Role of tubulin CTTs in the activity of microtubule motors

微管蛋白 CTT 在微管马达活性中的作用

• 批准号: 8666650
• 负责人: Jeffrey Kyle Moore
• 金额: $ 23.09万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8666650
• 关键词: Address; Affect; Binding; Binding Sites; Biochemical; Biological Assay; Biological Models; Cell Extracts; Cell division; Cell physiology; Cells; Cellular Morphology; Cellular Structures; Cellular biology; Charge; Complement; Defect; Diffusion; Disease; Dynein ATPase; Electrostatics; Environment; Eukaryota; Functional disorder; Genome; Goals; Health; Human; In Vitro; Individual; Investigation; Lead; Length; Microscopy; Microtubules; Molecular; Molecular Genetics; Molecular Motors; Monitor; Motor; Motor Activity; Mutation; Neurons; Organelles; Pathology; Process; Research; Resolution; Role; Saccharomycetales; Source; Surface; System; Testing; Therapeutic; Tubulin; Universities; Virus Diseases; Washington; Work; Yeast Model System; Yeasts; base; cell motility; genetic manipulation; human disease; improved; in vivo; insight; intracellular protein transport; mutant; nervous system disorder; novel; positional cloning; post-doctoral training; programs; protein purification; reconstitution; research study; tool development; tumorigenesis

Project Summary My long-term goal is to conduct a research program that examines the molecular basis of cellular organization. I have received postdoctoral training in the lab of Dr. John Cooper at Washington University, where I have studied the microtubule motor dynein. All eukaryotes employ microtubule motors such as dynein to organize the intracellular environment in coordination with changes in cell structure and morphology. These scenarios include partitioning the genome during cell division, intracellular transport of proteins and organelles, and perhaps all forms of cell migration. Despite the central role for micortubule motors in these processes, how motors interact with the microtubule substrate in order to produce force is poorly understood. With the research outlined in this proposal, I will develop novel in vivo and in vitro systems to examine how structural features on the microtubule influence motor activity. The development of these tools will be critical for the independent research program that I plan to pursue in my own lab. The broad objective of this proposal is to test the hypothesis that the negatively-charged E-hook motif on the a-tubulin subunit contributes to microtubule function by promoting the binding and/or motility of microtubule motors, and determine whether the role of this motif differs for evolutionarily distinct classes of motors. I will particularly focus on the dynein motor, and seek to identify the molecular basis and consequences of dynein's interaction with the E-hook motif. This project will address these issues by pursuing two aims: Aim 1. Does the tubulin E-hook promote the activity of microtubule motors in vivo? Aim 2. Direct analysis of dynein motility in the presence of the E-hook mutations. Together these analyses will improve our understanding of microtubule function, and may proove useful for the treatment of human disease. Microtubule motors are involved in many diseases, including neuronal pathologies and tumorigenesis; therefore, understanding the molecular details of motor-microtubule interactions may lead to therapies aimed at altering cellular function by modulating the activity of specific motors.

项目概要 我的长期目标是开展一项研究计划，检查细胞的分子基础 组织。我在华盛顿大学John Cooper博士的实验室接受了博士后培训， 我在那里研究了微管运动动力蛋白。所有真核生物都使用微管马达，例如动力蛋白 组织细胞内环境与细胞结构和形态的变化相协调。这些 场景包括细胞分裂过程中基因组的划分、蛋白质的细胞内运输以及 细胞器，也许还有所有形式的细胞迁移。尽管微管马达在这些方面发挥着核心作用 对于电机如何与微管基底相互作用以产生力的过程，人们知之甚少。 根据本提案中概述的研究，我将开发新颖的体内和体外系统来研究如何 微管的结构特征影响运动活动。这些工具的开发对于 我计划在自己的实验室中进行的独立研究项目。 该提案的主要目标是测试以下假设：带负电的 E-hook 图案 a-微管蛋白亚基通过促进微管的结合和/或运动来促进微管功能 电机，并确定该基序的作用对于进化上不同的电机类别是否有所不同。我会 特别关注动力蛋白马达，并寻求确定动力蛋白运动的分子基础和后果 与 E-hook 图案的互动。该项目将通过实现两个目标来解决这些问题： 目的 1. 微管蛋白 E-hook 是否促进体内微管马达的活性？ 目标 2. 在 E-hook 突变存在的情况下直接分析动力蛋白运动性。 这些分析共同将提高我们对微管功能的理解，并可能对以下方面有用： 人类疾病的治疗。微管马达与许多疾病有关，包括神经元疾病 病理学和肿瘤发生；因此，了解运动微管的分子细节 相互作用可能导致旨在通过调节特定活性的活性来改变细胞功能的疗法 电机。