This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Yi, M.
Right arrow Articles by Lemon, S. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yi, M.
Right arrow Articles by Lemon, S. M.

 Previous Article  |  Next Article 

Journal of Virology, January 2007, p. 629-638, Vol. 81, No. 2
0022-538X/07/$08.00+0     doi:10.1128/JVI.01890-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Compensatory Mutations in E1, p7, NS2, and NS3 Enhance Yields of Cell Culture-Infectious Intergenotypic Chimeric Hepatitis C Virus{triangledown}

MinKyung Yi, Yinghong Ma, Jeremy Yates, and Stanley M. Lemon*

Center for Hepatitis Research, Institute for Human Infections and Immunity and Department of Microbiology and Immunology, University of Texas Medical Branch at Galveston, Galveston, Texas 77555-1019

Received 30 August 2006/ Accepted 22 October 2006

There is little understanding of mechanisms underlying the assembly and release of infectious hepatitis C virus (HCV) from cultured cells. Cells transfected with synthetic genomic RNA from a unique genotype 2a virus (JFH1) produce high titers of virus, while virus yields are much lower with a prototype genotype 1a RNA containing multiple cell culture-adaptive mutations (H77S). To characterize the basis for this difference in infectious particle production, we constructed chimeric genomes encoding the structural proteins of H77S within the background of JFH1. RNAs encoding polyproteins fused at the NS2/NS3 junction ("H-NS2/NS3-J") and at a site of natural, intergenotypic recombination within NS2 ["H-(NS2)-J"] produced infectious virus. In contrast, no virus was produced by a chimera fused at the p7-NS2 junction. Chimera H-NS2/NS3-J virus (vH-NS2/NS3-J) recovered from transfected cultures contained compensatory mutations in E1 and NS3 that were essential for the production of infectious virus, while yields of infectious vH-(NS2)-J were enhanced by mutations within p7 and NS2. These compensatory mutations were chimera specific and did not enhance viral RNA replication or polyprotein processing; thus, they likely compensate for incompatibilities between proteins of different genotypes at sites of interactions essential for virus assembly and/or release. Mutations in p7 and NS2 acted additively and increased the specific infectivity of vH-(NS2)-J particles, while having less impact on the numbers of particles released. We conclude that interactions between NS2 and E1 and p7 as well as between NS2 and NS3 are essential for virus assembly and/or release and that each of these viral proteins plays an important role in this process.

* Corresponding author. Mailing address: Center for Hepatitis Research, Institute for Human Infections and Immunity, The University of Texas Medical Branch at Galveston, 301 University Boulevard, Galveston, TX 77555-1019. Phone: (409) 747-7048. Fax: (409) 747-7030. E-mail: smlemon{at}utmb.edu.

{triangledown} Published ahead of print on 1 November 2006.

Journal of Virology, January 2007, p. 629-638, Vol. 81, No. 2
0022-538X/07/$08.00+0     doi:10.1128/JVI.01890-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Brohm, C., Steinmann, E., Friesland, M., Lorenz, I. C., Patel, A., Penin, F., Bartenschlager, R., Pietschmann, T. (2009). Characterization of Determinants Important for Hepatitis C Virus p7 Function in Morphogenesis by Using trans-Complementation. J. Virol. 83: 11682-11693 [Abstract] [Full Text]  
  • Pan, K.-L., Lee, J.-C., Sung, H.-W., Chang, T.-Y., Hsu, J. T.-A. (2009). Development of NS3/4A Protease-Based Reporter Assay Suitable for Efficiently Assessing Hepatitis C Virus Infection. Antimicrob. Agents Chemother. 53: 4825-4834 [Abstract] [Full Text]  
  • Wang, N., Liang, Y., Devaraj, S., Wang, J., Lemon, S. M., Li, K. (2009). Toll-Like Receptor 3 Mediates Establishment of an Antiviral State against Hepatitis C Virus in Hepatoma Cells. J. Virol. 83: 9824-9834 [Abstract] [Full Text]  
  • Phan, T., Beran, R. K. F., Peters, C., Lorenz, I. C., Lindenbach, B. D. (2009). Hepatitis C Virus NS2 Protein Contributes to Virus Particle Assembly via Opposing Epistatic Interactions with the E1-E2 Glycoprotein and NS3-NS4A Enzyme Complexes. J. Virol. 83: 8379-8395 [Abstract] [Full Text]  
  • McGivern, D. R., Villanueva, R. A., Chinnaswamy, S., Kao, C. C., Lemon, S. M. (2009). Impaired Replication of Hepatitis C Virus Containing Mutations in a Conserved NS5B Retinoblastoma Protein-Binding Motif. J. Virol. 83: 7422-7433 [Abstract] [Full Text]  
  • Bungyoku, Y., Shoji, I., Makine, T., Adachi, T., Hayashida, K., Nagano-Fujii, M., Ide, Y.-H., Deng, L., Hotta, H. (2009). Efficient production of infectious hepatitis C virus with adaptive mutations in cultured hepatoma cells. J. Gen. Virol. 90: 1681-1691 [Abstract] [Full Text]  
  • Welbourn, S., Jirasko, V., Breton, V., Reiss, S., Penin, F., Bartenschlager, R., Pause, A. (2009). Investigation of a role for lysine residues in non-structural proteins 2 and 2/3 of the hepatitis C virus for their degradation and virus assembly. J. Gen. Virol. 90: 1071-1080 [Abstract] [Full Text]  
  • Adair, R., Patel, A. H., Corless, L., Griffin, S., Rowlands, D. J., McCormick, C. J. (2009). Expression of hepatitis C virus (HCV) structural proteins in trans facilitates encapsidation and transmission of HCV subgenomic RNA. J. Gen. Virol. 90: 833-842 [Abstract] [Full Text]  
  • Beran, R. K. F., Lindenbach, B. D., Pyle, A. M. (2009). The NS4A Protein of Hepatitis C Virus Promotes RNA-Coupled ATP Hydrolysis by the NS3 Helicase. J. Virol. 83: 3268-3275 [Abstract] [Full Text]  
  • Gal-Tanamy, M., Keck, Z.-Y., Yi, M., McKeating, J. A., Patel, A. H., Foung, S. K. H., Lemon, S. M. (2008). In vitro selection of a neutralization-resistant hepatitis C virus escape mutant. Proc. Natl. Acad. Sci. USA 105: 19450-19455 [Abstract] [Full Text]  
  • Beran, R. K. F., Pyle, A. M. (2008). Hepatitis C Viral NS3-4A Protease Activity Is Enhanced by the NS3 Helicase. J. Biol. Chem. 283: 29929-29937 [Abstract] [Full Text]  
  • Jirasko, V., Montserret, R., Appel, N., Janvier, A., Eustachi, L., Brohm, C., Steinmann, E., Pietschmann, T., Penin, F., Bartenschlager, R. (2008). Structural and Functional Characterization of Nonstructural Protein 2 for Its Role in Hepatitis C Virus Assembly. J. Biol. Chem. 283: 28546-28562 [Abstract] [Full Text]  
  • Ma, Y., Yates, J., Liang, Y., Lemon, S. M., Yi, M. (2008). NS3 Helicase Domains Involved in Infectious Intracellular Hepatitis C Virus Particle Assembly. J. Virol. 82: 7624-7639 [Abstract] [Full Text]  
  • Steinmann, E., Brohm, C., Kallis, S., Bartenschlager, R., Pietschmann, T. (2008). Efficient trans-Encapsidation of Hepatitis C Virus RNAs into Infectious Virus-Like Particles. J. Virol. 82: 7034-7046 [Abstract] [Full Text]  
  • Campo, D. S., Dimitrova, Z., Mitchell, R. J., Lara, J., Khudyakov, Y. (2008). Coordinated evolution of the hepatitis C virus. Proc. Natl. Acad. Sci. USA 105: 9685-9690 [Abstract] [Full Text]  
  • Keck, Z.-Y., Li, T.-K., Xia, J., Gal-Tanamy, M., Olson, O., Li, S. H., Patel, A. H., Ball, J. K., Lemon, S. M., Foung, S. K. H. (2008). Definition of a Conserved Immunodominant Domain on Hepatitis C Virus E2 Glycoprotein by Neutralizing Human Monoclonal Antibodies. J. Virol. 82: 6061-6066 [Abstract] [Full Text]  
  • Oem, J.-K., Jackel-Cram, C., Li, Y.-P., Zhou, Y., Zhong, J., Shimano, H., Babiuk, L. A., Liu, Q. (2008). Activation of sterol regulatory element-binding protein 1c and fatty acid synthase transcription by hepatitis C virus non-structural protein 2. J. Gen. Virol. 89: 1225-1230 [Abstract] [Full Text]  
  • Russell, R. S., Meunier, J.-C., Takikawa, S., Faulk, K., Engle, R. E., Bukh, J., Purcell, R. H., Emerson, S. U. (2008). Advantages of a single-cycle production assay to study cell culture-adaptive mutations of hepatitis C virus. Proc. Natl. Acad. Sci. USA 105: 4370-4375 [Abstract] [Full Text]  
  • Zhang, Y., Weady, P., Duggal, R., Hao, W. (2008). Novel Chimeric Genotype 1b/2a Hepatitis C Virus Suitable for High-Throughput Screening. Antimicrob. Agents Chemother. 52: 666-674 [Abstract] [Full Text]  
  • Scheel, T. K. H., Gottwein, J. M., Jensen, T. B., Prentoe, J. C., Hoegh, A. M., Alter, H. J., Eugen-Olsen, J., Bukh, J. (2008). Development of JFH1-based cell culture systems for hepatitis C virus genotype 4a and evidence for cross-genotype neutralization. Proc. Natl. Acad. Sci. USA 105: 997-1002 [Abstract] [Full Text]  
  • Kaul, A., Woerz, I., Meuleman, P., Leroux-Roels, G., Bartenschlager, R. (2007). Cell Culture Adaptation of Hepatitis C Virus and In Vivo Viability of an Adapted Variant. J. Virol. 81: 13168-13179 [Abstract] [Full Text]  
  • Beran, R. K. F., Serebrov, V., Pyle, A. M. (2007). The Serine Protease Domain of Hepatitis C Viral NS3 Activates RNA Helicase Activity by Promoting the Binding of RNA Substrate. J. Biol. Chem. 282: 34913-34920 [Abstract] [Full Text]  
  • Murray, C. L., Jones, C. T., Tassello, J., Rice, C. M. (2007). Alanine Scanning of the Hepatitis C Virus Core Protein Reveals Numerous Residues Essential for Production of Infectious Virus. J. Virol. 81: 10220-10231 [Abstract] [Full Text]  
  • Jones, C. T., Murray, C. L., Eastman, D. K., Tassello, J., Rice, C. M. (2007). Hepatitis C Virus p7 and NS2 Proteins Are Essential for Production of Infectious Virus. J. Virol. 81: 8374-8383 [Abstract] [Full Text]  
  • Ishida, H., Li, K., Yi, M., Lemon, S. M. (2007). 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. J. Biol. Chem. 282: 11836-11848 [Abstract] [Full Text]  
  • Tscherne, D. M., Evans, M. J., von Hahn, T., Jones, C. T., Stamataki, Z., McKeating, J. A., Lindenbach, B. D., Rice, C. M. (2007). Superinfection Exclusion in Cells Infected with Hepatitis C Virus. J. Virol. 81: 3693-3703 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»