Regulation of WASH-mediated Endosomal Protein Recycling

WASH 介导的内体蛋白回收的调节

• 批准号: 9353442
• 负责人: Patrick Ryan Potts
• 金额: $ 34.55万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-01 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9353442
• 关键词: Actins; Address; Alzheimer' s Disease; Anthrax disease; Autistic Disorder; Back; Binding; Biochemical; Biological Assay; Carrier Proteins; Cell Line; Cell membrane; Cell physiology; Cells; Cellular biology; Cholera; Complement; Complex; Deubiquitinating Enzyme; Disease; Embryo; Endosomes; Enzymes; Event; F-Actin; Family; Fibroblasts; Generations; Genetic; Golgi Apparatus; Health; Human; Hybrids; In Vitro; Inherited; Kinetics; Knock-out; Knockout Mice; Knowledge; Link; Lysosomes; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Membrane Proteins; Microscopy; Modeling; Molecular; N-terminal; Paralysed; Paraplegia; Pathogenicity; Pathologic; Pathway interactions; Phosphorylation; Phosphorylation Site; Phosphotransferases; Post-Translational Protein Processing; Prader-Willi Syndrome; Process; Property; Protein Cycling Pathway; Protein Sortings; Proteins; Pyrenes; Recombinant Proteins; Recycling; Regulation; Ricin; Role; Signal Pathway; Site; Small Interfering RNA; Testing; Toxin; Transport Process; Ubiquitin; Ubiquitination; Work; base; endosome membrane; human disease; insight; knock-down; member; mutant; prevent; protein complex; protein transport; public health relevance; reconstitution; tool; trans-Golgi Network; ubiquitin-protein ligase

 DESCRIPTION (provided by applicant): Transport of proteins throughout the cell is a highly regulated process that is a fundamental aspect of cell biology. The endosomal protein recycling pathway is one important cellular transport process which promotes the rescue of proteins destined for degradation in the lysosome by recycling them back to the Golgi or plasma membrane from endosomes. A major contributor to this fundamental cellular process is the WASH protein. WASH is part of the WASP family of actin nucleation promoting factors and specifically promotes actin assembly on endosomes to facilitate protein recycling. Importantly, WASH and endosomal protein recycling have been linked to a number of human diseases, including hereditary sporadic paraplegia, Alzheimer's, autism, and cancer. Furthermore, this cellular process is required for the transport of lethal pathogenic toxins, such as cholera, ricin, and anthrax to their site of action. Therefore, understanding the function and regulation of WASH has important implication on human health. WASH is held in an auto-inhibited state due to its incorporation into a multi-subunit complex, referred to as the WASH regulatory complex or SHRC. Recently, we described how WASH is activated by ubiquitination. Non- canonical K63-linked ubiquitination of WASH by the MAGE-L2-TRIM27 E3 ubiquitin ligase is an essential event in WASH activation, generation of endosomal F-actin, and endosomal protein recycling. Utilizing a number of in vitro biochemical and cellular approaches, this project aims to determine the mechanisms and signaling pathways regulating WASH activity through post-translational modifications. These studies are important for our understanding of WASH-mediated endosomal protein recycling, breaking critical barriers to treating human diseases associated with manipulation of this cellular process, and advancing our scientific knowledge of this fundamental cellular process.

 描述（由申请人提供）：蛋白质在整个细胞中的转运是一个高度调节的过程，是细胞生物学的一个基本方面。内体蛋白再循环途径是一种重要的细胞转运过程，其通过将注定在溶酶体中降解的蛋白质从内体再循环回到高尔基体或质膜来促进蛋白质的拯救。这一基本细胞过程的主要贡献者是WASH蛋白。WASH是肌动蛋白成核促进因子的WASP家族的一部分，并且特异性地促进肌动蛋白在内体上组装以促进蛋白质再循环。重要的是，WASH和内体蛋白再循环与许多人类疾病有关，包括遗传性散发性截瘫、阿尔茨海默氏症、自闭症和癌症。此外，这种细胞过程是运输致命的致病毒素所必需的，如霍乱、蓖麻毒素、 和炭疽热的地点因此，了解WASH的功能和调控对人类健康具有重要意义。WASH由于其并入多亚基复合物（称为WASH调节复合物或SHRC）而保持在自抑制状态。最近，我们描述了WASH是如何被泛素化激活的。通过MAGE-L2-TRIM 27 E3泛素连接酶的WASH的非经典K63连接的泛素化是WASH活化、内体F-肌动蛋白的产生和内体蛋白再循环中的重要事件。利用一些体外生物化学和细胞方法，该项目旨在确定通过翻译后修饰调节WASH活性的机制和信号通路。这些研究对于我们理解Wash介导的内体蛋白再循环，打破治疗与操纵该细胞过程相关的人类疾病的关键障碍，以及推进我们对这一基本细胞过程的科学知识非常重要。