title: Capsid protein VP4 of human rhinovirus induces membrane permeability by the formation of a size-selective multimeric pore.
abstract: Non-enveloped viruses must deliver their viral genome across a cell membrane without the advantage of membrane fusion. The mechanisms used to achieve this remain poorly understood. Human rhinovirus, a frequent cause of the common cold, is a non-enveloped virus of the picornavirus family, which includes other significant pathogens such as poliovirus and foot-and-mouth disease virus. During picornavirus cell entry, the small myristoylated capsid protein VP4 is released from the virus, interacts with the cell membrane and is implicated in the delivery of the viral RNA genome into the cytoplasm to initiate replication. In this study, we have produced recombinant C-terminal histidine-tagged human rhinovirus VP4 and shown it can induce membrane permeability in liposome model membranes. Dextran size-exclusion studies, chemical crosslinking and electron microscopy demonstrated that VP4 forms a multimeric membrane pore, with a channel size consistent with transfer of the single-stranded RNA genome. The membrane permeability induced by recombinant VP4 was influenced by pH and was comparable to permeability induced by infectious virions. These findings present a molecular mechanism for the involvement of VP4 in cell entry and provide a model system which will facilitate exploration of VP4 as a novel antiviral target for the picornavirus family.
authors.affiliation: The Pirbright Institute, Pirbright, Surrey, United Kingdom; School of Molecular and Cellular Biology & Astbury Centre for Structural Molecular Biology, Faculty of Biological Sciences, University of Leeds, West Yorkshire, United Kingdom.
authors.forename: Anusha
authors.initials: A
authors.lastname: Panjwani
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, United States of America.
authors.forename: Mike
authors.initials: M
authors.lastname: Strauss
authors.affiliation: The Pirbright Institute, Pirbright, Surrey, United Kingdom.
authors.forename: Sarah
authors.initials: S
authors.lastname: Gold
authors.affiliation: The Pirbright Institute, Pirbright, Surrey, United Kingdom.
authors.forename: Hannah
authors.initials: H
authors.lastname: Wenham
authors.affiliation: The Pirbright Institute, Pirbright, Surrey, United Kingdom.
authors.forename: Terry
authors.initials: T
authors.lastname: Jackson
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, United States of America.
authors.forename: James J
authors.initials: JJ
authors.lastname: Chou
authors.affiliation: School of Molecular and Cellular Biology & Astbury Centre for Structural Molecular Biology, Faculty of Biological Sciences, University of Leeds, West Yorkshire, United Kingdom.
authors.forename: David J
authors.initials: DJ
authors.lastname: Rowlands
authors.affiliation: School of Molecular and Cellular Biology & Astbury Centre for Structural Molecular Biology, Faculty of Biological Sciences, University of Leeds, West Yorkshire, United Kingdom.
authors.forename: Nicola J
authors.initials: NJ
authors.lastname: Stonehouse
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, United States of America.
authors.forename: James M
authors.initials: JM
authors.lastname: Hogle
authors.affiliation: The Pirbright Institute, Pirbright, Surrey, United Kingdom.
authors.forename: Tobias J
authors.initials: TJ
authors.lastname: Tuthill
mesh_terms: D015153:Blotting, Western; D036022:Capsid Proteins / Q000378:metabolism*; D002463:Cell Membrane Permeability; D006367:HeLa Cells; D006801:Humans; D008081:Liposomes; D046529:Microscopy, Electron, Transmission; D011994:Recombinant Proteins / Q000378:metabolism; D012229:Rhinovirus / Q000378:metabolism / Q000472:pathogenicity*
chemical_list: D036022:Capsid Proteins; D008081:Liposomes; D011994:Recombinant Proteins
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