Associated Proteins Microtubules Mitosis Mitotic spindle Modeling Molecular Molecular Biology Techniques Molecular Motors Motor Mutagenesis Names Nuclear Envelope Optics Phase Phase Transition Phenotype Phosphoric Monoester Hydrolases Phosphotransferases Play Plus End of the Microtubule Prophase Proteins Resolution Role Slide Small Interfering RNA Staging Structure Techniques Tertiary Protein Structure Testing Work cancer type cell cortex cellular imaging crosslink daughter cell gene discovery genetic regulatory protein innovation interest novel polymerization segregation spindle pole body telophase tool

项目摘要

DESCRIPTION (provided by applicant): Mitosis is a key stage during the life of a cell. It is the stage where a bipolar spindle structure is organized to segregate duplicated chromosomes into the two daughter cells. Spindle organization and function require exquisite precision, robustness and fidelity. Defects associated with the spindle can lead to defects in chromosomal segregation, or aneuploidy, which has been correlated with some types of cancer. The spindle is a macromolecular machine made of microtubules, microtubule-associated proteins (MAPs), molecular motors and other regulatory proteins. Of intense interest have been molecular motors, which perform work such as cross-linking and sliding microtubules apart to form the bipolar spindle, or to depolymerize microtubules to maintain proper spindle lengths, or to carry chromosomes to opposite spindle poles. Surprisingly, while we have learned much about motors involved in mitosis, we still know very little about the MAPs and other regulatory proteins and how they coordinate with motors to bring about proper spindle formation. My laboratory uses the relatively simple fission yeast Schizosaccharomyces pombe and human cultured cells to address conserved mechanisms of spindle organization and function. This particular project focuses on how the initial bipolar spindle is formed at the start of mitosis, the stage termed prophase. We focus on the MAPs that contribute to spindle formation. Using fission yeast as a gene discovery tool, we have begun to define the roles of a new gene we called psr1+ (poles separation regulator 1). Our work indicates that psr1p organizes the initial bipolar spindle during prophase. Psr1-deletion leads to high frequency of monopolar spindles and subsequent chromosome segregation defects. Fission yeast psr1+ appears to have a human functional homolog. We have begun to characterize a novel human gene we called PSR1. In HeLa cells, siRNA of PSR1 also leads to high frequency of monopolar spindles and subsequent chromosome segregation defects. This proposal aims to combine modern cell and molecular biology techniques in fission yeast and human cultured cells, biochemistry, high-resolution optical live-cell imaging, and innovative microfluidic techniques to control cellular microenvironment, to reach a mechanistic understanding of bipolar spindle formation.

描述（由申请人提供）：有丝分裂是细胞寿命期间的关键阶段。这是组织双极主轴结构将重复的染色体隔离到两个子细胞中的阶段。主轴组织和功能需要精确的精度，鲁棒性和忠诚度。与纺锤体相关的缺陷会导致与某些类型的癌症相关的染色体分离或非整倍性的缺陷。纺锤体是由微管，微管相关蛋白（地图），分子电动机和其他调节蛋白制成的大分子机器。非常感兴趣的是分子电动机，它们进行的工作，例如交联和滑动微管以形成双极纺锤体，或者将微管解散以保持适当的纺锤体长度，或将染色体携带到相对的纺锤体杆上。令人惊讶的是，尽管我们对涉及有丝分裂的电机有了很多了解，但我们仍然对地图和其他调节蛋白以及它们如何与电动机进行协调以实现适当的纺锤体形成。我的实验室使用相对简单的裂变酵母菌菌果糖和人类培养细胞来解决纺锤体组织和功能的保守机制。这个特殊的项目着重于在有丝分裂开始时如何形成初始双极纺锤体，该阶段称为预言。我们专注于有助于主轴形成的地图。使用裂变酵母作为基因发现工具，我们开始定义称为PSR1+的新基因的作用（杆分离调节剂1）。我们的工作表明PSR1P在前期。 PSR1删除导致单极纺锤体的高频和随后的染色体隔离缺陷。裂变酵母PSR1+似乎具有人类功能同源物。我们已经开始描述一个称为PSR1的新型人类基因。在HeLa细胞中，PSR1的siRNA还导致单极纺锤体的高频和随后的染色体隔离缺陷。该提案旨在将现代细胞和分子生物学技术结合在裂变酵母和人培养细胞中，生物化学，高分辨率光学活细胞成像以及创新的微流体技术来控制细胞微环境，以达到对双极链球形成的机械理解。