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Journal of Virology, December 2006, p. 11571-11578, Vol. 80, No. 23
0022-538X/06/$08.00+0     doi:10.1128/JVI.01717-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Hepatitis C Virus Entry Requires a Critical Postinternalization Step and Delivery to Early Endosomes via Clathrin-Coated Vesicles{triangledown}

Laurent Meertens, Claire Bertaux, and Tatjana Dragic*

Department of Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461

Received 8 August 2006/ Accepted 13 September 2006

Hepatitis C virus (HCV) is a major human pathogen associated with life-threatening liver disease. Entry into hepatocytes requires CD81 and a putative second receptor. In this study, we elucidated the postreceptor attachment stages of HCV entry using HCV pseudoparticles (HCVpp) as a model system. By means of dominant-negative mutants and short interfering RNAs of various cellular proteins, we showed that HCVpp enter via clathrin-coated vesicles and require delivery to early but not to late endosomes. However, the kinetics of HCV envelope glycoprotein-mediated fusion are delayed compared to those of other viruses that enter in early endosomes. Entry of HCVpp can be efficiently blocked by bafilomycin A1, which neutralizes the pH in early endosomes and impairs progression of endocytosis beyond this stage. However, low-pH exposure of bafilomycin A1-treated target cells does not induce entry of HCVpp at the plasma membrane or in the early stages of endocytosis. These observations indicate that, subsequent to internalization, HCVpp entry necessitates additional, low-pH-dependent interactions, modifications, or trafficking, and that these events are irreversibly disrupted by bafilomycin A1 treatment.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Albert Einstein College of Medicine, Jack 7 Pearl Resnick Campus, 1300 Morris Park Avenue, Golding B1, Bronx, NY 10461. Phone: (718) 430-3282. Fax: (718) 430-8711. E-mail: tdragic{at}aecom.yu.edu.

{triangledown} Published ahead of print on 27 September 2006.

Journal of Virology, December 2006, p. 11571-11578, Vol. 80, No. 23
0022-538X/06/$08.00+0     doi:10.1128/JVI.01717-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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