title: Parvovirus Capsid Structures Required for Infection: Mutations Controlling Receptor Recognition and Protease Cleavages.
abstract: Parvovirus capsids are small but complex molecular machines responsible for undertaking many of the steps of cell infection, genome packing, and cell-to-cell as well as host-to-host transfer. The details of parvovirus infection of cells are still not fully understood, but the processes must involve small changes in the capsid structure that allow the endocytosed virus to escape from the endosome, pass through the cell cytoplasm, and deliver the single-stranded DNA (ssDNA) genome to the nucleus, where viral replication occurs. Here, we examine capsid substitutions that eliminate canine parvovirus (CPV) infectivity and identify how those mutations changed the capsid structure or altered interactions with the infectious pathway. Amino acid substitutions on the exterior surface of the capsid (Gly299Lys/Ala300Lys) altered the binding of the capsid to transferrin receptor type 1 (TfR), particularly during virus dissociation from the receptor, but still allowed efficient entry into both feline and canine cells without successful infection. These substitutions likely control specific capsid structural changes resulting from TfR binding required for infection. A second set of changes on the interior surface of the capsid reduced viral infectivity by >100-fold and included two cysteine residues and neighboring residues. One of these substitutions, Cys270Ser, modulates a VP2 cleavage event found in ∼10% of the capsid proteins that also was shown to alter capsid stability. A neighboring substitution, Pro272Lys, significantly reduced capsid assembly, while a Cys273Ser change appeared to alter capsid transport from the nucleus. These mutants reveal additional structural details that explain cell infection processes of parvovirus capsids.
  
  
  OBJECTIVE
  Parvoviruses are commonly found in both vertebrate and invertebrate animals and cause widespread disease. They are also being developed as oncolytic therapeutics and as gene therapy vectors. Most functions involved in infection or transduction are mediated by the viral capsid, but the structure-function correlates of the capsids and their constituent proteins are still incompletely understood, especially in relation to identifying capsid processes responsible for infection and release from the cell. Here, we characterize the functional effects of capsid protein mutations that result in the loss of virus infectivity, giving a better understanding of the portions of the capsid that mediate essential steps in successful infection pathways and how they contribute to viral infectivity.
authors.affiliation: Baker Institute for Animal Health, Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York, USA.
authors.forename: Heather M
authors.initials: HM
authors.lastname: Callaway
authors.affiliation: Baker Institute for Animal Health, Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York, USA.
authors.forename: Kurtis H
authors.initials: KH
authors.lastname: Feng
authors.affiliation: School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, New York, USA.
authors.forename: Donald W
authors.initials: DW
authors.lastname: Lee
authors.affiliation: Baker Institute for Animal Health, Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York, USA.|Department of Biomedical Sciences and Pathobiology, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, USA.
authors.forename: Andrew B
authors.identifier: http://orcid.org/0000-0003-0971-1215
authors.initials: AB
authors.lastname: Allison
authors.affiliation: Department of Biochemistry and Molecular Biology, Center for Structural Biology, McKnight Brain Institute, University of Florida, Gainesville, Florida, USA.
authors.forename: Melissa
authors.initials: M
authors.lastname: Pinard
authors.affiliation: Department of Biochemistry and Molecular Biology, Center for Structural Biology, McKnight Brain Institute, University of Florida, Gainesville, Florida, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: McKenna
authors.affiliation: Department of Biochemistry and Molecular Biology, Center for Structural Biology, McKnight Brain Institute, University of Florida, Gainesville, Florida, USA.
authors.forename: Mavis
authors.initials: M
authors.lastname: Agbandje-McKenna
authors.affiliation: Department of Medicine, Pennsylvania State University College of Medicine, Hershey, Pennsylvania, USA.
authors.forename: Susan
authors.initials: S
authors.lastname: Hafenstein
authors.affiliation: Baker Institute for Animal Health, Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York, USA crp3@cornell.edu.
authors.forename: Colin R
authors.identifier: http://orcid.org/0000-0002-1836-6655
authors.initials: CR
authors.lastname: Parrish
mesh_terms: D000595:Amino Acid Sequence; D036022:Capsid Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010450:Endopeptidases / Q000378:metabolism; D054884:Host-Pathogen Interactions; D008958:Models, Molecular; D009154:Mutation; D010322:Parvoviridae Infections / Q000821:virology*; D017992:Parvovirus / Q000502:physiology*; D011487:Protein Conformation; D021381:Protein Transport; D059748:Proteolysis; D011991:Receptors, Virus / Q000378:metabolism; D013329:Structure-Activity Relationship; D053585:Virus Attachment
chemical_list: D036022:Capsid Proteins; D011991:Receptors, Virus; D010450:Endopeptidases
keywords: parvovirus; proteolytic cleavage; receptor binding; structural virology; virus-receptor interaction