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 Binder, M.
Right arrow Articles by Lohmann, V.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Binder, M.
Right arrow Articles by Lohmann, V.

 Previous Article  |  Next Article 

Journal of Virology, May 2007, p. 5270-5283, Vol. 81, No. 10
0022-538X/07/$08.00+0     doi:10.1128/JVI.00032-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Identification of Determinants Involved in Initiation of Hepatitis C Virus RNA Synthesis by Using Intergenotypic Replicase Chimeras{triangledown}

Marco Binder,{dagger} Doris Quinkert,{dagger} Olga Bochkarova, Rahel Klein, Nikolina Kezmic, Ralf Bartenschlager, and Volker Lohmann*

University of Heidelberg, Department of Molecular Virology, Im Neuenheimer Feld 345, 69120 Heidelberg, Germany

Received 5 January 2007/ Accepted 26 February 2007

The 5' nontranslated region (NTR) and the X tail in the 3' NTR are the least variable parts of the hepatitis C virus (HCV) genome and play an important role in the initiation of RNA synthesis. By using subgenomic replicons of the HCV isolates Con1 (genotype 1) and JFH1 (genotype 2), we characterized the genotype specificities of the replication signals contained in the NTRs. The replacement of the JFH1 5' NTR and X tail with the corresponding Con1 sequence resulted in a significant decrease in replication efficiency. Exchange of the X tail specifically reduced negative-strand synthesis, whereas substitution of the 5' NTR impaired the generation of progeny positive strands. In search for the proteins involved in the recognition of genotype-specific initiation signals, we analyzed recombinant nonstructural protein 5B (NS5B) RNA polymerases of both isolates and found some genotype-specific template preference for the 3' end of positive-strand RNA in vitro. To further address genotype specificity, we constructed a series of intergenotypic replicon chimeras. When combining NS3 to NS5A of Con1 with NS5B of JFH1, we observed more-efficient replication with the genotype 2a X tail, indicating that NS5B recognizes genotype-specific signals in this region. In contrast, a combination of the NS3 helicase with NS5A and NS5B was required to confer genotype specificity to the 5' NTR. These results present the first genetic evidence for an interaction between helicase, NS5A, and NS5B required for the initiation of RNA synthesis and provide a system for the specific analysis of HCV positive- and negative-strand syntheses.

* Corresponding author. Mailing address: University of Heidelberg, Department of Molecular Virology, Im Neuenheimer Feld 345, 69120 Heidelberg, Germany. Phone: 49 6221 566449. Fax: 49 6221 564570. E-mail: volker_lohmann{at}med.uni-heidelberg.de

{triangledown} Published ahead of print on 7 March 2007.

{dagger} These authors contributed equally to this study.

Journal of Virology, May 2007, p. 5270-5283, Vol. 81, No. 10
0022-538X/07/$08.00+0     doi:10.1128/JVI.00032-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Friebe, P., Bartenschlager, R. (2009). Role of RNA Structures in Genome Terminal Sequences of the Hepatitis C Virus for Replication and Assembly. J. Virol. 83: 11989-11995 [Abstract] [Full Text]  
  • Simister, P., Schmitt, M., Geitmann, M., Wicht, O., Danielson, U. H., Klein, R., Bressanelli, S., Lohmann, V. (2009). Structural and Functional Analysis of Hepatitis C Virus Strain JFH1 Polymerase. J. Virol. 83: 11926-11939 [Abstract] [Full Text]  
  • Gouttenoire, J., Montserret, R., Kennel, A., Penin, F., Moradpour, D. (2009). An Amphipathic {alpha}-Helix at the C Terminus of Hepatitis C Virus Nonstructural Protein 4B Mediates Membrane Association. J. Virol. 83: 11378-11384 [Abstract] [Full Text]  
  • Weinlich, S., Huttelmaier, S., Schierhorn, A., Behrens, S.-E., Ostareck-Lederer, A., Ostareck, D. H. (2009). IGF2BP1 enhances HCV IRES-mediated translation initiation via the 3'UTR. RNA 15: 1528-1542 [Abstract] [Full Text]  
  • Liu, Z., Yang, F., Robotham, J. M., Tang, H. (2009). Critical Role of Cyclophilin A and Its Prolyl-Peptidyl Isomerase Activity in the Structure and Function of the Hepatitis C Virus Replication Complex. J. Virol. 83: 6554-6565 [Abstract] [Full Text]  
  • Vassilaki, N., Friebe, P., Meuleman, P., Kallis, S., Kaul, A., Paranhos-Baccala, G., Leroux-Roels, G., Mavromara, P., Bartenschlager, R. (2008). Role of the Hepatitis C Virus Core+1 Open Reading Frame and Core cis-Acting RNA Elements in Viral RNA Translation and Replication. J. Virol. 82: 11503-11515 [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]  
  • Nikonov, A., Juronen, E., Ustav, M. (2008). Functional Characterization of Fingers Subdomain-specific Monoclonal Antibodies Inhibiting the Hepatitis C Virus RNA-dependent RNA Polymerase. J. Biol. Chem. 283: 24089-24102 [Abstract] [Full Text]  
  • Yang, F., Robotham, J. M., Nelson, H. B., Irsigler, A., Kenworthy, R., Tang, H. (2008). Cyclophilin A Is an Essential Cofactor for Hepatitis C Virus Infection and the Principal Mediator of Cyclosporine Resistance In Vitro. J. Virol. 82: 5269-5278 [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]  


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