title: C-Terminal DxD-Containing Sequences within Paramyxovirus Nucleocapsid Proteins Determine Matrix Protein Compatibility and Can Direct Foreign Proteins into Budding Particles.
abstract: Paramyxovirus particles are formed by a budding process coordinated by viral matrix (M) proteins. M proteins coalesce at sites underlying infected cell membranes and induce other viral components, including viral glycoproteins and viral ribonucleoprotein complexes (vRNPs), to assemble at these locations from which particles bud. M proteins interact with the nucleocapsid (NP or N) components of vRNPs, and these interactions enable production of infectious, genome-containing virions. For the paramyxoviruses parainfluenza virus 5 (PIV5) and mumps virus, M-NP interaction also contributes to efficient production of virus-like particles (VLPs) in transfected cells. A DLD sequence near the C-terminal end of PIV5 NP protein was previously found to be necessary for M-NP interaction and efficient VLP production. Here, we demonstrate that 15-residue-long, DLD-containing sequences derived from either the PIV5 or Nipah virus nucleocapsid protein C-terminal ends are sufficient to direct packaging of a foreign protein, Renilla luciferase, into budding VLPs. Mumps virus NP protein harbors DWD in place of the DLD sequence found in PIV5 NP protein, and consequently, PIV5 NP protein is incompatible with mumps virus M protein. A single amino acid change converting DLD to DWD within PIV5 NP protein induced compatibility between these proteins and allowed efficient production of mumps VLPs. Our data suggest a model in which paramyxoviruses share an overall common strategy for directing M-NP interactions but with important variations contained within DLD-like sequences that play key roles in defining M/NP protein compatibilities.
  
  
  OBJECTIVE
  Paramyxoviruses are responsible for a wide range of diseases that affect both humans and animals. Paramyxovirus pathogens include measles virus, mumps virus, human respiratory syncytial virus, and the zoonotic paramyxoviruses Nipah virus and Hendra virus. Infectivity of paramyxovirus particles depends on matrix-nucleocapsid protein interactions which enable efficient packaging of encapsidated viral RNA genomes into budding virions. In this study, we have defined regions near the C-terminal ends of paramyxovirus nucleocapsid proteins that are important for matrix protein interaction and that are sufficient to direct a foreign protein into budding particles. These results advance our basic understanding of paramyxovirus genome packaging interactions and also have implications for the potential use of virus-like particles as protein delivery tools.
authors.affiliation: Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, Pennsylvania, USA.
authors.forename: Greeshma
authors.initials: G
authors.lastname: Ray
authors.affiliation: Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, Pennsylvania, USA.
authors.forename: Phuong Tieu
authors.initials: PT
authors.lastname: Schmitt
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: D020816:Amino Acid Motifs; D002460:Cell Line; D006801:Humans; D049410:Luciferases, Renilla / Q000378:metabolism; D009109:Mumps virus / Q000235:genetics / Q000502:physiology*; D045405:Nipah Virus / Q000235:genetics / Q000502:physiology*; D019590:Nucleocapsid Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D045402:Parainfluenza Virus 5 / Q000235:genetics / Q000502:physiology*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000378:metabolism*; D022701:Virosomes / Q000378:metabolism; D019065:Virus Assembly; D057074:Virus Release
chemical_list: D019590:Nucleocapsid Proteins; D014763:Viral Matrix Proteins; D022701:Virosomes; D049410:Luciferases, Renilla