Molecular basis for paramyxovirus entry

副粘病毒进入的分子基础

• 批准号: 8299252
• 负责人: Anne Moscona
• 金额: $ 8.76万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2012-08-05
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8299252
• 关键词: Active Sites; Address; Antiviral Agents; Binding; Binding Proteins; Binding Sites; Biological Assay; Bronchiolitis; Cell fusion; Cell membrane; Cells; Child; Chimeric Proteins; Cleaved cell; Communication; Complement; Complementary DNA; Complex; Croup; Cytoplasm; Epithelium; Family; Fluorescence; Glycoproteins; Health; Human; Human Cloning; Infection; Lead; Life Cycle Stages; Lung; Lung diseases; Mediating; Membrane Glycoproteins; Modeling; Molecular; Molecular Machines; Molecular Virology; Mutation; Nature; Neuraminidase; Outcome; Parainfluenza; Paramyxovirus; Pathway interactions; Peptides; Process; Property; Readiness; Receptor Cell; Regulation; Research Proposals; Role; Series; Site; System; Testing; Time; Translating; Vaccines; Variant; Viral; Viral Fusion Proteins; Virus; Virus Assembly; Virus Replication; Work; base; improved; inhibitor/antagonist; innovation; insight; meetings; member; mutant; novel therapeutics; parainfluenza virus; pathogen; premature; prevent; protein function; receptor; receptor binding; research study; small molecule; tool

DESCRIPTION (provided by applicant): Paramyxoviruses are enveloped viruses that enter cells by fusing directly with the cell membrane. During entry the two viral surface glycoproteins HN (the receptor-binding molecule) and F (the viral fusion protein) cooperate in a highly specific way to mediate fusion upon receptor binding. However, during virus replication and assembly in the cytoplasm the fusion process is silent, and until an incoming virus meets its target cell, the fusion machine is inactive. How is the fusion process coordinated in time and place? To understand these mechanisms and elucidate how paramyxoviruses enter cells, we study the human parainfluenza virus (HPIV), an important cause of respiratory disease in children. Our results have uncovered fundamental roles of the receptor binding protein in paramyxovirus fusion, and principles of coordinated interaction between the glycoproteins during the viral life cycle. We have developed new, interdisciplinary tools to dissect the complex process whereby paramyxoviruses control entry, and central questions in the field can now be addressed. Specific Aim 1. Mechanisms by which the diverse functions of the paramyxovirus receptor-binding molecule are coordinated. How does attachment by the primary receptor binding site of HN translate into the F-triggering that is promoted by HN's second binding/triggering site? Mutations at HN's two functional binding sites will be used to study how HN works to trigger F. These experiments will define how the activities of HN are coordinated for HPIV, and how distinct strategies in other paramyxoviruses accomplish the same ends. Specific Aim 2. How the receptor-binding molecule and the fusion molecule communicate during fusion and entry. What is the nature of the communication between HN and F during fusion and entry, and how is this communication regulated? A series of innovative strategies will test the hypothesis that specific HN-F interactions regulate F-activation. These experiments will for the first time define the biologically relevant dialog between the molecules that comprise the paramyxovirus fusion machine. Specific Aim 3. Regulation of fusion during the paramyxoviral life cycle in airway cells. Correct timing of F-activation is essential for entry; for infection, triggering must occur only when F is in contact with the target cell membrane. How are the diverse functions of these proteins regulated during the viral life cycle, and can we subvert this regulation as a strategy for antivirals? We will test the hypothesis that dysregulation of F-triggering precludes successful infection, in a system that represents human lung epithelium. We will determine whether compounds can prematurely trigger F, and whether this may prevent airway infection, providing a new antiviral strategy. The underlying principles revealed here will offer insights into complex molecular machines that require allosteric activation. The results will likely apply to other pathogens that must trigger these key entry activities only at a specific time and place to initiate infection. PUBLIC HEALTH RELEVANCE: This research proposal will lead to a better understanding of how paramyxoviruses enter human cells to initiate infection. Newly identified mechanisms involved in the entry process could serve as potential new targets for antivirals to treat or prevent human respiratory diseases.

描述（由申请人提供）：Paramyxovirus是包膜的病毒，通过直接与细胞膜融合来进入细胞。在进入期间，两个病毒表面糖蛋白HN（受体结合分子）和F（病毒融合蛋白）以高度特异性的方式进行了合作，以介导受体结合时融合融合。但是，在细胞质中的病毒复制和组装过程中，融合过程是静默的，直到传入的病毒符合其靶细胞，融合机是不活跃的。融合过程如何在时间和地点进行协调？为了理解这些机制并阐明了帕托病毒进入细胞的方式，我们研究了人类副菌病毒（HPIV），这是儿童呼吸道疾病的重要原因。我们的结果发现了受体结合蛋白在帕托维病毒融合中的基本作用，以及在病毒生命周期期间糖蛋白之间的协调相互作用的原理。我们已经开发了新的，跨学科的工具，以剖析复杂的过程，从而在该过程中进行了paramyxoviruses控制条目，现在可以解决现场中的中心问题。特定的目标1。丙糖病毒受体结合分子的各种功能是协调的机制。 HN的主要受体结合位点的附件如何转化为由HN的第二个结合/触发位点促进的F触发器？ HN的两个功能结合位点的突变将用于研究HN如何触发F。这些实验将定义HN的活性如何用于HPIV，以及其他帕托氏菌病毒的不同策略如何完成相同的目的。具体目标2。在融合和进入过程中，受体结合分子和融合分子如何通信。在融合和进入期间，HN和F之间的交流的性质是什么？这种通信如何受到监管？一系列创新策略将检验以下假设：特定的HN-F相互作用调节F-Acrovation。这些实验将首次定义构成帕托马病毒融合机的分子之间的生物学相关对话。具体目标3。气道细胞中氧化病毒生命周期期间融合的调节。正确的F-激活时间对于进入至关重要；对于感染，只有在F与靶细胞膜接触时，才必须发生触发。这些蛋白质在病毒生命周期期间如何调节这些蛋白质的各种功能，我们可以将这种调节作为抗病毒药的策略而颠覆？我们将检验以下假设：在代表人肺上皮的系统中，F触发的失调无法成功感染。我们将确定化合物是否可以过早触发F，以及这是否可以防止气道感染，从而提供新的抗病毒药策略。此处揭示的基本原理将提供对需要变构激活的复杂分子机器的见解。结果可能适用于仅在特定时间和地点启动感染的其他病原体。公共卫生相关性：这项研究建议将更好地理解帕托马病毒如何进入人类细胞以启动感染。进入过程中涉及的新确定的机制可以作为治疗或预防人类呼吸系统疾病的潜在新目标。