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Journal of Virology, May 2003, p. 5487-5492, Vol. 77, No. 9
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.9.5487-5492.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification of the Hepatitis C Virus RNA Replication Complex in Huh-7 Cells Harboring Subgenomic Replicons

Rainer Gosert,1,2 Denise Egger,2 Volker Lohmann,3 Ralf Bartenschlager,3 Hubert E. Blum,1 Kurt Bienz,2 and Darius Moradpour1*

Department of Medicine II, University of Freiburg, D-79106 Freiburg,1 Department of Molecular Virology, University of Heidelberg, D-69120 Heidelberg, Germany,3 Institute for Medical Microbiology, University of Basel, CH-4003 Basel, Switzerland2

Received 18 October 2002/ Accepted 5 February 2003

Formation of a membrane-associated replication complex, composed of viral proteins, replicating RNA, and altered cellular membranes, is a characteristic feature of plus-strand RNA viruses. Here, we demonstrate the presence of a specific membrane alteration, designated the membranous web, that contains hepatitis C virus (HCV) nonstructural proteins, as well as viral plus-strand RNA, in Huh-7 cells harboring autonomously replicating subgenomic HCV RNAs. Metabolic labeling with 5-bromouridine 5'-triphosphate in the presence of actinomycin D revealed that the membranous web is the site of viral RNA synthesis and therefore represents the replication complex of HCV.

* Corresponding author. Mailing address: Department of Medicine II, University of Freiburg, D-79106 Freiburg, Germany. Phone: 49 761 270 3510. Fax: 49 761 270 3610. E-mail: Darius.Moradpour{at}uni-freiburg.de.

Journal of Virology, May 2003, p. 5487-5492, Vol. 77, No. 9
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.9.5487-5492.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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