Paramyxovirus F protein mediated membrane fusion

副粘病毒F蛋白介导的膜融合

基本信息

批准号：
8137427
负责人：
Rebecca E. Dutch
金额：
$ 4.61万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-13 至 2012-09-12
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8137427
关键词：
Address Affect Alanine Antiviral Agents Antiviral Therapy Cathepsin L Cell fusion Cell membrane Cells Charge Chimeric Proteins Cleaved cell Cysteine Cytoplasmic Tail Data Development Electrostatics Endocytosis Event Family Family member Funding Glycine Glycoproteins Goals Hendra Virus Human Human Metapneumovirus Human respiratory syncytial virus Integral Membrane Protein Kinetics Lead Measles virus Mediating Membrane Fusion Molecular Mutation Nipah Virus Paramyxovirus Peptide Hydrolases Peptides Play Positioning Attribute Process Proline Proteins Reagent Recombinants Recycling Research Respiratory syncytial virus Role Route Scanning Staging Structure System Transmembrane Domain Variant Viral Viral Proteins Virus insight mutant novel parainfluenza virus pathogen protein activation protein folding public health relevance recombinant virus trafficking

项目摘要

DESCRIPTION (provided by applicant): Membrane fusion, a process critical for viral entry, is promoted by the paramyxovirus fusion (F) proteins. All F proteins contain a number of common features that play fundamental roles in fusion. However, significant variations exist between F proteins related to protein activation and promotion of membrane fusion, and many critical questions remain concerning the mechanism by which fusion is promoted by these important viral proteins. The long-term objective of our research is to understand the precise mechanism(s) of paramyxovirus F protein-promoted membrane fusion. Our overall hypothesis is that the function of domains critical for fusion promotion will be conserved in diverse F proteins, but that interactions in these domains will modulate triggering mechanisms. To address this important hypothesis we will pursue the following specific aims: 1.) We will build on our exciting preliminary data which demonstrates trimer formation of isolated Hendra F transmembrane (TM) domains to define the role of TM-TM interactions in glycoprotein folding and fusion. We will therefore delineate the residues critical for TM-TM interactions, and evaluate the role of this interaction in Hendra F folding, trafficking and fusion; analyze the effect of HRB or cytoplasmic-tail additions on stability of TM-TM interactions; compare TM-TM interactions for the PIV5 and HMPV F proteins to those observed for the Hendra F TM; and determine if TM domains from paramyxovirus attachment proteins interact with themselves or with the F protein TM domains; 2.) We will define the role of low pH and endocytosis in HMPV viral entry and delineate the role of electrostatic repulsion in the HRB linker region in HMPV F low pH-induced conformational changes and entry of recombinant viruses and; 3.) We will analyze the role of stabilizing interactions between HRA and a conserved region of F2, which our studies of PIV5 F indicate can play a role in triggering, by defining key interactions in this domain in PIV5 F, assessing the stage of fusion which is affected, analyzing the effect of mutations in this region on viral entry, and analyzing the effect of mutations to this region in other F proteins. Accomplishing these goals will provide crucial information on the regions that control the molecular events involved in the F protein-promoted membrane fusion process, and potentially identify new targets for antiviral therapy. PUBLIC HEALTH RELEVANCE The paramyxovirus family contains both established human pathogens, such as measles virus and respiratory syncytial virus, and newly emerged human pathogens, including the highly pathogenic Hendra and Nipah viruses and the recently identified human metapneumovirus (HMPV). A detailed understanding of the fusion mechanism could lead to development of new antiviral reagents.

描述（由申请人提供）：膜融合是病毒输入至关重要的过程，由帕托粘病毒融合（F）蛋白促进。所有F蛋白都包含许多在融合中扮演基本角色的常见特征。然而，与蛋白质激活和促进膜融合有关的F蛋白之间存在显着变化，并且关于这些重要病毒蛋白促进融合的机制的许多关键问题仍然存在。我们研究的长期目标是了解帕托毒素F蛋白促进膜融合的精确机制。我们的总体假设是，在不同的F蛋白中，对融合促进至关重要的域的功能将是保守的，但是这些域中的相互作用将调节触发机制。为了解决这一重要假设，我们将追求以下特定目的：1。）我们将基于令人兴奋的初步数据，该数据表明孤立的Hendra F跨膜（TM）域的三聚体形成，以定义TM-TM相互作用在糖蛋白折叠和融合中的作用。因此，我们将描述对TM-TM相互作用至关重要的残基，并评估这种相互作用在Hendra F折叠，贩运和融合中的作用；分析HRB或细胞质尾添加对TM-TM相互作用稳定性的影响；将PIV5和HMPV F蛋白的TM-TM相互作用与对Hendra F TM观察到的相互作用进行比较；并确定来自帕托病毒附着蛋白的TM结构域是否与自身或F蛋白TM结构域相互作用； 2.）我们将定义低pH和内吞作用在HMPV病毒进入中的作用，并描述HMPV f低pH诱导的重组病毒构象变化和重组病毒的输入的HRB接头区域中静电排斥的作用； 3.）我们将分析HRA与F2的保守区域之间稳定相互作用的作用，我们对PIV5 F的研究表明可以通过在PIV5 F中定义该域中的关键相互作用来在触发中发挥作用，从而评估了受影响的融合阶段，从而分析了该区域对病毒式进入的效果，并分析了该区域的效应，并分析了该区域的其他效果。实现这些目标将提供有关控制F蛋白促进膜融合过程的分子事件区域的重要信息，并有可能确定抗病毒治疗的新靶标。公共卫生相关性帕托马维病毒家族既包含既定的人类病原体，例如麻疹病毒和呼吸道综合病毒，以及新出现的人类病原体，包括高度致病性的亨德拉和尼帕病毒，以及最近确定的人类替代病毒（HMPV）。对融合机制的详细理解可能导致新的抗病毒试剂的发展。