CAREER: Single Molecule Studies to Dissect the Mechanochemistry of Cytoplasmic Dynein

职业：剖析细胞质动力蛋白机械化学的单分子研究

• 批准号: 1055017
• 负责人: Ahmet Yildiz
• 金额: $ 89.26万
• 依托单位: University of California-Berkeley
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1055017&HistoricalAwards=false
• 关键词: CAREER; Single; Molecule; Studies; Dissect

Scientific Merit:Cytoplasmic dynein is a dimeric motor protein responsible for nearly all minus-end directed transport along microtubules in eukaryotic cells. Dynein forms a large dimeric complex and it differs significantly from kinesin motor protein in origin, structure, mechanochemistry and regulation. In comparison with kinesin, whose structure and mechanism are increasingly well understood, we know relatively little about dynein mechanism. The major barriers to studying dynein's motility were the lack of a recombinant expression system and a paucity of tools for observing the movement and conformational states of single motors with sufficient spatial and temporal resolution. The goal of this project is to elucidate the fundamental mechanism of cytoplasmic dynein motility by using state-of-the-art single molecule methods in vitro. Three outstanding questions will be addressed about dynein motility: 1) How is movement of the two motor domains coordinated to maintain high processivity 2) How does ATP hydrolysis in the ring lead to unidirectional motion and which AAA sites are responsible for dynein motility 3) What is the structural basis for dynein's minus-end directionality? Dissecting the mechanism of dynein motility will significantly advance understanding of the way dynein functions in diverse cellular processes.Broader Impacts: This project will also include the following educational and outreach components: 1) Establish an educational outreach program for high school students from underrepresented areas of Oakland, CA. 2) Organize summer research opportunities and science fairs in local public schools to attract young generations to science. 3) Develop a new multidisciplinary graduate level course in biophysical imaging methods. 4) Develop a new curriculum with designated emphasis in Biophysics for Physics majors.

科学价值：细胞质动力蛋白是一种二聚体的马达蛋白，在真核细胞中负责几乎所有沿沿着的负端定向运输。动力蛋白是一种大的二聚体复合物，它与驱动蛋白马达蛋白在起源、结构、机械化学和调控等方面有很大的不同。相对于驱动蛋白的结构和作用机制的日益完善，动力蛋白的作用机制却知之甚少。研究动力蛋白运动性的主要障碍是缺乏重组表达系统和缺乏足够的空间和时间分辨率来观察单个马达的运动和构象状态的工具。本研究的目的是利用最先进的单分子方法在体外研究细胞质动力蛋白运动的基本机制。动力蛋白运动有三个突出的问题：1）两个运动域的运动是如何协调以维持高的持续合成能力的2）环中ATP的水解是如何导致单向运动的以及哪些AAA位点负责动力蛋白的运动3）动力蛋白负端方向性的结构基础是什么？剖析动力蛋白运动的机制将显着推进动力蛋白在不同的细胞processes.Broader影响的功能的方式的理解：该项目还将包括以下教育和推广组件：1）建立一个教育推广计划，高中学生从奥克兰，加利福尼亚州的代表性不足的地区。2)在当地公立学校组织夏季研究机会和科学展览会，吸引年轻一代学习科学。3)开发一个新的多学科研究生水平的生物物理成像方法的课程。4)为物理学专业开发一门以生物物理学为重点的新课程。