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）：副粘病毒是通过直接与细胞膜融合进入细胞的有包膜病毒。在进入过程中，两种病毒表面糖蛋白 HN（受体结合分子）和 F（病毒融合蛋白）以高度特异性的方式合作，介导受体结合后的融合。然而，在病毒复制和在细胞质中组装的过程中，融合过程是沉默的，并且在传入的病毒遇到其目标细胞之前，融合机器是不活动的。融合过程在时间和地点上如何协调？为了了解这些机制并阐明副粘病毒如何进入细胞，我们研究了人类副流感病毒（HPIV），它是儿童呼吸道疾病的重要原因。我们的结果揭示了受体结合蛋白在副粘病毒融合中的基本作用，以及病毒生命周期中糖蛋白之间协调相互作用的原理。我们开发了新的跨学科工具来剖析副粘病毒控制进入的复杂过程，现在可以解决该领域的核心问题。具体目标 1. 副粘病毒受体结合分子的多种功能协调的机制。 HN 的主要受体结合位点的附着如何转化为 HN 的第二结合/触发位点促进的 F 触发？ HN 两个功能结合位点的突变将用于研究 HN 如何触发 F。这些实验将确定 HN 的活性如何针对 HPIV 进行协调，以及其他副粘病毒中的不同策略如何实现相同的目的。具体目标2.受体结合分子和融合分子在融合和进入过程中如何通讯。在融合和进入过程中，HN 和 F 之间的通信本质是什么？这种通信是如何调节的？一系列创新策略将检验特定 HN-F 相互作用调节 F 激活的假设。这些实验将首次定义构成副粘病毒融合机的分子之间的生物学相关对话。具体目标 3. 气道细胞中副粘病毒生命周期中融合的调节。 F-激活的正确时机对于进入至关重要；对于感染，只有当 F 与靶细胞膜接触时才会发生触发。这些蛋白质的不同功能在病毒生命周期中是如何受到调节的，我们是否可以颠覆这种调节作为抗病毒药物的策略？我们将在代表人类肺上皮的系统中测试 F 触发失调会阻止成功感染的假设。我们将确定化合物是否可以过早触发 F，以及这是否可以预防气道感染，从而提供新的抗病毒策略。这里揭示的基本原理将为需要变构激活的复杂分子机器提供见解。结果可能适用于其他病原体，这些病原体必须仅在特定时间和地点触发这些关键的进入活动以引发感染。公共卫生相关性：这项研究提案将有助于更好地了解副粘病毒如何进入人体细胞引发感染。新发现的参与进入过程的机制可以作为抗病毒药物治疗或预防人类呼吸道疾病的潜在新靶标。