Role of tubulin CTTs in the activity of microtubule motors

项目摘要 我的长期目标是进行一项研究计划，检查细胞的分子基础， organization.我在华盛顿大学约翰库珀博士的实验室接受博士后培训， 在那里我研究了微管运动动力蛋白。所有的真核生物都使用微管马达，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 方案包括在细胞分裂期间划分基因组，蛋白质的细胞内运输， 细胞器，也许还有各种形式的细胞迁移。尽管微管马达在这些细胞中起着核心作用， 在这一过程中，马达如何与微管基质相互作用以产生力还知之甚少。 通过本提案中概述的研究，我将开发新的体内和体外系统，以研究如何 微管的结构特征影响运动活动。这些工具的开发将是至关重要的， 我计划在我自己的实验室里进行的独立研究项目。 这项提议的主要目的是检验以下假设： α-微管蛋白亚基通过促进微管的结合和/或运动性而有助于微管功能 电机，并确定是否这个主题的作用不同的进化不同类别的电机。我会 特别关注动力蛋白马达，并试图确定动力蛋白的分子基础和后果， 与E钩基序的相互作用。该项目将通过两个目标解决这些问题： 目标1。微管蛋白E-hook在体内是否促进微管马达的活性？ 目标2.在E-钩突变存在下动力蛋白运动的直接分析。 总之，这些分析将提高我们对微管功能的理解， 人类疾病的治疗。微管马达参与许多疾病，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异性抗体的活性来改变细胞功能的疗法。 电动机.

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.