Molecular Dissection of the "Pacman-Flux" Machinery Used to Move Chromosomes

用于移动染色体的“Pacman-Flux”机器的分子解剖

基本信息

批准号：
8000101
负责人：
DAVID James SHARP
金额：
$ 10.05万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-01-14 至 2011-01-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): The ability of chromosomes to move towards opposite poles of the mitotic spindle is fundamental for proper cell division and defects in this process are believed to be an initial step in tumorigenesis. Poleward chromosome motility occurs by a "Pacman-flux" mechanism: Chromosomes induce the depolymerization of attached microtubule plus-ends, termed "Pacman", while being reeled into spindle poles by poleward tubulin flux" stimulated by depolymerization of microtubule minus-ends. The goal of the studies outlined in this proposal is to elucidate the molecular machinery that stimulates and controls "Pacman-flux"-based chromosome motility. The central hypothesis of this work is that chromosome motility is controlled by a mechanistically diverse array of proteins which employ distinct targeting and mechanisms-of-action to control the polymerization state of microtubule ends. There are two specific aims: Aim 1) Elucidate the molecular pathway that stimulates and controls the velocity of microtubule minus-end depolymerization and poleward flux. Studies in this aim evaluate the flux-related functions of microtubule severing proteins, multiple kinesin-13s and kinesin-13 phosphorylation. Aim 2) Elucidate the molecular pathway by which chromosomes induce the depolymerization of microtubule plus-ends. Studies in this aim evaluate the Pacman-related functions of CLIP-170/190, microtubule severing proteins, and multiple kinesin-13s. The fruit fly Drosophila melanogaster is the primary experimental system used in these studies. Live cell techniques for visualizing spindle and chromosome dynamics have been optimized in Drosophila S2 cells and embryos and thus these cells provide an ideal context within which to study how the manipulation of protein function impacts "Pacman-Flux". Live cell microscopic studies of mitosis will be complemented by biochemical and molecular approaches to provide an in-depth understanding of whether and how specific classes of proteins drive chromosome segregation. Additional studies are performed in human cells to determine whether aspects of our proposed pathways are evolutionary conserved. Defects in chromosome segregation lead to human maladies such as birth defects and cancer. An understanding of how this process occurs normally should provide insights into the molecular etiology of these diseases and suggest therapeutic strategies for their treatment.

描述（由申请人提供）：染色体向相反的有丝分裂纺锤体移动的能力对于适当的细胞分裂至关重要，并且在此过程中的缺陷被认为是肿瘤发生的第一步。极向染色体运动通过“ PACMAN-流量”机制发生：染色体诱导附着的微管加末端的沉降，称为“ PACMAN”，同时被极点小管的旋转量刺激了摩尔元素的摩尔元素，从而刺激了极线细胞的刺激。刺激和控制基于“ PACMAN-FLUX”的染色体运动的中心假设。微管末端的末端解放和极化的速度。该目标的研究评估了夹170/190，微管切断蛋白和多个驱动蛋白13的与PACMAN相关的功能。果蝇果蝇Melanogaster是这些研究中使用的主要实验系统。可视化纺锤体和染色体动力学的活细胞技术已在果蝇S2细胞和胚胎中进行了优化，因此这些细胞提供了一个理想的环境，以研究蛋白质功能的操纵如何影响“ Pacman-Flux”。生物化学和分子方法将补充有丝分裂的活细胞显微镜研究，以深入了解特定类别的蛋白质是否以及如何驱动染色体隔离。在人类细胞中进行了其他研究，以确定我们提出的途径的各个方面是否是进化的。染色体分离的缺陷导致人类疾病，例如先天缺陷和癌症。对这个过程通常如何发生的理解应提供对这些疾病的分子病因的见解，并提出治疗治疗的治疗策略。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Drosophila katanin is a microtubule depolymerase that regulates cortical-microtubule plus-end interactions and cell migration.

DOI：
10.1038/ncb2206
发表时间：
2011-04
期刊：
Nature cell biology
影响因子：
21.3
作者：
Zhang D;Grode KD;Stewman SF;Diaz-Valencia JD;Liebling E;Rath U;Riera T;Currie JD;Buster DW;Asenjo AB;Sosa HJ;Ross JL;Ma A;Rogers SL;Sharp DJ
通讯作者：
Sharp DJ

Motor domain phosphorylation and regulation of the Drosophila kinesin 13, KLP10A.