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 Tellinghuisen, T. L.
Right arrow Articles by Rice, C. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tellinghuisen, T. L.
Right arrow Articles by Rice, C. M.

 Previous Article  |  Next Article 

Journal of Virology, February 2008, p. 1073-1083, Vol. 82, No. 3
0022-538X/08/$08.00+0     doi:10.1128/JVI.00328-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Identification of Residues Required for RNA Replication in Domains II and III of the Hepatitis C Virus NS5A Protein{triangledown}

Timothy L. Tellinghuisen,1,2 Katie L. Foss,1 Jason C. Treadaway,1 and Charles M. Rice2*

The Scripps Research Institute, 5353 Parkside Drive, RF-2, Jupiter, Florida 33458,1 Laboratory of Virology and Infectious Disease, Center for the Study of Hepatitis C, The Rockefeller University, 1230 York Avenue, Box 64, New York, New York 100652

Received 14 February 2007/ Accepted 11 November 2007

The NS5A protein of hepatitis C virus (HCV) plays an important but undefined role in viral RNA replication. NS5A has been proposed to be a three-domain protein, and the crystal structure of the well-conserved amino-terminal domain I has been determined. The remaining two domains of NS5A, designated domains II and III, and their corresponding interdomain regions are poorly understood. We have conducted a detailed mutagenesis analysis of NS5A domains II and III using the genotype 1b HCV replicon system. The majority of the mutants containing 15 small (8- to 15-amino-acid) deletions analyzed were capable of efficient RNA replication. Only five deletion mutations yielded lethal phenotypes, and these were colinear, spanning a 56-amino-acid region within domain II. This region was further analyzed by combining triple and single alanine scanning mutagenesis to identify individual residues required for RNA replication. Based upon this analysis, 23 amino acids were identified that were found to be essential. In addition, two residues were identified that yielded a small colony phenotype while possessing only a moderate defect in RNA replication. These results indicate that the entire domain III region and large portions of domain II of the NS5A protein are not required for the function of NS5A in HCV RNA replication.

* Corresponding author. Mailing address: Laboratory of Virology and Infectious Disease, Center for the Study of Hepatitis C, The Rockefeller University, 1230 York Avenue, Box 64, New York, NY 10065. Phone: (212) 327-7046. Fax: (212) 327-7048. E-mail: ricec{at}rockefeller.edu

{triangledown} Published ahead of print on 21 November 2007.

Journal of Virology, February 2008, p. 1073-1083, Vol. 82, No. 3
0022-538X/08/$08.00+0     doi:10.1128/JVI.00328-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Xiao, M., Wang, Y., Zhu, Z., Yu, J., Wan, L., Chen, J. (2009). Influence of NS5A protein of classical swine fever virus (CSFV) on CSFV internal ribosome entry site-dependent translation. J. Gen. Virol. 90: 2923-2928 [Abstract] [Full Text]  
  • Hughes, M., Gretton, S., Shelton, H., Brown, D. D., McCormick, C. J., Angus, A. G. N., Patel, A. H., Griffin, S., Harris, M. (2009). A Conserved Proline between Domains II and III of Hepatitis C Virus NS5A Influences both RNA Replication and Virus Assembly. J. Virol. 83: 10788-10796 [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]  
  • Hughes, M., Griffin, S., Harris, M. (2009). Domain III of NS5A contributes to both RNA replication and assembly of hepatitis C virus particles. J. Gen. Virol. 90: 1329-1334 [Abstract] [Full Text]  
  • Weatherford, T., Chavez, D., Brasky, K. M., Lanford, R. E. (2009). The Marmoset Model of GB Virus B Infections: Adaptation to Host Phenotypic Variation. J. Virol. 83: 5806-5814 [Abstract] [Full Text]  
  • Hanoulle, X., Badillo, A., Wieruszeski, J.-M., Verdegem, D., Landrieu, I., Bartenschlager, R., Penin, F., Lippens, G. (2009). Hepatitis C Virus NS5A Protein Is a Substrate for the Peptidyl-prolyl cis/trans Isomerase Activity of Cyclophilins A and B. J. Biol. Chem. 284: 13589-13601 [Abstract] [Full Text]  
  • Love, R. A., Brodsky, O., Hickey, M. J., Wells, P. A., Cronin, C. N. (2009). Crystal Structure of a Novel Dimeric Form of NS5A Domain I Protein from Hepatitis C Virus. J. Virol. 83: 4395-4403 [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]  
  • Masaki, T., Suzuki, R., Murakami, K., Aizaki, H., Ishii, K., Murayama, A., Date, T., Matsuura, Y., Miyamura, T., Wakita, T., Suzuki, T. (2008). Interaction of Hepatitis C Virus Nonstructural Protein 5A with Core Protein Is Critical for the Production of Infectious Virus Particles. J. Virol. 82: 7964-7976 [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»