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.

项目摘要 /摘要 控制良好的抗病毒先天免疫对于限制病毒病原体至关重要 炎。关键的抗病毒信号通路是由RIG-I样受体（RLR）启动的，该受体感知病毒RNA 在细胞质中，通过以上为中心的高阶信号平台诱导干扰素的产生 适配器蛋白MAV。尽管MAVS信号体的成分基本表达，但RLR 只有在ER线粒体接触位点围绕MAV结合的MAV时，信号才会进行。许多 蛋白质 - 蛋白质相互作用和MAV适当功能所需的翻译后修饰 信号体是已知的。而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信号体信号传导中发挥作用。我有 发现1）MAV与非病毒RNA有关，2）RNA通过MAV促进信号传导 信号体和3）规范RNA结合蛋白（RBP）与MAV相互作用。该建议旨在 定义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，我将通过通过 生化，基于测序和定量蛋白质组学方法以及病毒学技术 需要研究RNA病毒和先天免疫。在AIM 1中，我将定义RNA依赖性相互作用 在K99阶段的MAVS信号体组件之间 在R00相的RLR信号传导过程中使用RNA。通过AIM 2，我将确定所需的MAV中的序列 为了进行RNA的联系，并在K99阶段确定了一组MAVS结合的RNA。在 R00阶段，我将在MAVS信号体处阐述这些RNA的作用。在AIM 3中，我将筛选 与MAVS相关的RBP的入围名单，以识别那些在K99阶段影响MAV信号的RBP，并且 在抗病毒信号传导中确定三个RBP（Stau1，hnrnpl和RBM10）的分子功能 R00阶段。这些实验的总体结果将是定义以RNA为中心的新原则 MAVS信号体是组织的。了解RNA分子如何影响抗病毒信号可以解锁 针对病毒疾病以及自身免疫性疾病的新宿主指导的治疗策略。此外 我的顾问Savan博士，我在我的不同方面组成了一个具有专业知识的咨询委员会 研究。华盛顿大学的这种出色的培训环境将增加我的研究 在指导阶段，并为我提供过渡到独立学术所需的技能 研究先天免疫过程的RNA调节的研究人员。