title: Matrix proteins of Nipah and Hendra viruses interact with beta subunits of AP-3 complexes.
abstract: Paramyxoviruses and other negative-strand RNA viruses encode matrix proteins that coordinate the virus assembly process. The matrix proteins link the viral glycoproteins and the viral ribonucleoproteins at virus assembly sites and often recruit host machinery that facilitates the budding process. Using a co-affinity purification strategy, we have identified the beta subunit of the AP-3 adapter protein complex, AP3B1, as a binding partner for the M proteins of the zoonotic paramyxoviruses Nipah virus and Hendra virus. Binding function was localized to the serine-rich and acidic Hinge domain of AP3B1, and a 29-amino-acid Hinge-derived polypeptide was sufficient for M protein binding in coimmunoprecipitation assays. Virus-like particle (VLP) production assays were used to assess the relationship between AP3B1 binding and M protein function. We found that for both Nipah virus and Hendra virus, M protein expression in the absence of any other viral proteins led to the efficient production of VLPs in transfected cells, and this VLP production was potently inhibited upon overexpression of short M-binding polypeptides derived from the Hinge region of AP3B1. Both human and bat (Pteropus alecto) AP3B1-derived polypeptides were highly effective at inhibiting the production of VLPs. VLP production was also impaired through small interfering RNA (siRNA)-mediated depletion of AP3B1 from cells. These findings suggest that AP-3-directed trafficking processes are important for henipavirus particle production and identify a new host protein-virus protein binding interface that could become a useful target in future efforts to develop small molecule inhibitors to combat paramyxoviral infections.
  
  
  OBJECTIVE
  Henipaviruses cause deadly infections in humans, with a mortality rate of about 40%. Hendra virus outbreaks in Australia, all involving horses and some involving transmission to humans, have been a continuing problem. Nipah virus caused a large outbreak in Malaysia in 1998, killing 109 people, and smaller outbreaks have since occurred in Bangladesh and India. In this study, we have defined, for the first time, host factors that interact with henipavirus M proteins and contribute to viral particle assembly. We have also defined a new host protein-viral protein binding interface that can potentially be targeted for the inhibition of paramyxovirus infections.
authors.affiliation: Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, Pennsylvania, USA.
authors.forename: Weina
authors.initials: W
authors.lastname: Sun
authors.affiliation: Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, Pennsylvania, USA.
authors.forename: Thomas S
authors.initials: TS
authors.lastname: McCrory
authors.affiliation: Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, Pennsylvania, USA.
authors.forename: Wei Young
authors.initials: WY
authors.lastname: Khaw
authors.affiliation: Department of Microbiology and Immunology, Uniformed Services University, Bethesda, Maryland, USA.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Petzing
authors.affiliation: Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, Pennsylvania, USA.
authors.forename: Terrell
authors.initials: T
authors.lastname: Myers
authors.affiliation: Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, Pennsylvania, USA Center for Molecular Immunology and Infectious Disease, The Pennsylvania State University, University Park, Pennsylvania, USA aps13@psu.edu.
authors.forename: Anthony P
authors.initials: AP
authors.lastname: Schmitt
mesh_terms: D033963:Adaptor Protein Complex 3 / Q000378:metabolism*; D033966:Adaptor Protein Complex beta Subunits / Q000378:metabolism*; D045404:Hendra Virus / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D045405:Nipah Virus / Q000502:physiology*; D025941:Protein Interaction Mapping; D014763:Viral Matrix Proteins / Q000378:metabolism*; D057074:Virus Release
chemical_list: C439973:AP3B1 protein, human; D033963:Adaptor Protein Complex 3; D033966:Adaptor Protein Complex beta Subunits; D014763:Viral Matrix Proteins