title: Involvement of vacuolar protein sorting pathway in Ebola virus release independent of TSG101 interaction.
abstract: Budding of Ebola virus (EBOV) particles from the plasma membrane of infected cells requires viral and host proteins. EBOV virus matrix protein VP40 recruits TSG101, an ESCRT-1 (host cell endosomal sorting complex required for transport-1) complex protein in the vacuolar protein sorting (vps) pathway, to the plasma membrane during budding. Involvement of other vps proteins in EBOV budding has not been established. Therefore, we used VP40 deletion analysis, virus-like particle-release assays, and confocal microscopy to investigate the potential role of ESCRT-1 proteins VPS4, VPS28, and VPS37B in EBOV budding. We found that VP40 could redirect each protein from endosomes to the cell surface independently of TSG101 interaction. A lack of VPS4 adenosine triphosphatase activity reduced budding by up to 80%. Inhibition of VPS4 gene expression by use of phosphorodiamidite morpholino antisense oligonucleotides protected mice from lethal EBOV infection. These data show that EBOV can use vps proteins independently of TSG101 for budding and reveal VPS4 as a potential target for filovirus therapeutics.
authors.affiliation: United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD 21704, USA.
authors.forename: Lynn S
authors.initials: LS
authors.lastname: Silvestri
authors.forename: Gordon
authors.initials: G
authors.lastname: Ruthel
authors.forename: George
authors.initials: G
authors.lastname: Kallstrom
authors.forename: Kelly L
authors.initials: KL
authors.lastname: Warfield
authors.forename: Dana L
authors.initials: DL
authors.lastname: Swenson
authors.forename: Timothy
authors.initials: T
authors.lastname: Nelle
authors.forename: Patrick L
authors.initials: PL
authors.lastname: Iversen
authors.forename: Sina
authors.initials: S
authors.lastname: Bavari
authors.forename: M Javad
authors.initials: MJ
authors.lastname: Aman
mesh_terms: D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology / Q000821:virology; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000502:physiology*; D029043:Ebolavirus / Q000502:physiology*; D056827:Endosomal Sorting Complexes Required for Transport; D006801:Humans; D007668:Kidney; D018613:Microscopy, Confocal; D016296:Mutagenesis; D009838:Oligodeoxyribonucleotides; D021381:Protein Transport / Q000502:physiology; D014157:Transcription Factors / Q000502:physiology*; D014617:Vacuoles / Q000502:physiology*; D014764:Viral Proteins / Q000502:physiology*; D000099177:Tsg101 Protein
chemical_list: D017931:DNA Primers; D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D009838:Oligodeoxyribonucleotides; D014157:Transcription Factors; D014764:Viral Proteins; D000099177:Tsg101 Protein