title: Ebola virus VP35-VP40 interaction is sufficient for packaging 3E-5E minigenome RNA into virus-like particles.
abstract: The packaging of viral genomic RNA into nucleocapsids and subsequently into virions is not completely understood. Phosphoprotein (P) and nucleoprotein (NP) interactions link NP-RNA complexes with P-L (polymerase) complexes to form viral nucleocapsids. The nucleocapsid then interacts with the viral matrix protein, leading to specific packaging of the nucleocapsid into the virion. A mammalian two-hybrid assay and confocal microscopy were used to demonstrate that Ebola virus VP35 and VP40 interact and colocalize in transfected cells. VP35 was packaged into budding virus-like particles (VLPs) as observed by protease protection assays. Moreover, VP40 and VP35 were sufficient for packaging an Ebola virus minigenome RNA into VLPs. Results from immunoprecipitation-reverse transcriptase PCR experiments suggest that VP35 confers specificity of the nucleocapsid for viral genomic RNA by direct VP35-RNA interactions.
authors.affiliation: Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, 3800 Spruce St., Philadelphia, PA 19104, USA.
authors.forename: Reed F
authors.initials: RF
authors.lastname: Johnson
authors.forename: Sarah E
authors.initials: SE
authors.lastname: McCarthy
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Godlewski
authors.forename: Ronald N
authors.initials: RN
authors.lastname: Harty
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D029043:Ebolavirus / Q000737:chemistry* / Q000648:ultrastructure; D016679:Genome, Viral; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000502:physiology*; D014709:Vero Cells; D014758:Viral Core Proteins / Q000502:physiology*; D019065:Virus Assembly / Q000502:physiology*
chemical_list: D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D014758:Viral Core Proteins; C084818:nucleoprotein VP35, Ebola virus; C084819:nucleoprotein VP40, Ebola virus