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Investigating the Role of Cellular Lipid Recognition in Trafficking and Viral Particle Formation by Ebola Virus Matrix Protein VP40
Author(s) -
Johnson Kristen,
Stahelin Robert
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.886.6
Subject(s) - vp40 , ebola virus , viral matrix protein , matrix (chemical analysis) , microbiology and biotechnology , virology , virus like particle , virus , chemistry , biology , biochemistry , gene , chromatography , recombinant dna
Virulent pathogens hijack host cell machinery during their infection scheme. Commonly, fundamental biochemical processes are discovered through investigation of pathogen infection, especially in the case of pathogen modulation of low abundance lipid species. Ebola virus (EBOV) is a highly virulent, filamentous, lipid‐enveloped virus that causes hemorrhagic fever and a fatality rate of 50‐90%. The limited understanding of the viral replication in host cells proves difficult for understanding how it spreads. EBOV only has seven genes in its genome, one of which‐VP40‐is essential for viral egress. VP40 can form virus like particles (VLPs) on its own at the plasma membrane (PM) in live cells and is an excellent model for studying EBOV spread. Objective: We are implementing several assays in live COS‐7 cells to decode the PM phospholipid specificity where viral particle formation occurs. Methods: Confocal Laser Scanning Microscopy (CLSM), Spinning Disk and Total Internal Reflection Microscopy (TIRF), VLP Collection. Results: VP40 PM localization can be blocked by elimination of phospholipid substrates through various assays in live COS‐7 cells. We have demonstrated that both PS and PI(4,5)P2 are important for VP40 localization and VLP formation at the PM in live cells. We are also investigating the implications on VLP budding as the result of a decrease in PM localization. Conclusions Phospholipid‐focused assays in live cells reveal VP40 lipid binding specificity at various cellular sites, other quantitative methods aid in decoding the dynamics and nature of these interactions. Financial Support: NIH AI081077