title: Reduction of the infectivity of hepatitis C virus pseudoparticles by incorporation of misfolded glycoproteins induced by glucosidase inhibitors.
abstract: Folding and assembly into complexes of some viral glycoproteins are exquisitely sensitive to endoplasmic reticulum (ER) alpha-glucosidase inhibition, which prevents the trimming of glucose from N-linked glycans. Derivatives of deoxynojirimycin (DNJ) iminosugars, which are potent alpha-glucosidase inhibitors, were shown to have antiviral activity against bovine viral diarrhea virus, a pestivirus related to hepatitis C virus (HCV). The aim of this study was to determine whether these inhibitors would affect HCV infectivity and to provide novel insights on their mechanism of action. The overall antiviral activity of glucosidase inhibitors was shown by using the two most relevant models currently available: the cell-culture model enabling complete replication of the HCV JFH1 strain in Huh7.5 cells, and infectious HCV pseudotyped particles (HCVpp) produced in HEK-293T cells that display functional E1-E2 glycoprotein complexes. By using the latter model, it is shown that the inhibition of alpha-glucosidases by iminosugars results in the misfolding and misassembly of HCV glycoprotein pre-budding complexes. This inhibition of the assembly of E1-E2 in the ER of transfected HEK-293T cells leads to a reduction in the incorporation of E1-E2 complexes into HCVpp. More importantly, it is demonstrated that the infectivity of HCVpp that are released under treatment is reduced and that this reduction in infectivity is due to the incorporation of misfolded envelope glycoproteins in secreted particles. These properties suggest the potential usefulness of DNJ derivatives in combating HCV infection.
authors: INSERM, U871, Université Lyon 1, et IFR62 Laennec, Lyon, France.
authors: Cynthia
authors: C
authors: Chapel
authors: INSERM, U871, Université Lyon 1, et IFR62 Laennec, Lyon, France.
authors: Céline
authors: C
authors: Garcia
authors: INSERM, U758, Ecole Normale Supérieure de Lyon, et IFR128 BioSciences Lyon-Gerland, Lyon, France.
authors: Birke
authors: B
authors: Bartosch
authors: INSERM, ESPRI 3856, Université François Rabelais, Tours, France.
authors: Philippe
authors: P
authors: Roingeard
authors: Glycobiology Institute, Department of Biochemistry, University of Oxford, Oxford, UK.
authors: Nicole
authors: N
authors: Zitzmann
authors: INSERM, U758, Ecole Normale Supérieure de Lyon, et IFR128 BioSciences Lyon-Gerland, Lyon, France.
authors: François-Loïc
authors: FL
authors: Cosset
authors: CNRS-UPR2511, Institut de Biologie de Lille et Institut Pasteur de Lille, Lille, France.
authors: Jean
authors: J
authors: Dubuisson
authors: Glycobiology Institute, Department of Biochemistry, University of Oxford, Oxford, UK.
authors: Raymond A
authors: RA
authors: Dwek
authors: INSERM, U871, Université Lyon 1, et IFR62 Laennec, Lyon, France.
authors: Christian
authors: C
authors: Trépo
authors: INSERM, U871, Université Lyon 1, et IFR62 Laennec, Lyon, France.
authors: Fabien
authors: F
authors: Zoulim
authors: INSERM, U871, Université Lyon 1, et IFR62 Laennec, Lyon, France.
authors: David
authors: D
authors: Durantel
mesh_terms: D017485:1-Deoxynojirimycin / Q000494:pharmacology; D000998:Antiviral Agents / Q000494:pharmacology; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005944:Glucosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D065089:Glycoside Hydrolase Inhibitors; D006031:Glycosylation; D016174:Hepacivirus / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D016253:Microscopy, Immunoelectron; D011485:Protein Binding / Q000187:drug effects; D017510:Protein Folding; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000378:metabolism*; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; C087035:E1 protein, Hepatitis C virus; D004791:Enzyme Inhibitors; D065089:Glycoside Hydrolase Inhibitors; D014759:Viral Envelope Proteins; C501633:deoxynojirimycine; C091015:glycoprotein E2, Hepatitis C virus; D017485:1-Deoxynojirimycin; D008156:Luciferases; D005944:Glucosamine