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 Kohl, A.
Right arrow Articles by Elliott, R. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kohl, A.
Right arrow Articles by Elliott, R. M.

 Previous Article  |  Next Article 

Journal of Virology, June 2004, p. 5679-5685, Vol. 78, No. 11
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.11.5679-5685.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

A Bunyamwera Virus Minireplicon System in Mosquito Cells

Alain Kohl,1 Timothy J. Hart,1 Carol Noonan,1 Elizabeth Royall,2 Lisa O. Roberts,2 and Richard M. Elliott1*

Division of Virology, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G11 5JR, Scotland,1 School of Biomedical and Molecular Sciences, University of Surrey, Guildford GU2 7XH, England, United Kingdom2

Received 14 November 2003/ Accepted 28 January 2004

Artificial minigenomes are powerful tools for studying the replication and transcription of negative-strand RNA viruses. Bunyamwera virus (BUN; genus Orthobunyavirus, family Bunyaviridae) is an arbovirus that shows fundamental biological differences when replicating in mammalian versus mosquito cells. To study BUN RNA synthesis in mosquito cells, we developed a bacteriophage T7 RNA polymerase-based minireplicon system similar to that described previously for mammalian cells. An Aedes albopictus C6/36-derived mosquito cell line stably expressing T7 RNA polymerase was established. Viral proteins and artificial minigenomes (containing Renilla luciferase as a reporter) were transcribed and expressed in these cells from transfected T7 promoter-containing plasmids. Transcription of the minigenome required two viral proteins, the nucleocapsid protein N and the RNA-dependent RNA polymerase L, a situation similar to that in mammalian cells. However, unlike the situation in mammalian cells, the viral polymerase was not inhibited by the viral nonstructural protein NSs. We also report that promoter strength is different for vertebrate versus invertebrate cells. The development of this system opens the way for a detailed comparison of bunyavirus replication in cells of disparate phylogeny.

* Corresponding author. Mailing address: Institute of Virology, University of Glasgow, Church Street, Glasgow G11 5JR, Scotland, United Kingdom. Phone: 44 141 330 4024. Fax: 44 141 337 2236. E-mail: r.elliott{at}vir.gla.ac.uk.

Journal of Virology, June 2004, p. 5679-5685, Vol. 78, No. 11
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.11.5679-5685.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Eifan, S. A., Elliott, R. M. (2009). Mutational Analysis of the Bunyamwera Orthobunyavirus Nucleocapsid Protein Gene. J. Virol. 83: 11307-11317 [Abstract] [Full Text]  
  • Attarzadeh-Yazdi, G., Fragkoudis, R., Chi, Y., Siu, R. W. C., Ulper, L., Barry, G., Rodriguez-Andres, J., Nash, A. A., Bouloy, M., Merits, A., Fazakerley, J. K., Kohl, A. (2009). Cell-to-Cell Spread of the RNA Interference Response Suppresses Semliki Forest Virus (SFV) Infection of Mosquito Cell Cultures and Cannot Be Antagonized by SFV. J. Virol. 83: 5735-5748 [Abstract] [Full Text]  
  • Shi, X., Elliott, R. M. (2009). Generation and analysis of recombinant Bunyamwera orthobunyaviruses expressing V5 epitope-tagged L proteins. J. Gen. Virol. 90: 297-306 [Abstract] [Full Text]  
  • Shi, X., Kohl, A., Li, P., Elliott, R. M. (2007). Role of the Cytoplasmic Tail Domains of Bunyamwera Orthobunyavirus Glycoproteins Gn and Gc in Virus Assembly and Morphogenesis. J. Virol. 81: 10151-10160 [Abstract] [Full Text]  
  • Mir, M. A., Brown, B., Hjelle, B., Duran, W. A., Panganiban, A. T. (2006). Hantavirus N Protein Exhibits Genus-Specific Recognition of the Viral RNA Panhandle. J. Virol. 80: 11283-11292 [Abstract] [Full Text]  
  • Shi, X., Kohl, A., Leonard, V. H. J., Li, P., McLees, A., Elliott, R. M. (2006). Requirement of the N-Terminal Region of Orthobunyavirus Nonstructural Protein NSm for Virus Assembly and Morphogenesis.. J. Virol. 80: 8089-8099 [Abstract] [Full Text]  
  • Mir, M. A., Panganiban, A. T. (2006). Characterization of the RNA chaperone activity of hantavirus nucleocapsid protein.. J. Virol. 80: 6276-6285 [Abstract] [Full Text]  
  • Kohl, A., Lowen, A. C., Leonard, V. H. J., Elliott, R. M. (2006). Genetic elements regulating packaging of the Bunyamwera orthobunyavirus genome. J. Gen. Virol. 87: 177-187 [Abstract] [Full Text]  
  • Kohl, A., Dunn, E. F., Lowen, A. C., Elliott, R. M. (2004). Complementarity, sequence and structural elements within the 3' and 5' non-coding regions of the Bunyamwera orthobunyavirus S segment determine promoter strength. J. Gen. Virol. 85: 3269-3278 [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»