HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 21, 22371-22376, May 21, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/21/22371    most recent M311120200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Date, T. Articles by Wakita, T. Search for Related Content PubMed PubMed Citation Articles by Date, T. Articles by Wakita, T. Social Bookmarking                      What's this?

Genotype 2a Hepatitis C Virus Subgenomic Replicon Can Replicate in HepG2 and IMY-N9 Cells^*

Tomoko Date, Takanobu Kato, Michiko Miyamoto, Zijiang Zhao¶, Kotaro Yasui, Masashi Mizokami, and Takaji Wakita||

From the Department of Microbiology, Tokyo Metropolitan Institute for Neuroscience, Tokyo 183-8526, Japan, the Department of Clinical Molecular Informative Medicine, Nagoya City University Graduate School of Medical Sciences, Nagoya 467-8601, Japan, and the ^¶Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052, People's Republic of China

A hepatitis C virus genotype 2a subgenomic replicon, JFH-1 replicon, was previously established using the consensus sequence of clone JFH-1 from a patient with fulminant hepatitis and, in a previous report, was indicated to replicate efficiently in Huh7. Here the replication of JFH-1 replicon was tested in HepG2, a human hepatocyte-derived cell line, and in IMY-N9, a cell line developed by fusing human hepatocytes and HepG2 cells. Following transfection with in vitro transcribed replicon RNA and selection by cultivation with G418, colonies formed in both cell lines although at efficiencies substantially lower than those of Huh7. The H2476L mutation identified in the Huh7 replicon in our previous study increased the colony formation efficiencies of the JFH-1 replicon in HepG2 and IMY-N9 cells. Higher amounts of replicon RNA were detected in IMY-N9 clones than in HepG2 clones by real time detection reverse transcription-PCR, and replicon RNA replication and viral protein expression were confirmed by Northern and Western blotting in isolated clones. Sequencing of replicon RNAs revealed that mutations found in hepatitis C virus-derived regions were not identical and that two of nine HepG2 clones and three of nine IMY-N9 clones had no or one synonymous mutation. This system with the JFH-1 replicon and three cell lines is useful not only for estimating the cellular factors affecting viral activity but also for clarifying the common gene response of the host.

Received for publication, October 9, 2003 , and in revised form, February 19, 2004.

^* This work was supported in part by a grant-in-aid for Scientific Research from the Japan Society for the Promotion of Science and the Ministry of Health, Labour, and Welfare of Japan, by a grant from Toray Industries, Inc., and by the Program for Promotion of Fundamental Studies in Health Sciences of the Organization for Pharmaceutical Safety and Research of Japan. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence should be addressed: Dept. of Microbiology, Tokyo Metropolitan Institute for Neuroscience, 2-6 Musashidai, Fuchu, Tokyo 183-8526, Japan. Tel.: 81-423-25-3881; Fax: 81-423-21-8678; E-mail: wakita{at}tmin.ac.jp.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				C. J. Mee, H. J. Harris, M. J. Farquhar, G. Wilson, G. Reynolds, C. Davis, S. C. D. van IJzendoorn, P. Balfe, and J. A. McKeating Polarization Restricts Hepatitis C Virus Entry into HepG2 Hepatoma Cells J. Virol., June 15, 2009; 83(12): 6211 - 6221. [Abstract] [Full Text] [PDF]

				R. Adair, A. H. Patel, L. Corless, S. Griffin, D. J. Rowlands, and C. J. McCormick Expression of hepatitis C virus (HCV) structural proteins in trans facilitates encapsidation and transmission of HCV subgenomic RNA J. Gen. Virol., April 1, 2009; 90(4): 833 - 842. [Abstract] [Full Text] [PDF]

				J. Chang, J.-T. Guo, D. Jiang, H. Guo, J. M. Taylor, and T. M. Block Liver-Specific MicroRNA miR-122 Enhances the Replication of Hepatitis C Virus in Nonhepatic Cells J. Virol., August 15, 2008; 82(16): 8215 - 8223. [Abstract] [Full Text] [PDF]

				T. L. Tellinghuisen, M. J. Evans, T. von Hahn, S. You, and C. M. Rice Studying Hepatitis C Virus: Making the Best of a Bad Virus J. Virol., September 1, 2007; 81(17): 8853 - 8867. [Full Text] [PDF]

				T. Kato, T. Matsumura, T. Heller, S. Saito, R. K. Sapp, K. Murthy, T. Wakita, and T. J. Liang Production of Infectious Hepatitis C Virus of Various Genotypes in Cell Cultures J. Virol., May 1, 2007; 81(9): 4405 - 4411. [Abstract] [Full Text] [PDF]

				C. A. Lazaro, M. Chang, W. Tang, J. Campbell, D. G. Sullivan, D. R. Gretch, L. Corey, R. W. Coombs, and N. Fausto Hepatitis C Virus Replication in Transfected and Serum-Infected Cultured Human Fetal Hepatocytes Am. J. Pathol., February 1, 2007; 170(2): 478 - 489. [Abstract] [Full Text] [PDF]

				G. Koutsoudakis, E. Herrmann, S. Kallis, R. Bartenschlager, and T. Pietschmann The Level of CD81 Cell Surface Expression Is a Key Determinant for Productive Entry of Hepatitis C Virus into Host Cells J. Virol., January 15, 2007; 81(2): 588 - 598. [Abstract] [Full Text] [PDF]

				S. B. Kapadia, H. Barth, T. Baumert, J. A. McKeating, and F. V. Chisari Initiation of Hepatitis C Virus Infection Is Dependent on Cholesterol and Cooperativity between CD81 and Scavenger Receptor B Type I J. Virol., January 1, 2007; 81(1): 374 - 383. [Abstract] [Full Text] [PDF]

				T. Kato, T. Date, M. Miyamoto, M. Sugiyama, Y. Tanaka, E. Orito, T. Ohno, K. Sugihara, I. Hasegawa, K. Fujiwara, et al. Detection of Anti-Hepatitis C Virus Effects of Interferon and Ribavirin by a Sensitive Replicon System J. Clin. Microbiol., November 1, 2005; 43(11): 5679 - 5684. [Abstract] [Full Text] [PDF]

				P. Targett-Adams and J. McLauchlan Development and characterization of a transient-replication assay for the genotype 2a hepatitis C virus subgenomic replicon J. Gen. Virol., November 1, 2005; 86(11): 3075 - 3080. [Abstract] [Full Text] [PDF]

				M. P. Windisch, M. Frese, A. Kaul, M. Trippler, V. Lohmann, and R. Bartenschlager Dissecting the Interferon-Induced Inhibition of Hepatitis C Virus Replication by Using a Novel Host Cell Line J. Virol., November 1, 2005; 79(21): 13778 - 13793. [Abstract] [Full Text] [PDF]

				B. D. Lindenbach, M. J. Evans, A. J. Syder, B. Wolk, T. L. Tellinghuisen, C. C. Liu, T. Maruyama, R. O. Hynes, D. R. Burton, J. A. McKeating, et al. Complete Replication of Hepatitis C Virus in Cell Culture Science, July 22, 2005; 309(5734): 623 - 626. [Abstract] [Full Text] [PDF]

				J. Zhong, P. Gastaminza, G. Cheng, S. Kapadia, T. Kato, D. R. Burton, S. F. Wieland, S. L. Uprichard, T. Wakita, and F. V. Chisari Robust hepatitis C virus infection in vitro PNAS, June 28, 2005; 102(26): 9294 - 9299. [Abstract] [Full Text] [PDF]

				T. Kato, T. Date, M. Miyamoto, Z. Zhao, M. Mizokami, and T. Wakita Nonhepatic Cell Lines HeLa and 293 Support Efficient Replication of the Hepatitis C Virus Genotype 2a Subgenomic Replicon J. Virol., January 1, 2005; 79(1): 592 - 596. [Abstract] [Full Text] [PDF]

				N. Kanazawa, M. Kurosaki, N. Sakamoto, N. Enomoto, Y. Itsui, T. Yamashiro, Y. Tanabe, S. Maekawa, M. Nakagawa, C.-H. Chen, et al. Regulation of Hepatitis C Virus Replication by Interferon Regulatory Factor 1 J. Virol., September 15, 2004; 78(18): 9713 - 9720. [Abstract] [Full Text] [PDF]