Kinesin Motors and Microtubule-based Trafficking

驱动蛋白马达和基于微管的贩运

• 批准号: 10790194
• 负责人: Kristen J. Verhey
• 金额: $ 2.96万
• 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/10790194
• 关键词: Adopted; Biochemical; Biological; Biophysics; Cell Surface Extensions; 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; 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： 微管对于真核细胞的几乎所有功能都至关重要，从分裂和移动的能力到 它们采用特定形态并承受机械力的能力。微管组装，动力学， 和功能由大量细胞因素决定和调节，包括微管相关 驱动蛋白和动力蛋白超家族中的蛋白质（MAP）和分子马达。我们的总体目标是定义 微管和驱动蛋白驱动哺乳动物细胞内运输的机制 细胞。为此，我们结合了强大的生物物理和生化方法，提供了机制细节 通过细胞检测来研究运动力学和运动性，报告复合体内的调节和功能 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 在我们对细胞内贩运的机制和调节的了解中。我们将定义机制 将蛋白质靶向初级纤毛，这是一种从纤毛表面突出的基于微管的细胞器 细胞并驱动细胞运动和信号传导。我们将利用我们开发的新型化学遗传方法 用于工程可抑制马达，以探测对组装和功能至关重要的驱动蛋白的功能 初级纤毛。我们将使用体外和体外测定驱动蛋白的运动性和产生力的特性。 细胞分析并利用这些知识来理解这些特性是如何通过进化来选择的 细胞中的特定运动功能。最后，我们将通过信号通路测试运动调节模型，例如 刺猬配体。由于微管和驱动蛋白马达的缺陷与发育障碍有关， 神经退行性疾病和癌症，这些研究将增进我们对它们在细胞中功能的理解 生物学和疾病。