Role of RILP in Autophagy

RILP 在自噬中的作用

• 批准号: 10531930
• 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/10531930
• 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; Meditation; Membrane; Microtubules; Modeling; Molecular; Motor; Mutate; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neurodevelopmental Disorder; 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; biophysical analysis; cell motility; developmental disease; dynactin; endosome membrane; genetic regulatory protein; in vivo; inhibition of autophagy; 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; success; 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.

微管马达蛋白负责细胞中的许多运输功能。该提案的重点是 哺乳动物自噬中马达蛋白胞质动力蛋白的一种新机制。自噬是一个关键的 细胞功能，负责回收旧的或受损的蛋白质和细胞器，并清除有毒蛋白质 集料.自噬还涉及神经发育和神经退行性疾病。 细胞质动力蛋白是一种主要的运动蛋白，负责广泛的基本细胞作用，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， 动力蛋白调节剂RILP（Rab-interacting Lysosomal Protein）作为一种新型的神经元和神经细胞的主调节剂， 非神经元自噬我们发现RILP在不同的阶段招募动力蛋白进入自噬体 在整个过程中，通过一系列不同的招募机制， 自噬体蛋白LC 3和ATG 5，以及晚期内体/溶酶体蛋白Rab 7。我们发现RILP 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 我们进一步发现RILP的表达受mTOR激酶控制，mTOR激酶在RILP的表达中起核心作用。 细胞对营养缺乏、损伤和有毒蛋白质聚集的反应。我们进一步发现RILP是 p62（/SQSTM 1）的加工所必需的，在蛋白质清除中的生理作用的直接证据 集料.因此，RILP似乎代表了理解mTOR如何调节细胞凋亡的缺失环节。 细胞机制对不同形式的侮辱或压力的反应。这份建议书是为了制定详细的 RILP调节和功能的机制，特别是在神经元中。目标1将测试mTOR在以下方面的作用： 控制RILP表达，以及PKA在控制RILP/动力蛋白介导的自噬体转运中的作用。目的2 将定义RILP在自噬体生物发生和成熟中的作用。目标3将定义小说的角色 RILP-动力蛋白-动力蛋白-LIS 1超复合物，我们已经分离出调节自噬体运输。的 拟议的研究应该提供重要的洞察到一个基本的新的自噬途径，与基本的 对理解神经退行性疾病和神经发育疾病的病因学和控制的影响。