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Functions of the reovirus capsid

呼肠孤病毒衣壳的功能

基本信息

批准号：
10091191
负责人：
Pranav Danthi
金额：
$ 6.33万
依托单位：
TRUSTEES OF INDIANA UNIVERSITY
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：
2014-08-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10091191
关键词：
Affect Affinity Chromatography Antibodies Biochemical Biochemical Genetics Biological Assay Biology Capsid Capsid Proteins Cell Culture Techniques Cells Cryoelectron Microscopy Cytoplasm Data Disease Double-Stranded RNA Evolution Frequencies Generations Genetic Materials Genetic study Immune system Individual Infection Lipids Maintenance Mammalian Orthoreovirus Mass Spectrum Analysis Measures Mediating Membrane Membrane Lipids Molecular Molecular Conformation Mutagenesis Myocarditis Nucleosome Core Particle Penetration Peptide Hydrolases Peptides Physiological Positioning Attribute Process Property Proteins Proteolysis Publishing Reassortant Viruses Recovery Reovirus Research Role Shapes Structure System Time Variant Viral Viral Genome Viral Pathogenesis Viral Physiology Virion Virus Virus Assembly Virus Diseases Virus Replication Work base co-infection conformational conversion genetic analysis in vivo insight interdisciplinary approach mammalian genome microbiota mouse model multidisciplinary mutant nanodisk particle particle exposure recruit stoichiometry success tool viral genomics viral transmission

项目摘要

The proposed research uses mammalian reovirus (MRV) to define how a single capsid protein, μ1, influences distinct functions of the viral capsid. MRV particles are comprised of two concentric protein shells, the outer capsid and the inner core. The μ1 protein is a major component of the outer capsid. Published evidence and our preliminary data indicate that μ1 performs at least three functions. First, even though the two proteins are not in physical contact, properties of μ1 influence virus assembly in a way that alters the presentation of the sigma1 attachment protein on the virion. Second, two μ1 peptides, generated during virus disassembly, cooperate with host lipid membranes to facilitate further uncoating of the virus and permeabilize host membranes. Third, μ1 influences inter-particle interactions to form multivirion infectious units. With the aid of the following three Aims, the proposed research seeks to provide insight into the functions of μ1. In Aim 1, the role of μ1 in maintaining assembly fidelity will be determined. The position of sigma1 on the particle will be determined by biochemical studies and cryoelectron microscopy. The contribution of altering the strength of interaction between μ1 and adjacent capsid proteins on the presentation of sigma1 will be evaluated using genetic analysis. How changes to sigma1 encapsidation and sigma1 conformation influence virus replication in vivo will be determined using a mouse model of viral disease. In Aim 2, the function of μ1 in delivering core particles into the host cytoplasm will be defined. How μ1 peptides recruit entry intermediates to membranes will be determined by biochemical and genetic studies. The minimal number of μ1 peptides needed for successful recruitment of a virus entry intermediate, for pore formation and for successful infection will be quantified. The structure of the virus entry intermediate associated with the membrane will be determined by cryoelectron microscopy. Host proteins that associate with capsids following disassembly and influence the efficiency of infection will be identified by affinity purification and mass spectrometry. In Aim 3, the contribution of μ1 to MVIU formation will be identified. Regions important for inter-particle interactions will be determined by limited proteolysis and mass spectrometry. The relationship between MVIU formation, coinfection efficiency, and reassortment frequency will be determined. Whether MVIU formation also determines reassortment in vivo and if the determinants of MVIU formation and those that influence the recovery of reassortant progeny correlate, will be determined. Completion of this work will provide comprehensive insight into how μ1 completes each of these functions and define the properties of μ1 that influence the capacity of MRV to replicate in cell culture, produce reassortant progeny, and elicit disease.

这项拟议的研究使用哺乳动物呼肠孤病毒（MRV）来定义单个衣壳蛋白μ1如何