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Science 2 July 1999:
Vol. 285. no. 5424, pp. 110 - 113
DOI: 10.1126/science.285.5424.110
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Reports

Replication of Subgenomic Hepatitis C Virus RNAs in a Hepatoma Cell Line

V. Lohmann, 1 F. Körner, 1 J.-O. Koch, 1 U. Herian, 1 L. Theilmann, 2 R. Bartenschlager 1*

An estimated 170 million persons worldwide are infected with hepatitis C virus (HCV), a major cause of chronic liver disease. Despite increasing knowledge of genome structure and individual viral proteins, studies on virus replication and pathogenesis have been hampered by the lack of reliable and efficient cell culture systems. A full-length consensus genome was cloned from viral RNA isolated from an infected human liver and used to construct subgenomic selectable replicons. Upon transfection into a human hepatoma cell line, these RNAs were found to replicate to high levels, permitting metabolic radiolabeling of viral RNA and proteins. This work defines the structure of HCV replicons functional in cell culture and provides the basis for a long-sought cellular system that should allow detailed molecular studies of HCV and the development of antiviral drugs.

1 Institute for Virology, Johannes-Gutenberg University Mainz, Obere Zahlbacher Strasse 67, 55131 Mainz, Germany.
2 Städtisches Klinikum Pforzheim, Medizinische Klinik II, 75116 Pforzheim, Germany.
*   To whom correspondence should be addressed. E-mail: bartnsch{at}mail.uni-mainz.de

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J. Virol. 81, 8374-8383
   Abstract »    Full Text »    PDF »
Phenotypic and Structural Analyses of Hepatitis C Virus NS3 Protease Arg155 Variants: SENSITIVITY TO TELAPREVIR (VX-950) AND INTERFERON {alpha}.
Y. Zhou, U. Muh, B. L. Hanzelka, D. J. Bartels, Y. Wei, B. G. Rao, D. L. Brennan, A. M. Tigges, L. Swenson, A. D. Kwong, et al. (2007)
J. Biol. Chem. 282, 22619-22628
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The NS3 Helicase and NS5B-to-3'X Regions Are Important for Efficient Hepatitis C Virus Strain JFH-1 Replication in Huh7 Cells.
A. Murayama, T. Date, K. Morikawa, D. Akazawa, M. Miyamoto, M. Kaga, K. Ishii, T. Suzuki, T. Kato, M. Mizokami, et al. (2007)
J. Virol. 81, 8030-8040
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Cellular cofactors affecting hepatitis C virus infection and replication.
G. Randall, M. Panis, J. D. Cooper, T. L. Tellinghuisen, K. E. Sukhodolets, S. Pfeffer, M. Landthaler, P. Landgraf, S. Kan, B. D. Lindenbach, et al. (2007)
PNAS 104, 12884-12889
   Abstract »    Full Text »    PDF »
Binding Dynamics of Hepatitis C Virus' NS5A Amphipathic Peptide to Cell and Model Membranes.
N.-J. Cho, K. H. Cheong, C. Lee, C. W. Frank, and J. S. Glenn (2007)
J. Virol. 81, 6682-6689
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Comprehensive Analysis of the Effects of Ordinary Nutrients on Hepatitis C Virus RNA Replication in Cell Culture.
M. Yano, M. Ikeda, K.-i. Abe, H. Dansako, S. Ohkoshi, Y. Aoyagi, and N. Kato (2007)
Antimicrob. Agents Chemother. 51, 2016-2027
   Abstract »    Full Text »    PDF »
Allelic Variation in the Hepatitis C Virus NS4B Protein Dramatically Influences RNA Replication.
K. J. Blight (2007)
J. Virol. 81, 5724-5736
   Abstract »    Full Text »    PDF »
Characterization of Hepatitis C Virus Subgenomic Replicon Resistance to Cyclosporine In Vitro.
J. M. Robida, H. B. Nelson, Z. Liu, and H. Tang (2007)
J. Virol. 81, 5829-5840
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Functional Interaction of Hepatitis C Virus NS5B with Nucleolin GAR Domain.
T. Kusakawa, T. Shimakami, S. Kaneko, K. Yoshioka, and S. Murakami (2007)
J. Biochem. 141, 917-927
   Abstract »    Full Text »    PDF »
Identification of Determinants Involved in Initiation of Hepatitis C Virus RNA Synthesis by Using Intergenotypic Replicase Chimeras.
M. Binder, D. Quinkert, O. Bochkarova, R. Klein, N. Kezmic, R. Bartenschlager, and V. Lohmann (2007)
J. Virol. 81, 5270-5283
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Analysis of Hepatitis C Virus Superinfection Exclusion by Using Novel Fluorochrome Gene-Tagged Viral Genomes.
T. Schaller, N. Appel, G. Koutsoudakis, S. Kallis, V. Lohmann, T. Pietschmann, and R. Bartenschlager (2007)
J. Virol. 81, 4591-4603
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p21-activated Kinase 1 Is Activated through the Mammalian Target of Rapamycin/p70 S6 Kinase Pathway and Regulates the Replication of Hepatitis C Virus in Human Hepatoma Cells.
H. Ishida, K. Li, M. Yi, and S. M. Lemon (2007)
J. Biol. Chem. 282, 11836-11848
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An RNA-Binding Protein, hnRNP A1, and a Scaffold Protein, Septin 6, Facilitate Hepatitis C Virus Replication.
C. S. Kim, S. K. Seol, O.-K. Song, J. H. Park, and S. K. Jang (2007)
J. Virol. 81, 3852-3865
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From the Cover: Hepatitis C virus production by human hepatocytes dependent on assembly and secretion of very low-density lipoproteins.
H. Huang, F. Sun, D. M. Owen, W. Li, Y. Chen, M. Gale Jr., and J. Ye (2007)
PNAS 104, 5848-5853
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Reduction of the infectivity of hepatitis C virus pseudoparticles by incorporation of misfolded glycoproteins induced by glucosidase inhibitors.
C. Chapel, C. Garcia, B. Bartosch, P. Roingeard, N. Zitzmann, F.-L. Cosset, J. Dubuisson, R. A. Dwek, C. Trepo, F. Zoulim, et al. (2007)
J. Gen. Virol. 88, 1133-1143
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Upregulation of Indoleamine 2,3-Dioxygenase in Hepatitis C Virus Infection.
E. Larrea, J. I. Riezu-Boj, L. Gil-Guerrero, N. Casares, R. Aldabe, P. Sarobe, M. P. Civeira, J. L. Heeney, C. Rollier, B. Verstrepen, et al. (2007)
J. Virol. 81, 3662-3666
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Evidence for a functional RNA element in the hepatitis C virus core gene.
L. K. McMullan, A. Grakoui, M. J. Evans, K. Mihalik, M. Puig, A. D. Branch, S. M. Feinstone, and C. M. Rice (2007)
PNAS 104, 2879-2884
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Selective Repression of Translation by the Brome Mosaic Virus 1a RNA Replication Protein.
G. Yi, K. Gopinath, and C. C. Kao (2007)
J. Virol. 81, 1601-1609
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Mobility analysis of an NS5A-GFP fusion protein in cells actively replicating hepatitis C virus subgenomic RNA.
D. M. Jones, S. N. Gretton, J. McLauchlan, and P. Targett-Adams (2007)
J. Gen. Virol. 88, 470-475
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Hepatitis C Virus Replication in Transfected and Serum-Infected Cultured Human Fetal Hepatocytes.
C. A. Lazaro, M. Chang, W. Tang, J. Campbell, D. G. Sullivan, D. R. Gretch, L. Corey, R. W. Coombs, and N. Fausto (2007)
Am. J. Pathol. 170, 478-489
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