Open AccessIn vivo oligomerization and raft localization of Ebola virus protein VP40 during vesicular budding
Author(s) -
Rekha G. Panchal,
Gordon Ruthel,
Tara Kenny,
George Kallstrom,
Douglas Lane,
Shirin S. Badie,
Limin Liu,
Sina Bavari,
M. Javad Aman
Publication year - 2003
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.2533915100
Subject(s) - vp40 , ebola virus , viral matrix protein , lipid raft , microbiology and biotechnology , filoviridae , biology , chemistry , virus , virology , signal transduction , paramyxoviridae , viral disease
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.