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 Ranjith-Kumar, C. T.
Right arrow Articles by Kao, C. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ranjith-Kumar, C. T.
Right arrow Articles by Kao, C. C.

 Previous Article  |  Next Article 

Journal of Virology, December 2002, p. 12526-12536, Vol. 76, No. 24
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.24.12526-12536.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Requirements for De Novo Initiation of RNA Synthesis by Recombinant Flaviviral RNA-Dependent RNA Polymerases

C. T. Ranjith-Kumar,1 Les Gutshall,2 Min-Ju Kim,1 Robert T. Sarisky,2 and C. Cheng Kao1*

Department of Biology, Indiana University, Bloomington, Indiana 47405,1 Department of Virology, The Metabolic and Antiviral Diseases Center of Excellence for Drug Discovery, GlaxoSmithKline Pharmaceuticals, Collegeville, Pennsylvania 194262

Received 7 May 2002/ Accepted 10 September 2002

RNA-dependent RNA polymerases (RdRps) that initiate RNA synthesis by a de novo mechanism should specifically recognize the template initiation nucleotide, T1, and the substrate initiation nucleotide, the NTPi. The RdRps from hepatitis C virus (HCV), bovine viral diarrhea virus (BVDV), and GB virus-B all can initiate RNA synthesis by a de novo mechanism. We used RNAs and GTP analogs, respectively, to examine the use of the T1 nucleotide and the initiation nucleotide (NTPi) during de novo initiation of RNA synthesis. The effects of the metal ions Mg2+ and Mn2+ on initiation were also analyzed. All three viral RdRps require correct base pairing between the T1 and NTPi for efficient RNA synthesis. However, each RdRp had some distinct tolerances for modifications in the T1 and NTPi. For example, the HCV RdRp preferred an NTPi lacking one or more phosphates regardless of whether Mn2+ was present or absent, while the BVDV RdRp efficiently used GDP and GMP for initiation of RNA synthesis only in the presence of Mn2+. These and other results indicate that although the three RdRps share a common mechanism of de novo initiation, each has distinct preferences.

* Corresponding author. Mailing address: Department of Biology, Indiana University, 1001 E. Third St., Bloomington, IN 47405. Phone: (812) 855-7959. Fax: (812) 855-6705. E-mail: ckao{at}bio.indiana.edu.

Journal of Virology, December 2002, p. 12526-12536, Vol. 76, No. 24
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.24.12526-12536.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • 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]  
  • Nedialkova, D. D., Ulferts, R., van den Born, E., Lauber, C., Gorbalenya, A. E., Ziebuhr, J., Snijder, E. J. (2009). Biochemical Characterization of Arterivirus Nonstructural Protein 11 Reveals the Nidovirus-Wide Conservation of a Replicative Endoribonuclease. J. Virol. 83: 5671-5682 [Abstract] [Full Text]  
  • Poranen, M. M., Salgado, P. S., Koivunen, M. R. L., Wright, S., Bamford, D. H., Stuart, D. I., Grimes, J. M. (2008). Structural explanation for the role of Mn2+ in the activity of {phi}6 RNA-dependent RNA polymerase. Nucleic Acids Res 36: 6633-6644 [Abstract] [Full Text]  
  • Steil, B. P., Barton, D. J. (2008). Poliovirus cis-Acting Replication Element-Dependent VPg Uridylylation Lowers the Km of the Initiating Nucleoside Triphosphate for Viral RNA Replication. J. Virol. 82: 9400-9408 [Abstract] [Full Text]  
  • Poranen, M. M., Koivunen, M. R. L., Bamford, D. H. (2008). Nontemplated Terminal Nucleotidyltransferase Activity of Double-Stranded RNA Bacteriophage {phi}6 RNA-Dependent RNA Polymerase. J. Virol. 82: 9254-9264 [Abstract] [Full Text]  
  • Chinnaswamy, S., Yarbrough, I., Palaninathan, S., Kumar, C. T. R., Vijayaraghavan, V., Demeler, B., Lemon, S. M., Sacchettini, J. C., Kao, C. C. (2008). A Locking Mechanism Regulates RNA Synthesis and Host Protein Interaction by the Hepatitis C Virus Polymerase. J. Biol. Chem. 283: 20535-20546 [Abstract] [Full Text]  
  • Deval, J., D'Abramo, C. M., Zhao, Z., McCormick, S., Coutsinos, D., Hess, S., Kvaratskhelia, M., Gotte, M. (2007). High Resolution Footprinting of the Hepatitis C Virus Polymerase NS5B in Complex with RNA. J. Biol. Chem. 282: 16907-16916 [Abstract] [Full Text]  
  • Malet, H., Egloff, M.-P., Selisko, B., Butcher, R. E., Wright, P. J., Roberts, M., Gruez, A., Sulzenbacher, G., Vonrhein, C., Bricogne, G., Mackenzie, J. M., Khromykh, A. A., Davidson, A. D., Canard, B. (2007). Crystal Structure of the RNA Polymerase Domain of the West Nile Virus Non-structural Protein 5. J. Biol. Chem. 282: 10678-10689 [Abstract] [Full Text]  
  • Howe, A. Y. M., Cheng, H., Thompson, I., Chunduru, S. K., Herrmann, S., O'Connell, J., Agarwal, A., Chopra, R., Del Vecchio, A. M. (2006). Molecular Mechanism of a Thumb Domain Hepatitis C Virus Nonnucleoside RNA-Dependent RNA Polymerase Inhibitor. Antimicrob. Agents Chemother. 50: 4103-4113 [Abstract] [Full Text]  
  • D'Abramo, C. M., Deval, J., Cameron, C. E., Cellai, L., Gotte, M. (2006). Control of Template Positioning during de Novo Initiation of RNA Synthesis by the Bovine Viral Diarrhea Virus NS5B Polymerase. J. Biol. Chem. 281: 24991-24998 [Abstract] [Full Text]  
  • RANJITH-KUMAR, C.T., KAO, C.C. (2006). Recombinant viral RdRps can initiate RNA synthesis from circular templates. RNA 12: 303-312 [Abstract] [Full Text]  
  • Dutartre, H., Boretto, J., Guillemot, J. C., Canard, B. (2005). A Relaxed Discrimination of 2'-O-Methyl-GTP Relative to GTP between de Novo and Elongative RNA Synthesis by the Hepatitis C RNA-dependent RNA Polymerase NS5B. J. Biol. Chem. 280: 6359-6368 [Abstract] [Full Text]  
  • Laurila, M. R. L., Salgado, P. S., Stuart, D. I., Grimes, J. M., Bamford, D. H. (2005). Back-priming mode of {phi}6 RNA-dependent RNA polymerase. J. Gen. Virol. 86: 521-526 [Abstract] [Full Text]  
  • Ranjith-Kumar, C. T., Sarisky, R. T., Gutshall, L., Thomson, M., Kao, C. C. (2004). De Novo Initiation Pocket Mutations Have Multiple Effects on Hepatitis C Virus RNA-Dependent RNA Polymerase Activities. J. Virol. 78: 12207-12217 [Abstract] [Full Text]  
  • Wierzchoslawski, R., Dzianott, A., Bujarski, J. (2004). Dissecting the Requirement for Subgenomic Promoter Sequences by RNA Recombination of Brome Mosaic Virus In Vivo: Evidence for Functional Separation of Transcription and Recombination. J. Virol. 78: 8552-8564 [Abstract] [Full Text]  
  • Kalinina, O., Norder, H., Magnius, L. O. (2004). Full-length open reading frame of a recombinant hepatitis C virus strain from St Petersburg: proposed mechanism for its formation. J. Gen. Virol. 85: 1853-1857 [Abstract] [Full Text]  
  • van Dijk, A. A., Makeyev, E. V., Bamford, D. H. (2004). Initiation of viral RNA-dependent RNA polymerization. J. Gen. Virol. 85: 1077-1093 [Abstract] [Full Text]  
  • Lee, K. J., Choi, J., Ou, J.-h., Lai, M. M. C. (2004). The C-Terminal Transmembrane Domain of Hepatitis C Virus (HCV) RNA Polymerase Is Essential for HCV Replication In Vivo. J. Virol. 78: 3797-3802 [Abstract] [Full Text]  
  • Choi, K. H., Groarke, J. M., Young, D. C., Kuhn, R. J., Smith, J. L., Pevear, D. C., Rossmann, M. G. (2004). The structure of the RNA-dependent RNA polymerase from bovine viral diarrhea virus establishes the role of GTP in de novo initiation. Proc. Natl. Acad. Sci. USA 101: 4425-4430 [Abstract] [Full Text]  
  • Gu, B., Johnston, V. K., Gutshall, L. L., Nguyen, T. T., Gontarek, R. R., Darcy, M. G., Tedesco, R., Dhanak, D., Duffy, K. J., Kao, C. C., Sarisky, R. T. (2003). Arresting Initiation of Hepatitis C Virus RNA Synthesis Using Heterocyclic Derivatives. J. Biol. Chem. 278: 16602-16607 [Abstract] [Full Text]  
  • ZHANG, X., KIM, C.-H., SIVAKUMARAN, K., KAO, C. (2003). Stable RNA structures can repress RNA synthesis in vitro by the brome mosaic virus replicase. RNA 9: 555-565 [Abstract] [Full Text]  
  • Ranjith-Kumar, C. T., Kim, Y.-C., Gutshall, L., Silverman, C., Khandekar, S., Sarisky, R. T., Kao, C. C. (2002). Mechanism of De Novo Initiation by the Hepatitis C Virus RNA-Dependent RNA Polymerase: Role of Divalent Metals. J. Virol. 76: 12513-12525 [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»