Regulation of the MAVS Signalosome by RNA and RNA-binding Proteins

RNA 和 RNA 结合蛋白对 MAVS 信号体的调节

• 批准号: 10640526
• 负责人: Nandan Satish Gokhale
• 金额: $ 12.79万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10640526
• 关键词: Adaptor Signaling Protein; Advisory Committees; Autoimmune Diseases; Biochemical; Bioinformatics; Biological Models; Co-Immunoprecipitations; Complex; Coupled; Cytoplasmic Protein; Cytosol; Data; Elements; Environment; Family; Fractionation; Future; Gene Expression; Goals; Immune; Immune signaling; Immunologic Stimulation; Infection; Infection prevention; Inflammation; Inflammatory; Interferons; Mass Spectrum Analysis; Mediating; Membrane; Mentors; Microscopy; Mitochondria; Molecular; Natural Immunity; Optics; Organelles; Outcome; Pathway interactions; Pattern recognition receptor; Phase; Post-Translational Protein Processing; Process; Production; Proteins; Proteomics; RNA; RNA Binding; RNA Viruses; RNA-Binding Proteins; RNA-Protein Interaction; Receptor Signaling; Regulation; Research; Research Personnel; Ribonucleases; Ribosomes; Role; Scientist; Signal Pathway; Signal Transduction; Site; Spliceosomes; Structure; Sucrose; TBK1 gene; TNF receptor-associated factor 3; TRAF2 gene; Techniques; Testing; Therapeutic; Tissues; Training; Universities; Viral; Virus Diseases; Washington; Work; adaptive immune response; antiviral immunity; career; combat; druggable target; experience; experimental study; immune activation; novel; novel therapeutic intervention; pathogenic virus; prevent; programs; protein complex; protein protein interaction; receptor; response; scaffold; screening; skills; viral RNA; virology

PROJECT SUMMARY / ABSTRACT Well controlled antiviral innate immunity is essential for restricting viral pathogens while preventing aberrant inflammation. A key antiviral signaling pathway is initiated by RIG-I-like receptors (RLRs) which sense viral RNA in the cytosol to induce the production of interferons through a higher order signaling platform centered on the adaptor protein MAVS. Although the components of the MAVS signalosome are basally expressed, RLR signaling only proceeds when they coalesce around MAVS at ER-mitochondrial contact sites. Many of the protein-protein interactions and post-translational modifications required for the proper function of the MAVS signalosome are known. While, functional interactions in protein complexes can be mediated by RNA molecules, whether RNA molecules play a role in signaling through the MAVS signalosome remains unexplored. I have found that 1) MAVS is associated with non-viral RNA, that 2) RNA promotes signaling through the MAVS signalosome, and that 3) canonical RNA-binding proteins (RBPs) interact with MAVS. This proposal aims to define the functional roles for RNA and RBPs in regulating antiviral signaling through the MAVS signalosome. During the mentored phase, I will gain new training in the characterization of RNA-protein interactions through biochemical, sequencing-based, and quantitative proteomic approaches, as well as the virological techniques required to study RNA viruses and innate immunity. In Aim 1, I will define the RNA-dependent interactions between components of the MAVS signalosome during the K99 phase, and identify novel proteins that associate with RNA during RLR signaling at the R00 phase. Through Aim 2, I will pinpoint the sequences in MAVS required for RNA-association, and identify a high-confidence set of MAVS-bound RNAs during the K99 phase. During the R00 stage, I will functionally dissect the roles for these RNAs at the MAVS signalosome. In Aim 3, I will screen a shortlist of MAVS-associated RBPs to identify those that influence MAVS signaling during the K99 stage, and determine the molecular functions of three RBPs (STAU1, HNRNPL, and RBM10) in antiviral signaling at the R00 stage. The overall outcome of these experiments will be to define new RNA-centric principles by which the MAVS signalosome is organized. Understanding how RNA molecules influence antiviral signaling could unlock new host-directed therapeutic strategies against viral diseases as well as autoimmune disorders. In addition to my advisor Dr. Savan, I have assembled an Advisory Committee with expertise in the different facets of my research. Together, this excellent training environment at the University of Washington will augment my research during the mentored phase, and equip me with the skills required to transition to an independent academic researcher studying the RNA regulation of innate immune processes.

项目概要/摘要 良好控制的抗病毒先天免疫对于限制病毒病原体同时防止异常至关重要 炎。关键的抗病毒信号传导途径由 RIG-I 样受体 (RLR) 启动，该受体可感知病毒 RNA 在细胞质中通过以 接头蛋白 MAVS。尽管 MAVS 信号体的成分是基础表达的，但 RLR 仅当它们在 ER 线粒体接触位点围绕 MAVS 结合时，信号传导才会发生。许多 MAVS 正常功能所需的蛋白质-蛋白质相互作用和翻译后修饰 信号体是已知的。同时，蛋白质复合物中的功能相互作用可以由 RNA 分子介导， RNA 分子是否在通过 MAVS 信号体的信号传导中发挥作用仍有待探索。我有 发现 1) MAVS 与非病毒 RNA 相关，2) RNA 通过 MAVS 促进信号传导 信号体，以及 3) 经典 RNA 结合蛋白 (RBP) 与 MAVS 相互作用。该提案旨在 定义了 RNA 和 RBP 在通过 MAVS 信号体调节抗病毒信号传导中的功能作用。 在指导阶段，我将通过以下方式获得有关 RNA-蛋白质相互作用表征的新培训： 生物化学、基于测序和定量蛋白质组学方法以及病毒学技术 研究 RNA 病毒和先天免疫所需。在目标 1 中，我将定义 RNA 依赖性相互作用 K99 阶段 MAVS 信号体成分之间的关​​系，并鉴定出关联的新蛋白质 在 R00 期的 RLR 信号传导过程中与 RNA 结合。通过目标 2，我将查明 MAVS 中所需的序列 用于 RNA 关联，并在 K99 阶段鉴定一组高置信度的 MAVS 结合 RNA。期间 R00 阶段，我将从功能上剖析这些 RNA 在 MAVS 信号体中的作用。在目标 3 中，我将筛选 MAVS 相关 RBP 的候选名单，用于识别那些在 K99 阶段影响 MAVS 信号传导的 RBP，以​​及 确定三个 RBP（STAU1、HNRNPL 和 RBM10）在抗病毒信号传导中的分子功能 R00阶段。这些实验的总体结果将是定义新的以 RNA 为中心的原则，通过该原则 MAVS 信号体是有组织的。了解 RNA 分子如何影响抗病毒信号传导可以解开谜团 针对病毒性疾病和自身免疫性疾病的新的宿主导向治疗策略。此外 我的顾问萨万博士，我组建了一个咨询委员会，在我的项目的不同方面拥有专业知识 研究。总之，华盛顿大学良好的培训环境将增强我的研究成果 在指导阶段，让我具备过渡到独立学者所需的技能 研究人员研究先天免疫过程的 RNA 调节。