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，研究细胞的分子基础 组织。我在华盛顿大学约翰·库珀博士的实验室接受了博士后培训， 在那里我研究了微管运动动力蛋白。所有真核生物都利用动力蛋白等微管马达 根据细胞结构和形态的变化来组织细胞内的环境。这些 情景包括在细胞分裂期间分割基因组，蛋白质在细胞内的运输，以及 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方面发挥了核心作用 在这一过程中，马达如何与微管底物相互作用以产生力还知之甚少。 根据这项提案中概述的研究，我将开发新的体内和体外系统，以检查如何 微管的结构特征影响运动活动。这些工具的开发将对 我计划在自己的实验室里进行的独立研究项目。 这一提议的广泛目标是检验这样的假设，即带负电荷的E-挂钩基序在 微管蛋白A亚基通过促进微管的结合和/或运动来促进微管的功能 马达，并确定这个基序的作用是否不同于进化上不同的马达类别。这就做 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-Hook主题的相互作用。该项目将通过追求两个目标来解决这些问题： 目的1.微管蛋白E挂钩在体内是否促进微管马达的活性？ 目的2.直接分析存在E-挂钩突变时的动力蛋白运动性。 总之，这些分析将提高我们对微管功能的理解，并可能证明 人类疾病的治疗。微管马达与许多疾病有关，包括神经元。 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的细胞活性来改变细胞功能的治疗 摩托公司。

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.