Palmitoylation mediates membrane association of hepatitis E virus ORF3 protein and is required for infectious particle secretion.

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State: Public
Version: Final published version
License: CC BY 4.0
Serval ID
serval:BIB_48912260B048
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Palmitoylation mediates membrane association of hepatitis E virus ORF3 protein and is required for infectious particle secretion.
Journal
PLoS pathogens
Author(s)
Gouttenoire J., Pollán A., Abrami L., Oechslin N., Mauron J., Matter M., Oppliger J., Szkolnicka D., Dao Thi V.L., van der Goot F.G., Moradpour D.
ISSN
1553-7374 (Electronic)
ISSN-L
1553-7366
Publication state
Published
Issued date
12/2018
Peer-reviewed
Oui
Volume
14
Number
12
Pages
e1007471
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Abstract
Hepatitis E virus (HEV) is a positive-strand RNA virus encoding 3 open reading frames (ORF). HEV ORF3 protein is a small, hitherto poorly characterized protein involved in viral particle secretion and possibly other functions. Here, we show that HEV ORF3 protein forms membrane-associated oligomers. Immunoblot analyses of ORF3 protein expressed in cell-free vs. cellular systems suggested a posttranslational modification. Further analyses revealed that HEV ORF3 protein is palmitoylated at cysteine residues in its N-terminal region, as corroborated by 3H-palmitate labeling, the investigation of cysteine-to-alanine substitution mutants and treatment with the palmitoylation inhibitor 2-bromopalmitate (2-BP). Abrogation of palmitoylation by site-directed mutagenesis or 2-BP treatment altered the subcellular localization of ORF3 protein, reduced the stability of the protein and strongly impaired the secretion of infectious particles. Moreover, selective membrane permeabilization coupled with immunofluorescence microscopy revealed that HEV ORF3 protein is entirely exposed to the cytosolic side of the membrane, allowing to propose a model for its membrane topology and interactions required in the viral life cycle. In conclusion, palmitoylation determines the subcellular localization, membrane topology and function of HEV ORF3 protein in the HEV life cycle.
Keywords
Cell Line, Hepatitis E/virology, Hepatitis E virus/pathogenicity, Humans, Lipoylation, Viral Proteins/metabolism, Virus Release/physiology
Pubmed
Web of science
Open Access
Yes
Create date
05/01/2019 16:14
Last modification date
22/08/2019 13:13
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