Kinesin Motors and Microtubule-based Trafficking

驱动蛋白马达和基于微管的贩运

• 批准号: 10395469
• 负责人: Kristen J. Verhey
• 金额: $ 80.58万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10395469
• 关键词: Adopted; Biochemical; Biological; Biophysics; Cell surface; Cells; Cellular Assay; Cellular biology; Cilia; Complex; Defect; Disease; Dynein ATPase; Engineering; Environment; Erinaceidae; Eukaryotic Cell; Evolution; Goals; In Vitro; Intracellular Transport; Kinesin; Knowledge; Ligands; Link; Malignant Neoplasms; Mammalian Cell; Mechanics; Methods; Microtubule-Associated Proteins; Microtubules; Modeling; Molecular Motors; Morphology; Motor; Mutation; Neurodegenerative Disorders; Organelles; Process; Property; Proteins; Regulation; Reporting; Signal Pathway; Signal Transduction; Testing; Transport Process; Work; base; cell motility; chemical genetics; ciliopathy; developmental disease; genetic approach; human disease; interdisciplinary approach; mechanical force; novel; protein function; trafficking

Project Summary/Abstract: Microtubules are critical for nearly every function of eukaryotic cells, from their ability to divide and move to their ability to adopt specific morphologies and withstand mechanical forces. Microtubule assembly, dyamics, and functions are dictated and regulated by a large number of cellular factors including microtubule associated proteins (MAPs) and molecular motors in the kinesin and dynein superfamilies. Our overall goal is to define the mechanisms by which microtubules and kinesin motor proteins drive intracellular trafficking in mammalian cells. To do this, we combine powerful biophysical and biochemical methods that provide mechanistic detail on motor mechanics and motility with cellular assays that report on regulation and function within the complex cellular environment. We will continue to utilize these multi-disciplinary approaches to investigate critical gaps in our knowledge of mechanisms and regulation of intracellular trafficking. We will define mechanisms for targeting of proteins to the primary cilium, a microtubule-based organelle that protrudes from the surface of the cell and drives cell motility and signaling. We will utilize a novel chemical-genetic approach that we developed for engineering inhibitable motors to probe the functions of kinesins critical for the assembly and function of primary cilia. We will determine the motility and force-generating properties of kinesins using both in vitro and cellular assays and use this knowledge to understand how these properties were selected through evolution for specific motor functions in cells. Finally, we will test models of motor regulation by signaling pathways such as Hedgehog ligand. As defects in microtubules and kinesin motors are linked to developmental disorders, neurodegenerative diseases, and cancer, these studies will advance our understanding of their functions in cell biology and disease.

项目摘要/摘要： 微管对真核细胞的几乎每个功能都至关重要，从它们的分裂和移动的能力 他们采用特定形态并承受机械力的能力。微管组件，dyamics， 和功能由大量细胞因子（包括微管相关）指示和调节 蛋白质（地图）和分子电动机和动力蛋白超家族中的分子电机。我们的总体目标是定义 微管和驱动蛋白运动蛋白在哺乳动物中驱动细胞内运输的机制 细胞。为此，我们结合了强大的生物物理和生化方法，以提供机械细节 关于运动力学和运动性的细胞测定，报告了有关调节和功能的调节 细胞环境。我们将继续利用这些多学科方法来调查关键差距 在我们了解机制和细胞内贩运的调节中。我们将定义机制 将蛋白质靶向原发性纤毛，这是一种基于微管的细胞器，从表面突出 细胞并驱动细胞的运动性和信号传导。我们将利用一种新型的化学遗传方法 用于工程可抑制的电动机，以探测驱动蛋白的功能 原发性纤毛。我们将使用体外和 细胞测定并利用这些知识来了解如何通过进化选择这些特性 细胞中的特定运动功能。最后，我们将通过信号通路（例如 刺猬配体。由于微管和动力素电机的缺陷与发育障碍有关 神经退行性疾病和癌症，这些研究将提高我们对细胞功能的理解 生物学和疾病。