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.

项目摘要 我的长期目标是进行研究计划，以检查细胞的分子基础 组织。我已经在华盛顿大学约翰·库珀博士的实验室接受了博士后培训， 我研究了微管运动蛋白的地方。所有真核生物都采用微管电机，例如动力蛋白 与细胞结构和形态变化的协调中组织细胞内环境。这些 场景包括在细胞分裂期间分区基因组，蛋白质的细胞内转运和 细胞器，也许是所有形式的细胞迁移。尽管在这些方 流程，电动机如何与微管底物相互作用以产生力的情况很少了解。 通过此提案中概述的研究，我将在体内和体外系统中开发新颖的，以研究如何 微管上的结构特征会影响运动活动。这些工具的开发对于 我打算在自己的实验室中进行的独立研究计划。 该提议的广泛目的是检验以下假设： A-微管蛋白亚基通过促进微管的结合和/或运动有助于微管功能 电动机，并确定该基序的作用在进化不同的电动机上是否有所不同。我会 特别专注于动力蛋白电动机，并寻求识别Dynein的分子基础和后果 与电子钩基序相互作用。该项目将通过追求两个目标来解决这些问题： AIM 1。小管蛋白电子钩是否会促进体内微管电动机的活性？ 目标2。在存在电子钩突变的情况下直接分析动力蛋白运动。 这些分析一起将提高我们对微管功能的理解，并可能对 人类疾病的治疗。微管电机参与了许多疾病，包括神经元 病理和肿瘤发生；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来改变细胞功能的疗法 电动机。