title: In vivo oligomerization and raft localization of Ebola virus protein VP40 during vesicular budding.
abstract: The matrix protein VP40 plays a critical role in Ebola virus assembly and budding, a process that utilizes specialized membrane domains known as lipid rafts. Previous studies with purified protein suggest a role for oligomerization of VP40 in this process. Here, we demonstrate VP40 oligomers in lipid rafts of mammalian cells, virus-like particles, and in the authentic Ebola virus. By mutagenesis, we identify several critical C-terminal sequences that regulate oligomerization at the plasma membrane, association with detergent-resistant membranes, and vesicular release of VP40, directly linking these phenomena. Furthermore, we demonstrate the active recruitment of TSG101 into lipid rafts by VP40. We also report the successful application of the biarsenic fluorophore, FlAsH, combined with a tetracysteine tag for imaging of Ebola VP40 in live cells.
authors.affiliation: Developmental Therapeutics Program, Target Structure Based Drug Discovery Group, Science Applications International Corporation, National Cancer Institute, Frederick, MD 21702-1201, USA.
authors.forename: Rekha G
authors.initials: RG
authors.lastname: Panchal
authors.forename: Gordon
authors.initials: G
authors.lastname: Ruthel
authors.forename: Tara A
authors.initials: TA
authors.lastname: Kenny
authors.forename: George H
authors.initials: GH
authors.lastname: Kallstrom
authors.forename: Douglas
authors.initials: D
authors.lastname: Lane
authors.forename: Shirin S
authors.initials: SS
authors.lastname: Badie
authors.forename: Limin
authors.initials: L
authors.lastname: Li
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 / Q000821:virology; D029043:Ebolavirus / Q000235:genetics; D006801:Humans; D007668:Kidney; D021962:Membrane Microdomains / Q000821:virology*; D018613:Microscopy, Confocal; D009698:Nucleoproteins / Q000032:analysis / Q000235:genetics*; D011994:Recombinant Proteins / Q000032:analysis; D014162:Transfection; D014758:Viral Core Proteins / Q000032:analysis / Q000235:genetics*
chemical_list: D009698:Nucleoproteins; D011994:Recombinant Proteins; D014758:Viral Core Proteins; C084819:nucleoprotein VP40, Ebola virus