Role of RILP in Autophagy

RILP 在自噬中的作用

• 批准号: 10307619
• 负责人: Richard Bert Vallee
• 金额: $ 35.73万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10307619
• 关键词: AGFG1 gene; Adaptor Signaling Protein; Autophagocytosis; Autophagosome; Axonal Transport; Behavior; Behavioral Mechanisms; Binding; Binding Proteins; Binding Sites; Biogenesis; Biological; Biophysical Process; Cell division; Cell physiology; Cells; Cellular Stress; Collaborations; Cyclic AMP-Dependent Protein Kinases; Cytoplasm; Cytoplasmic Protein; Data; Defect; Defense Mechanisms; Disease; Docking; Dynein ATPase; Endosomes; Etiology; FRAP1 gene; Genes; Image; In Vitro; Injury; Intracellular Transport; Link; Lysosomes; Mediating; Membrane; Microtubules; Modeling; Molecular; Motor; Mutate; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neuroglia; Neurologic; Neurons; Nuclear; Organelles; Pathway interactions; Peptides; Phosphotransferases; Physiological; Play; Process; Production; Promoter Regions; Protein Analysis; Proteins; Recycling; Regulation; Role; Signal Transduction; Site; Starvation; Stress; Testing; Work; base; biophysical analysis; cell motility; developmental disease; dynactin; genetic regulatory protein; in vivo; inhibitor; insight; interest; knock-down; late endosome; lysosomal proteins; mTOR inhibition; neurodevelopment; novel; nutrient deprivation; pathogen; protein activation; protein aggregation; protein complex; protein expression; protein function; protein protein interaction; rab7 protein; recruit; response; single molecule; small molecule; transcription factor

Microtubule motor proteins are responsible for numerous transport functions in cells. This proposal focuses on a novel mechanism for the motor protein cytoplasmic dynein in mammalian autophagy. Autophagy is a critical cellular function responsible for recycling old or damaged proteins and organelles, and for clearing toxic protein aggregates. Autophagy is also implicated in neurodevelopmental and neurodegenerative diseases. Cytoplasmic dynein is a major motor protein responsible for a broad range of basic cellular roles, including retrograde axonal transport, cell division, and nuclear and cell migration. We have now found that the cytoplasmic dynein regulator, RILP (Rab-interacting Lysosomal Protein) acts as a novel master regulator of neuronal and nonneuronal autophagy. We find that RILP recruits dynein to autophagosomes at a succesion of stages throughout this process via a sequence of distinct recruitment mechanisms involving interactions with the autophagosomal proteins LC3 and ATG5, as well as the late endosomal/lysosomal protein Rab7. We find RILP mediates not only autophagosome transport, but has a surprising role in autophagosome biogenesis as well. Of further interest we find RILP expression to be controlled by the mTOR kinase, which plays a central role in the cellular response to nutrient deprivation, injury, and toxic protein aggregation. We find further that RILP is necessary for processing of p62(/SQSTM1), direct evidence for a physiological role in clearance of protein aggregates. RILP appears, therefore, to represent a missing link in understanding how mTOR regulates the cellular machinery in response to diverse forms of insult or stress. This proposal is to work out the detailed mechanisms for RILP regulation and function, especially in neurons. Aim 1 will test the role of mTOR in controlling RILP expression, and of PKA in controlling RILP/dynein-mediated autophagosome transport. Aim 2 Will define the roles of RILP in autophagosome biogenesis and maturation. Aim 3 will define the role of a novel RILP-dynein-dynactin-LIS1 supercomplex we have isolated in regulating autophagosome transport. The proposed studies should provide important insight into a basic new autophagy pathway, with fundamental implications for understanding the etiology and control of neurodegenerative and neurodevelopmental diseases.

微管运动蛋白负责细胞中的许多运输功能。该提议重点 哺乳动物自噬中运动蛋白细胞质动力蛋白的一种新型机制。自噬是关键 负责回收旧或受损蛋白质和细胞器的细胞功能，以及清除有毒蛋白 聚合。自噬也与神经发育和神经退行性疾病有关。 细胞质动力蛋白是负责广泛基本细胞角色的主要运动蛋白，包括 逆行轴突运输，细胞分裂以及核和细胞迁移。我们现在发现细胞质 Dynein调节剂RILP（Rab Interacting溶酶体蛋白）充当神经元和神经元的新主调节剂 非神经元自噬。我们发现RILP在阶段的成功中将动力蛋白招募到自噬体 在整个过程中，通过一系列不同的招募机制，涉及与 自噬体蛋白LC3和ATG5，以及晚期内体/溶酶体蛋白Rab7。我们找到了里尔 不仅介导自噬体转运，而且在自噬体生物发生中也具有令人惊讶的作用。的 进一步的兴趣，我们发现RILP表达受MTOR激酶的控制，该激酶在 细胞对营养剥夺，损伤和有毒蛋白质聚集的反应。我们进一步发现里尔普是 处理p62（/sqstm1）所需的直接证据证明生理作用在清除蛋白质中 聚合。因此，RILP似乎是在理解MTOR如何调节的情况下表示缺失的链接 蜂窝机械响应各种形式的侮辱或压力。该提议是为了解决详细的 RILP调节和功能的机制，尤其是在神经元中。 AIM 1将测试MTOR在 控制RILP/Dynein介导的自噬体转运中的RILP表达和PKA。目标2 将定义RILP在自噬体生物发生和成熟中的作用。 AIM 3将定义小说的作用 rilp-dynein-dynactin-lis1超复合物我们在调节自噬体转运方面分离出来。这 拟议的研究应提供对基本新自噬途径的重要见解，并具有基本 理解神经退行性和神经发育疾病的病因和控制的意义。