Strain Dependent Structure and Function of the Influenza NS1 Protein

流感 NS1 蛋白的菌株依赖性结构和功能

• 批准号: 10053291
• 负责人: Chad Petit
• 金额: $ 37.13万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-11-16 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10053291
• 关键词: Affect; Allosteric Regulation; Amino Acids; Animals; Antiviral Agents; Antiviral Response; Automobile Driving; Binding; Cells; Cleavage And Polyadenylation Specificity Factor; Crystallization; Data; Development; Disease; Drug Design; Drug Targeting; Epidemic; Future; Genes; Genetic Structures; Goals; Health; Human; Immune; Immune response; Impairment; Infection; Influenza; Innate Immune Response; Interferon-beta; Interferons; Investigation; Knowledge; Measures; Mission; Molecular; Molecular Structure; Morbidity - disease rate; Motion; NMR Spectroscopy; Nonstructural Protein; Pathogenicity; Play; Production; Property; Protein Dynamics; Proteins; Public Health; RNA Recognition Motif; Recombinants; Relaxation; Research; Resolution; Role; Sampling; Signal Transduction; Structural Models; Structure; Structure-Activity Relationship; Testing; Tretinoin; United States National Institutes of Health; Variant; Viral; Virulence; Virus Replication; anti-influenza; base; experimental study; influenza infection; influenza virulence; influenza virus strain; influenzavirus; inhibitor/antagonist; innovation; millisecond; mortality; mutant; novel; nucleocytoplasmic transport; pandemic disease; pathogen; resistant strain; response; small molecule

Developing effective antiviral drugs requires a detailed understanding of the molecular mechanisms underlying the targeted host-pathogen interaction. Specifically, precise structural models of these interactions can provide mechanistic details with atomic resolution to assist in the efficient development of novel compounds against pathogens of significant health relevance. The influenza virus is a prime example of one such pathogen. Novel strains of the influenza virus develop annually via infection and replication in a number of animal hosts, including humans. The relative ability of these strains to cause disease, or virulence, is determined by a number of interactions between viral and cellular proteins. Although influenza non-structural protein 1 (NS1) is known to play a critical role in virulence, there is a fundamental gap in our knowledge of the genetic and structural determinants that facilitate the multiple strain-dependent functions attributed to NS1 in the host cell. It is therefore our long-term goal to understand the molecular mechanisms that underlie strain-dependent function of NS1. The objective of this application is to structurally characterize interactions with two cellular proteins (CPSF30 and RIG-I) that are important for the activation of the innate immune response. Our central hypothesis is that structural and dynamic features unique to certain NS1 variants account for the diverse array of functions attributed to NS1. The rationale that underlies the proposed research is that elucidating structure- function relationships between NS1 and its cellular interaction partners will aid in the development of antiviral drugs that target these critical interactions known to modulate virulence. Our central hypothesis will be tested by pursuing three specific aims: 1) structurally and functionally characterize the multiple interactions between NS1 and RIG-I, 2) determine the role of microsecond-millisecond (µs-ms) motions in proper function of the NS1 effector domain (NS1ED), and 3) determine the mechanism of action by which JJ3297 suppresses influenza replication. In Aim 1, NMR spectroscopy and mutant recombinant influenza viruses will be used to structurally and functionally characterize the multiple interactions between NS1 and RIG-I. In Aim 2, relaxation dispersion experiments will be used to determine the role that protein dynamics play in facilitating the interaction between the NS1ED and CPSF30 and intracellular localization of NS1. In Aim 3, NMR spectroscopy will be used to determine the mechanism of action by which JJ3297 suppresses influenza replication. Our innovative approach will be the first investigation into how protein dynamics and strain specific structural variations facilitate proper function of NS1 in the context of viral replication and pathogenicity. This will also be the first systematic study to determine functional variations in NS1 between multiple strains of influenza. The proposed research is significant because it will define the molecular mechanisms underlying NS1 functions shown to modulate influenza virulence. By defining these molecular mechanisms, this proposal will inform efforts in developing influenza antiviral drugs targeting NS1, thereby supporting the overall mission of the NIH.

开发有效的抗病毒药物需要详细了解目标宿主 - 病原体相互作用的分子机制。具体而言，这些相互作用的精确结构模型可以通过原子分辨率提供机械细节，以帮助有效地开发针对重要健康相关病原体的新型化合物。影响者病毒是这种病原体的一个主要例子。新颖的影响者病毒每年通过感染和复制在包括人类在内的许多动物宿主中复制。这些菌株引起疾病或病毒的相对能力取决于病毒和细胞蛋白之间的多种相互作用。尽管众所周知，有影响力的非结构蛋白1（NS1）在病毒中起关键作用，但在我们对遗传和结构决定剂的了解中存在一个基本差距，可以促进促进宿主细胞中NS1的多重菌株依赖性功能。因此，我们的长期目标是了解NS1应变依赖性功能的分子机制。该应用的目的是在结构上表征与两个细胞蛋白（CPSF30和RIG-I）相互作用，这对于激活先天免疫响应很重要。我们的中心假设是，某些NS1变体所独有的结构和动态特征解释了归因于NS1的潜水员阵列。拟议的研究基础的基本原理是阐明NS1与其细胞相互作用伙伴之间的结构 - 功能关系将有助于开发抗病毒药物，这些抗病毒药是针对这些已知可调节病毒的这些关键相互作用的抗病毒药。我们的中心假设将通过追求三个具体目的来检验：1）在结构和功能上表征NS1和RIG-I之间的多个相互作用，2）确定微秒毫秒（µS-MS）运动在NS1效应域（NS1ED）（NS1ED）的正常功能中的作用（µS-MS）运动，以及3）抑制了JJ3297的作用机制。在AIM 1中，NMR光谱和突变重组影响病毒将在结构和功能上表征NS1和RIG-I之间的多个相互作用。在AIM 2中，将使用松弛分散实验来确定蛋白质动力学在促进NS1ED与CPSF30之间的相互作用以及NS1的细胞内定位中起作用的作用。在AIM 3中，NMR光谱法将用于确定JJ3297抑制影响力复制的作用机理。我们的创新方法将是对蛋白质动力学和应变特定结构变化如何在病毒复制和致病性背景下促进NS1的适当功能的首次投资。这也将是确定多种影响力菌株之间NS1功能变化的首次系统研究。拟议的研究很重要，因为它将定义用于调节影响力病毒的NS1功能的分子机制。通过定义这些分子机制，该提案将为开发针对NS1的抗病毒药物而努力，从而支持NIH的整体任务。