Spatiotemporal Control of Dynein Function by Num1

Num1 对动力蛋白功能的时空控制

基本信息

批准号：
8419151
负责人：
Elizabeth Sarah Halpin Collins
金额：
$ 5.39万
依托单位：
UNIVERSITY OF MASSACHUSETTS AMHERST
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-01-13 至 2013-01-12
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8419151
关键词：
Affinity Chromatography Anaphase Aneuploidy Animals Behavior Biochemical Biological Assay Biological Process C-terminal Candidate Disease Gene Cell Cycle Cell Polarity Cell division Cell membrane Cell physiology Cells Chromosome Segregation Coiled-Coil Domain Complex Defect Development Distal Dynein ATPase Ensure Exhibits Failure Fluorescence Fluorescence Resonance Energy Transfer Gel Chromatography Genetic Materials Genome Stability Goals Human In Vitro Label Lead Malignant Neoplasms Maps Mediating Membrane Microtubules Mitosis Mitotic spindle Molecular Mothers Motor Motor Activity Movement N-terminal Nature Neck Nuclear Organism PH Domain Pathway interactions Play Population Positioning Attribute Process Proteins Recruitment Activity Regulation Role Saccharomyces cerevisiae Sedimentation process Site Site-Directed Mutagenesis Slide System Testing Yeasts base cell cortex cellular development daughter cell dynactin fungus human disease in vivo migration novel segregation self assembly spatiotemporal

项目摘要

DESCRIPTION (provided by applicant): Mitotic spindle positioning and nuclear migration are critical for determining the size, position and developmental fate of daughter cells, and ensuring faithful segregation of the genetic material. Spindle positioning is mediated by the activity of the minus end-directed motor, dynein. Dynein is required for a variety of fundamental cellular processes, including chromosome segregation and spindle orientation during cell division. Failures of these processes can lead to aneuploidy, which is a hallmark of human cancer. To correctly position the mitotic spindle, dynein function is tightly regulated both spatially and temporally. During anaphase, dynein is targeted to, and anchored at the cell cortex, where its motor activity mediates spindle positioning. The mechanism by which dynein activity is regulated is not known, however, in yeast, a cortical anchor - Num1 - has been identified. OBJECTIVE/SPECIFIC AIMS: My goal is to fully characterize the spatial and temporal regulation of dynein. To answer this fundamental question, I will examine the regulation of dynein by its cortical anchor, Num1. I will examine Num1 transit during anaphase and identify the components required for this process. I will determine oligomeric state of Num1 and identify the subunit of dynein that interacts with Num1 in vitro. Finally, I will determine the Num1 domain responsible for oligomerization and dynein interaction. Understanding the exact nature of the interaction between dynein and its cortical anchor, as well as the regulation of that anchor, will progress our understanding of the basic biological process of spindle positioning, and related human disease.

描述（由申请人提供）：有丝分裂纺锤体定位和核迁移对于确定子细胞的大小、位置和发育命运以及确保遗传物质的忠实分离至关重要。主轴定位是由负端定向电机动力蛋白的活动介导的。动力蛋白是多种基本细胞过程所必需的，包括细胞分裂过程中的染色体分离和纺锤体方向。这些过程的失败可能导致非整倍性，这是人类癌症的标志。为了正确定位有丝分裂纺锤体，动力蛋白功能在空间和时间上受到严格调节。在后期，动力蛋白靶向并锚定在细胞皮层，其运动活动介导纺锤体定位。调节动力蛋白活性的机制尚不清楚，但在酵母中，已鉴定出一个皮质锚 - Num1。目标/具体目标：我的目标是充分表征动力蛋白的空间和时间调节。为了回答这个基本问题，我将研究动力蛋白通过其皮质锚 Num1 的调节。我将在后期检查 Num1 传输并确定此过程所需的组件。我将确定 Num1 的寡聚状态，并鉴定在体外与 Num1 相互作用的动力蛋白亚基。最后，我将确定负责寡聚和动力蛋白相互作用的 Num1 结构域。了解动力蛋白与其皮质锚点之间相互作用的确切性质以及该锚点的调节，将加深我们对纺锤体定位和相关人类疾病的基本生物过程的理解。