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

项目摘要/摘要 控制好抗病毒先天免疫是限制病毒病原体和防止异常的关键 发炎。一个关键的抗病毒信号通路是由RIG-I样受体(RLRs)启动的，RLRs可以感知病毒RNA 在胞浆中通过以细胞外信号转导系统为中心的高级信号平台诱导干扰素的产生 接头蛋白MAVs。虽然MAV信号小体的组成成分是碱基表达的，但RLR 只有当它们在内质网线粒体接触部位聚集在小牛周围时，信号才会继续。许多人 小牛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。然而，蛋白质复合体中的功能相互作用可以由RNA分子介导， RNA分子是否通过MAV信号小体在信号转导中发挥作用尚不清楚。我有过 发现1)MAV与非病毒RNA有关，2)RNA通过MAV促进信号转导 信号小体；3)典型的RNA结合蛋白(RBPs)与MAV相互作用。这项建议旨在 明确RNA和限制性商业惯例在通过MAVS信号体调节抗病毒信号中的功能作用。 在指导阶段，我将通过以下方式获得关于RNA-蛋白质相互作用特征的新培训 生化、基于测序和定量的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。在目标1中，我将定义依赖于RNA的相互作用 在K99期的MAVS信号小体的组件之间，并确定与 在R00阶段RLR信号转导过程中与RNA结合。通过目标2，我将精确定位小牛所需的序列 对于RNA结合，并在K99阶段确定一组高置信度的MAV结合的RNA。在.期间 R00阶段，我将从功能上剖析这些RNA在小牛信号体中的作用。在《目标3》中，我会放映 与MAV相关的RBP的入围名单，以确定在K99阶段影响MAV信号的那些RBP，以及 确定三种限制性商业惯例(STAU1、HNRNPL和RBM10)在抗病毒信号转导中的分子功能 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，通过这些原则 小牛信号体是有组织的。了解RNA分子如何影响抗病毒信号传递可以解开 针对病毒疾病和自身免疫性疾病的新的宿主导向治疗策略。除了……之外 我的顾问萨万博士，我已经组建了一个咨询委员会，该委员会在我的 研究。总之，华盛顿大学的这种良好的培训环境将加强我的研究 在指导阶段，并让我具备过渡到独立学术人员所需的技能 研究先天免疫过程的RNA调节的研究人员。

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.