Regulation of cytoplasmic dynein in vivo

体内细胞质动力蛋白的调节

• 批准号: 10475621
• 负责人: XIN XIANG
• 金额: $ 38.12万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10475621
• 关键词: Adaptor Signaling Protein; Amyotrophic Lateral Sclerosis; Area; Aspergillus; Aspergillus nidulans; Brain; Cell Survival; Cells; Cessation of life; Collaborations; Complex; Data; Development; Dynein ATPase; Early Endosome; Eukaryotic Cell; Genetic; In Vitro; Kinesin; LIS1 protein; Location; Mental Retardation; Messenger RNA; Microtubules; Minus End of the Microtubule; Molds; Molecular Conformation; Motor; Neurodegenerative Disorders; Neurons; Organelles; Physiological; Plus End of the Microtubule; Process; Proteins; Proteomics; Regulation; Research; Scientist; System; Vesicle; Virion; Virus Diseases; Work; developmental disease; dynactin; experimental study; genome sequencing; human disease; in vivo; insight; lissencephaly; live cell imaging; premature; prevent; therapy development; whole genome

Project Summary/Abstract We study a microtubule motor called cytoplasmic dynein-1 (or “dynein” for simplicity). In eukaryotic cells, microtubules serve as tracks for motor proteins such as dynein and kinesins to move on. These motor proteins deliver cargoes, including organelles, vesicles, proteins, and mRNAs, to different cellular locations for function. A microtubule has two different ends: the plus end facing cell periphery and the minus end close to the cell center. Dynein is a minus-end- directed motor, and it transports cargoes from the cell periphery toward the cell center. Besides the physiological cargoes including early endosomes and other organelles/vesicles, dynein also transports virus particles inward after viral infection. Our lab uses a filamentous fungus called Aspergillus nidulans as a genetic system to study how dynein activities in live cells are controlled by other proteins. We and other scientists have found that dynein gets transported by a kinesin to the microtubule plus end where it interacts with early endosome via adapter proteins such as the HookA complex and the dynactin complex. In live cells, these adapter proteins are required for activating dynein to move toward the microtubule minus end. However, this process also requires other proteins such as LIS1 (Lissencephaly-1) and VezA (a vezatin- like protein). LIS1 promotes an “open” conformation of dynein to facilitate its activation, but the mechanism of VezA is unclear. Our preliminary data also suggest that negative regulators are needed to prevent dynein from moving away from the plus end prematurely without carrying its cargo, but the negative regulators remain to be identified. Moreover, it is unclear what factors regulate cargo release at the microtubule minus end. In the next five years, we will combine classical genetics, live cell imaging with whole genome sequencing to identify both positive and negative regulators of dynein activities. We will also use live cell imaging, proteomics and structural analysis to solve the mechanisms of VezA and newly identified regulators. The Aspergillus genetic system is best suited for these studies. While in vitro experiments are excellent for studying known proteins, our genetic system has the power for discovering unknown regulators. Discovering these regulators will pave the ways leading to new areas of research in the field, which will stimulate further work involving collaborations to gain mechanistic insights into the intricate regulation of the dynein motor. .

项目概要/摘要 我们研究了一种称为细胞质动力蛋白-1（简称为“动力蛋白”）的微管马达。 在真核细胞中，微管充当动力蛋白和驱动蛋白等运动蛋白的轨道， 这些运动蛋白传递货物，包括细胞器、囊泡、蛋白质和 mRNA 到达不同的细胞位置以发挥功能。微管有两个不同的末端：正端。 面向细胞外围的一端和靠近细胞中心的负端是负端。 定向电机，它将货物从细胞外围运输到细胞中心。 生理货物，包括早期内体和其他细胞器/囊泡，还有动力蛋白 病毒感染后将病毒颗粒向内运输我们的实验室使用一种称为丝状真菌。 构巢曲霉作为遗传系统来研究活细胞中动力蛋白的活性 我们和其他科学家发现动力蛋白是由其他蛋白质控制的。 微管正端的驱动蛋白，通过适配器与早期内体相互作用 蛋白质，例如 HookA 复合物和 dynactin 复合物 在活细胞中，这些接头。 激活动力蛋白向微管负端移动需要蛋白质。 这个过程还需要其他蛋白质，例如 LIS1 (Lissencephaly-1) 和 VezA（一种vezatin- LIS1 促进动力蛋白的“开放”构象以促进其激活，但 我们的初步数据还表明，VezA 的机制尚不清楚。 需要防止动力蛋白过早地从正端移开而不携带其 但负面监管因素仍有待确定，而且目前还不清楚是什么因素。 货物调节微管负端的释放在未来五年内，我们将结合起来。 经典遗传学、活细胞成像和全基因组测序来识别阳性和 我们还将使用活细胞成像、蛋白质组学和动力蛋白活性的负调节因子。 结构分析以解决 VezA 和新发现的调节剂的机制。 曲霉遗传系统最适合这些研究。 我们的遗传系统非常适合研究已知蛋白质，有能力发现 发现这些监管机构将为通向新领域铺平道路。 该领域的研究，这将刺激进一步的合作工作，以获得 对动力蛋白电机复杂调节的机械见解。 。