title: The autophagy elongation complex (ATG5-12/16L1) positively regulates HCV replication and is required for wild-type membranous web formation.
abstract: Hepatitis C virus (HCV) infection induces intracellular membrane rearrangements, thus forming a membranous web (MW) in which HCV replication and assembly occur. The HCV-induced MW is primarily composed of double membrane vesicles (DMVs) transfused by multi-membrane vesicles. The autophagy machinery has been proposed to participate in the formation of such vesicles. However, no clear evidence has been found linking autophagy to the formation of these DMVs. In this study, we evaluated the role of the autophagy elongation complex (ATG5-12/16L1) in HCV replication and MW formation. Using a dominant negative form of ATG12 and an siRNA approach, we demonstrated that the ATG5-12 conjugate, but not LC3-II formation, is crucial for efficient viral replication. Furthermore, purification of HCV MW revealed the presence of ATG5-12 and ATG16L1 along with HCV nonstructural proteins. Interestingly, LC3 was not recruited along with the elongation complex to the site of viral replication. Finally, inhibition of the elongation complex, but not LC3, greatly impaired the formation of the wild-type MW phenotype. To our knowledge, this study provides the first evidence of the involvement of autophagy proteins in the formation of wild-type MWs.
authors.affiliation: INRS-Institut Armand-Frappier, Institut National de la Recherche Scientifique, Laval, Canada.
authors.forename: Ahmed M
authors.initials: AM
authors.lastname: Fahmy
authors.affiliation: INRS-Institut Armand-Frappier, Institut National de la Recherche Scientifique, Laval, Canada.
authors.forename: Patrick
authors.initials: P
authors.lastname: Labonté
mesh_terms: D001343:Autophagy; D000071192:Autophagy-Related Protein 12 / Q000378:metabolism*; D000071187:Autophagy-Related Protein 5 / Q000378:metabolism*; D000071183:Autophagy-Related Proteins / Q000378:metabolism*; D045744:Cell Line, Tumor; D020868:Gene Silencing; D016174:Hepacivirus / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology*; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D010641:Phenotype; D011499:Protein Processing, Post-Translational; D014779:Virus Replication / Q000502:physiology*
chemical_list: C466624:ATG12 protein, human; C518959:ATG16L1 protein, human; C502778:ATG5 protein, human; D000071192:Autophagy-Related Protein 12; D000071187:Autophagy-Related Protein 5; D000071183:Autophagy-Related Proteins; C477012:MAP1LC3A protein, human; D008869:Microtubule-Associated Proteins