ROLE OF THE TUBULIN E-HOOK IN THE ACTIVITY OF MICROTUBULE MOTORS

描述（由申请人提供）：我的长期目标是进行一项研究计划，检查细胞组织的分子基础。我曾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。所有真核生物都利用微管马达如动力蛋白来组织细胞内环境，以配合细胞结构和形态的变化。这些场景包括细胞分裂过程中的基因组划分，蛋白质和细胞器的细胞内运输，以及可能所有形式的细胞迁移。尽管微管马达在这些过程中的核心作用，马达如何与微管基板相互作用，以产生力是知之甚少。 通过本提案中概述的研究，我将开发新的体内和体外系统，以研究微管的结构特征如何影响运动活动。这些工具的开发对于我计划在自己的实验室进行的独立研究项目至关重要。 该提议的广泛目标是检验α-微管蛋白亚基上的带负电荷的E-钩基序通过促进微管马达的结合和/或运动性而有助于微管功能的假设，并确定该基序的作用是否因进化上不同的马达类别而不同。我将特别关注动力蛋白马达，并试图确定动力蛋白与E-钩基序相互作用的分子基础和后果。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，并可能证明对人类疾病的治疗有用。微管马达参与许多疾病，包括神经元病理学和肿瘤发生;因此，了解马达-微管相互作用的分子细节可能导致旨在通过调节特定马达的活性来改变细胞功能的疗法。 公共卫生相关性：这个项目将拓宽我们对微管马达功能的理解，并确定抑制动力蛋白活性的新型药物。这与人类健康有关，因为马达的功能对细胞的组织和功能至关重要。已知运动功能缺陷会导致特定疾病，特别是神经系统疾病，但运动功能障碍如何导致病理的分子基础尚不清楚。此外，电机有助于其他疾病的进展，如病毒感染和肿瘤发生;因此，了解电机功能的分子细节将为开发有效的治疗方法提供信息。

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: This project will broaden our understanding of how microtubule motors function, and identify novel pharmacological agents that inhibit dynein activity. This is relevant to human health because he functions of motora are critical to the organization and function of cells. Defects in motor function are known to cause specific diseases, particularly neurological disorders, but the molecular basis of how motor dysfunction contributes to pathology is not understood. Additionally, motors contribute to the progression of other diseases, such as viral infections and tumorigenesis; therefore, understanding the molecular details of motor function will inform strategies for developing effective therapeutics.