Spatiotemporal Control of Dynein Function by Num1

Num1 对动力蛋白功能的时空控制

• 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
• 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

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）：有丝分裂主轴定位和核迁移对于确定子细胞的大小，位置和发育命运至关重要，并确保忠实的遗传物质隔离。主轴定位是由减去端导向电动机Dynein的活性介导的。动力蛋白是多种基本细胞过程所必需的，包括在细胞分裂过程中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都受到严格的调节。在后期期间，动力蛋白针对并锚定在细胞皮层，其运动活性介导了纺锤体的定位。然而，在酵母中尚不清楚调节动力蛋白活性的机制，已鉴定出皮质锚 - num1-。客观/具体目的：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，我将通过其皮质锚定NUM1检查Dynein的调节。我将在后期检查NUM1 Transit，并确定此过程所需的组件。我将确定NUM1的低聚状态，并确定与NUM1体外相互作用的动力蛋白的亚基。最后，我将确定负责寡聚和动力蛋白相互作用的NUM1域。了解动力蛋白与其皮质锚之间相互作用的确切性质，以及该锚的调节，将取得我们对主轴定位基本生物学过程以及相关人类疾病的理解。