Subcellular localization of hepatitis C virus structural proteins in a cell culture system that efficiently replicates the virus.
Due to the recent development of a cell culture model, hepatitis C virus (HCV) can be efficiently propagated in cell culture. This allowed us to reinvestigate the subcellular localization of HCV structural proteins in the context of an infectious cycle. In agreement with previous reports, confocal immunofluorescence analysis of the subcellular localization of HCV structural proteins indicated that, in infected cells, the glycoprotein heterodimer is retained in the endoplasmic reticulum. However, in contrast to other studies, the glycoprotein heterodimer did not accumulate in other intracellular compartments or at the plasma membrane. As previously reported, an association between the capsid protein and lipid droplets was also observed. In addition, a fraction of labeling was consistent with the capsid protein being localized in a membranous compartment that is associated with the lipid droplets. However, in contrast to previous reports, the capsid protein was not found in the nucleus or in association with mitochondria or other well-defined intracellular compartments. Surprisingly, no colocalization was observed between the glycoprotein heterodimer and the capsid protein in infected cells. Electron microscopy analyses allowed us to identify a membrane alteration similar to the previously reported "membranous web." However, no virus-like particles were found in this type of structure. In addition, dense elements compatible with the size and shape of a viral particle were seldom observed in infected cells. In conclusion, the cell culture system for HCV allowed us for the first time to characterize the subcellular localization of HCV structural proteins in the context an infectious cycle.
Rouillé
Yves
Y
CNRS-UPR2511, Institut de Biologie de Lille, 1 Rue Calmette, BP447, 59021 Lille Cedex, France.
Helle
François
F
Delgrange
David
D
Roingeard
Philippe
P
Voisset
Cécile
C
Blanchard
Emmanuelle
E
Belouzard
Sandrine
S
McKeating
Jane
J
Patel
Arvind H
AH
Maertens
Geert
G
Wakita
Takaji
T
Wychowski
Czeslaw
C
Dubuisson
Jean
J
D036022:Capsid Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D019281:Dimerization; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000378:metabolism; D016174:Hepacivirus / Q000378:metabolism / Q000472:pathogenicity*; D006801:Humans; D008055:Lipids; D018613:Microscopy, Confocal; D013347:Subcellular Fractions / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000378:metabolism; D015678:Viral Structural Proteins / Q000378:metabolism*; D014779:Virus Replication
D036022:Capsid Proteins; D006023:Glycoproteins; D008055:Lipids; D014759:Viral Envelope Proteins; D015678:Viral Structural Proteins