Structural and mechanistic studies of self and non-self recognition by RIG-I

RIG-I对自我和非自我识别的结构和机制研究

基本信息

批准号：
9282445
负责人：
Gary A. Brewer
金额：
$ 42.25万
依托单位：
RUTGERS, THE STATE UNIV OF N.J.
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-15 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9282445
关键词：
ATP Hydrolysis ATP phosphohydrolase Address Affect Affinity Anti-Inflammatory Agents Anti-inflammatory Antiviral Agents Area Autoimmune Process Binding Biochemical Biological Assay Biophysics Caspase Cells Collection Complement Complex Crystallization Cytoplasm Cytoplasmic Receptors Dengue Deuterium Development Disease Disulfides Double-Stranded RNA Ducks Ebola virus Enzymatic Biochemistry Failure Fluorescence Gene Silencing Genes Goals Grant Hepatitis C Human Hydrogen Immune Evasion Immune response Immune signaling Infection Influenza Innate Immune Response Innate Immune System Kinetics Laboratories Lead Length Measures Methods Modeling Modification Molecular Molecular Conformation Monitor Movement Natural Immunity Outcome Pathogenicity Pathway interactions Pattern Pattern Recognition Physiological Play Post-Translational Modification Site Process Property Proteins Publishing RNA RNA Binding RNA Virus Infections Reaction Reagent Recruitment Activity Reoviridae Reovirus Research Project Grants Resolution Role Series Signal Transduction Solvents Structure Systemic infection Testing Therapeutic Thermodynamics Tretinoin Viral Virus Virus Diseases West Nile virus X-Ray Crystallography analog base design helicase insight microbial mutant pathogen prevent public health relevance receptor research study respiratory structural biology therapeutic development tripolyphosphate viral RNA

项目摘要

DESCRIPTION (provided by applicant): The innate immune system is the first line of defense against microbial and viral infections. A failure to elicit the early innate immune response leads to systemic infections. A significant number of human viruses, including Influenza, Hepatitis C, Dengue, West Nile, Respiratory Syncytial, Reovirus, and Ebola are recognized by the innate immunity receptor RIG-I. The overall goal of this proposal is to understand how RIG-I (Retinoic Acid-inducible Gene-I), a cytoplasmic receptor discriminates normal, cellular from viral RNAs to stimulate a host response. A major goal of our collaborative research studies has been to understand the thermodynamic, kinetic, and structural mechanisms by which RIG-I recognizes atypical pathogen associated molecular pattern (PAMP) feature in RNAs. Our crystal structure of RIG-I bound to blunt-ended dsRNA and an ATP analog established a new paradigm for RIG-I activation. The goal of this proposal is to rigorously test the RIG-I activation model using carefully designed biochemical, kinetic, and structural studies. Our preliminary results demonstrate that RIG-I is regulated in multiple ways and RNA binding affinity is not the only criterion for PAMP selection. With a unique collection of purified protein, RNA reagents, we will employ complementary biochemical, biophysical, structural, and cell based approaches to 1) understand the basis of PAMP versus non-PAMP recognition of RNA by RIG-I; 2) characterize RIG-I ATPase activity and its role in RIG-I activation; and 3) understand the mechanism of RIG-I signaling. The outcomes are better understanding of self versus non-self recognition and RIG-I evasion mechanism, which can lead to the development of broad-spectrum antivirals, anti-inflammatory therapeutics and RNA-based gene silencing agents.

描述（由申请人提供）：先天免疫系统是针对微生物和病毒感染的第一道防线。未能引起先天免疫反应会导致系统性感染。先天免疫受体rig-I认可了许多人类病毒，包括流感，丙型肝炎，登革热，西尼罗河，呼吸道合胞病毒，依泊病毒和埃博拉病毒。该提案的总体目标是了解RIG-I（视黄酸诱导的基因I）是如何区分正常的，细胞受体与病毒RNA区分正常以刺激宿主反应。我们的协作研究的一个主要目标是了解RIG-I识别RNA中非典型致病原分（PAMP）特征的热力学，动力学和结构机制。我们的RIG-I的晶体结构与钝性的DSRNA和ATP模拟结合，建立了一种用于RIG-I激活的新范式。该建议的目的是使用精心设计的生化，动力学和结构研究严格测试RIG-I激活模型。我们的初步结果表明，RIG-1是通过多种方式调节的，RNA结合亲和力并不是PAMP选择的唯一标准。通过独特的纯化蛋白质RNA试剂的集合，我们将采用互补的生化，生物物理，结构和基于细胞的方法的方法，以了解aig-i对pAMP对RNA的识别的基础。 2）表征RIG-I ATPase活性及其在RIG-I激活中的作用； 3）了解RIG-I信号传导的机制。结果是对自我与非自我识别和RIG-I逃避机制的更好理解，这可能导致广谱抗病毒药，抗炎疗法和基于RNA的基因沉默剂的发展。