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Efficient Initiation of HCV RNA Replication in Cell Culture
Keril J. Blight,1Alexander A. Kolykhalov,1Charles M. Rice12*
Hepatitis C virus (HCV) infection is a global
health problem affecting an estimated 170 million individuals
worldwide. Wereport the identification of multiple independent
adaptive mutationsthat cluster in the HCV nonstructural protein NS5A
and conferincreased replicative ability in vitro. Among these adaptive
mutationswere a single amino acid substitution that allowed HCV RNA
replicationin 10% of transfected hepatoma cells and a deletion of 47 aminoacids encompassing the interferon (IFN) sensitivity determiningregion (ISDR). Independent of the ISDR, IFN- rapidly inhibitedHCV
RNA replication in vitro. This work establishes a robust,cell-based
system for genetic and functional analyses of HCV replication.
1 Department of Molecular Microbiology,
Washington University School of Medicine, 660 South Euclid Avenue, St.
Louis, MO 63110-1093, USA.
2 Center for the Study
of Hepatitis C, Rockefeller University, Box 64, 1230 York Avenue, New
York, NY 10021, USA.
*
To whom correspondence should be addressed. E-mail:
ricec{at}rockvax.rockefeller.edu
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281, 9833-9836
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Mutations Conferring Resistance to SCH6, a Novel Hepatitis C Virus NS3/4A Protease Inhibitor: REDUCED RNA REPLICATION FITNESS AND PARTIAL RESCUE BY SECOND-SITE MUTATIONS.
M. Yi, X. Tong, A. Skelton, R. Chase, T. Chen, A. Prongay, S. L. Bogen, A. K. Saksena, F. G. Njoroge, R. L. Veselenak, et al. (2006)
J. Biol. Chem.
281, 8205-8215
|Abstract »|Full Text »|PDF »
From the Cover: Cell culture-grown hepatitis C virus is infectious in vivo and can be recultured in vitro.
B. D. Lindenbach, P. Meuleman, A. Ploss, T. Vanwolleghem, A. J. Syder, J. A. McKeating, R. E. Lanford, S. M. Feinstone, M. E. Major, G. Leroux-Roels, et al. (2006)
PNAS
103, 3805-3809
|Abstract »|Full Text »|PDF »
A milestone for hepatitis C virus research: A virus generated in cell culture is fully viable in vivo.
J. Bukh and R. H. Purcell (2006)
PNAS
103, 3500-3501
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SCH 503034, a Mechanism-Based Inhibitor of Hepatitis C Virus NS3 Protease, Suppresses Polyprotein Maturation and Enhances the Antiviral Activity of Alpha Interferon in Replicon Cells.
B. A. Malcolm, R. Liu, F. Lahser, S. Agrawal, B. Belanger, N. Butkiewicz, R. Chase, F. Gheyas, A. Hart, D. Hesk, et al. (2006)
Antimicrob. Agents Chemother.
50, 1013-1020
|Abstract »|Full Text »|PDF »
Intrahepatic Hepatitis C Virus Replication Correlates with Chronic Hepatitis C Disease Severity In Vivo.
S. Pal, M. C. Shuhart, L. Thomassen, S. S. Emerson, T. Su, N. Feuerborn, J. Kae, and D. R. Gretch (2006)
J. Virol.
80, 2280-2290
|Abstract »|Full Text »|PDF »
From the Cover: Production of infectious genotype 1a hepatitis C virus (Hutchinson strain) in cultured human hepatoma cells.
M. Yi, R. A. Villanueva, D. L. Thomas, T. Wakita, and S. M. Lemon (2006)
PNAS
103, 2310-2315
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Structural and Biological Identification of Residues on the Surface of NS3 Helicase Required for Optimal Replication of the Hepatitis C Virus.
S. G. Mackintosh, J. Z. Lu, J. B. Jordan, M. K. Harrison, B. Sikora, S. D. Sharma, C. E. Cameron, K. D. Raney, and J. Sakon (2006)
J. Biol. Chem.
281, 3528-3535
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Insertion and deletion analyses identify regions of non-structural protein 5A of Hepatitis C virus that are dispensable for viral genome replication.
S. Liu, I. H. Ansari, S. C. Das, and A. K. Pattnaik (2006)
J. Gen. Virol.
87, 323-327
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Effect of Cell Growth on Hepatitis C Virus (HCV) Replication and a Mechanism of Cell Confluence-Based Inhibition of HCV RNA and Protein Expression.
Activation of V{gamma}9V{delta}2 T cells by non-peptidic antigens induces the inhibition of subgenomic HCV replication.
C. Agrati, T. Alonzi, R. De Santis, C. Castilletti, I. Abbate, M. R. Capobianchi, G. D'Offizi, F. Siepi, G. M. Fimia, M. Tripodi, et al. (2006)
Int. Immunol.
18, 11-18
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Upregulation of Protein Phosphatase 2Ac by Hepatitis C Virus Modulates NS3 Helicase Activity through Inhibition of Protein Arginine Methyltransferase 1.
F. H. T. Duong, V. Christen, J. M. Berke, S. H. Penna, D. Moradpour, and M. H. Heim (2005)
J. Virol.
79, 15342-15350
|Abstract »|Full Text »|PDF »
Signal Peptide Cleavage and Internal Targeting Signals Direct the Hepatitis C Virus p7 Protein to Distinct Intracellular Membranes.
S. Griffin, D. Clarke, C. McCormick, D. Rowlands, and M. Harris (2005)
J. Virol.
79, 15525-15536
|Abstract »|Full Text »|PDF »
High-Throughput Assays Using a Luciferase-Expressing Replicon, Virus-Like Particles, and Full-Length Virus for West Nile Virus Drug Discovery.
F. Puig-Basagoiti, T. S. Deas, P. Ren, M. Tilgner, D. M. Ferguson, and P.-Y. Shi (2005)
Antimicrob. Agents Chemother.
49, 4980-4988
|Abstract »|Full Text »|PDF »
Robust Production of Infectious Hepatitis C Virus (HCV) from Stably HCV cDNA-Transfected Human Hepatoma Cells.
Z. Cai, C. Zhang, K.-S. Chang, J. Jiang, B.-C. Ahn, T. Wakita, T. J. Liang, and G. Luo (2005)
J. Virol.
79, 13963-13973
|Abstract »|Full Text »|PDF »
Functional Analysis of RNA Binding by the Hepatitis C Virus RNA-dependent RNA Polymerase.
Y.-C. Kim, W. K. Russell, C. T. Ranjith-Kumar, M. Thomson, D. H. Russell, and C. C. Kao (2005)
J. Biol. Chem.
280, 38011-38019
|Abstract »|Full Text »|PDF »
Development and characterization of a transient-replication assay for the genotype 2a hepatitis C virus subgenomic replicon.
rapidly inhibited HCV
RNA replication in vitro. This work establishes a robust, cell-based
system for genetic and functional analyses of HCV replication.
