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Journal of Virology, March 2006, p. 2832-2841, Vol. 80, No. 6
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.6.2832-2841.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Subcellular Localization of Hepatitis C Virus Structural Proteins in a Cell Culture System That Efficiently Replicates the Virus

Yves Rouillé,1 François Helle,1 David Delgrange,1 Philippe Roingeard,2 Cécile Voisset,1 Emmanuelle Blanchard,1 Sandrine Belouzard,1 Jane McKeating,3 Arvind H. Patel,4 Geert Maertens,5 Takaji Wakita,6 Czeslaw Wychowski,1,{dagger} and Jean Dubuisson1*,{dagger}

CNRS-UPR2511, Institut de Biologie de Lille-Institut Pasteur de Lille, Lille, France,1 INSERM-ESPRI3856, François Rabelais University, Tours, France,2 Institute of Biomedical Research, University of Birmingham, Birmingham, United Kingdom,3 MRC Virology Unit, Institute of Virology, Glasgow, United Kingdom,4 Innogenetics, Ghent, Belgium,5 Department of Microbiology, Tokyo Metropolitan Institute for Neuroscience, Tokyo, Japan6

Received 23 September 2005/ Accepted 23 December 2005

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.

* Corresponding author. Mailing address: Unité Hépatite C, CNRS-UPR2511, Institut de Biologie de Lille, 1 Rue Calmette, BP447, 59021 Lille Cedex, France. Phone: (33) 3 20 87 11 60. Fax: (33) 3 20 87 12 01. E-mail: jean.dubuisson{at}ibl.fr.

{dagger} C.W. and J.D. contributed equally to this study.

Journal of Virology, March 2006, p. 2832-2841, Vol. 80, No. 6
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.6.2832-2841.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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