Reduction of the infectivity of hepatitis C virus pseudoparticles by incorporation of misfolded glycoproteins induced by glucosidase inhibitors.
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.
Chapel
Cynthia
C
INSERM, U871, Université Lyon 1, et IFR62 Laennec, Lyon, France.
Garcia
Céline
C
INSERM, U871, Université Lyon 1, et IFR62 Laennec, Lyon, France.
Bartosch
Birke
B
INSERM, U758, Ecole Normale Supérieure de Lyon, et IFR128 BioSciences Lyon-Gerland, Lyon, France.
Roingeard
Philippe
P
INSERM, ESPRI 3856, Université François Rabelais, Tours, France.
Zitzmann
Nicole
N
Glycobiology Institute, Department of Biochemistry, University of Oxford, Oxford, UK.
Cosset
François-Loïc
FL
INSERM, U758, Ecole Normale Supérieure de Lyon, et IFR128 BioSciences Lyon-Gerland, Lyon, France.
Dubuisson
Jean
J
CNRS-UPR2511, Institut de Biologie de Lille et Institut Pasteur de Lille, Lille, France.
Dwek
Raymond A
RA
Glycobiology Institute, Department of Biochemistry, University of Oxford, Oxford, UK.
Trépo
Christian
C
INSERM, U871, Université Lyon 1, et IFR62 Laennec, Lyon, France.
Zoulim
Fabien
F
INSERM, U871, Université Lyon 1, et IFR62 Laennec, Lyon, France.
Durantel
David
D
INSERM, U871, Université Lyon 1, et IFR62 Laennec, Lyon, France.
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
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