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 van den Born, E.
Right arrow Articles by Snijder, E. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by van den Born, E.
Right arrow Articles by Snijder, E. J.

 Previous Article  |  Next Article 

Journal of Virology, May 2005, p. 6312-6324, Vol. 79, No. 10
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.10.6312-6324.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Discontinuous Subgenomic RNA Synthesis in Arteriviruses Is Guided by an RNA Hairpin Structure Located in the Genomic Leader Region

Erwin van den Born,1 Clara C. Posthuma,1 Alexander P. Gultyaev,2 and Eric J. Snijder1*

Molecular Virology Laboratory, Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden, The Netherlands,1 Section Theoretical Biology, Leiden Institute of Biology, Leiden University, Leiden, The Netherlands2

Received 17 September 2004/ Accepted 28 December 2004

Nidoviruses produce an extensive 3'-coterminal nested set of subgenomic (sg) mRNAs, which are used to express structural proteins and sometimes accessory proteins. In arteriviruses and coronaviruses, these mRNAs contain a common 5' leader sequence, derived from the genomic 5' end. The joining of the leader sequence to different segments derived from the 3'-proximal part of the genome (mRNA bodies) presumably involves a unique mechanism of discontinuous minus-strand RNA synthesis in which base pairing between sense and antisense transcription-regulating sequences (TRSs) plays an essential role. The leader TRS is present in the loop of a hairpin structure that functions in sg mRNA synthesis. In this study, the minimal sequences in the 5'-proximal region of the Equine arteritis virus genome that are required for sg RNA synthesis were delimited through mutagenesis. A full-length cDNA clone was engineered in which this domain was duplicated, allowing us to make mutations and monitor their effects on sg RNA synthesis without seriously affecting genome replication and translation. The leader TRS present in the duplicated sequence was used and yielded novel sg mRNAs with significantly extended leaders. Our combined findings suggest that the leader TRS hairpin (LTH) and its immediate flanking sequences are essential for efficient sg RNA synthesis and form an independent functional entity that could be moved 300 nucleotides downstream of its original position in the genome. We hypothesize that a conformational switch in the LTH region regulates the role of the 5'-proximal region of the arterivirus genome in subgenomic RNA synthesis.

* Corresponding author. Mailing address: Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, LUMC P4-26, PO Box 9600, 2300 RC Leiden, The Netherlands. Phone: 31 71 5261657. Fax: 31 71 5266761. E-mail: e.j.snijder{at}lumc.nl.

Journal of Virology, May 2005, p. 6312-6324, Vol. 79, No. 10
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.10.6312-6324.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Tijms, M. A., Nedialkova, D. D., Zevenhoven-Dobbe, J. C., Gorbalenya, A. E., Snijder, E. J. (2007). Arterivirus Subgenomic mRNA Synthesis and Virion Biogenesis Depend on the Multifunctional nsp1 Autoprotease. J. Virol. 81: 10496-10505 [Abstract] [Full Text]  
  • Beerens, N., Snijder, E. J. (2007). An RNA Pseudoknot in the 3' End of the Arterivirus Genome Has a Critical Role in Regulating Viral RNA Synthesis. J. Virol. 81: 9426-9436 [Abstract] [Full Text]  
  • Liu, P., Li, L., Millership, J. J., Kang, H., Leibowitz, J. L., Giedroc, D. P. (2007). A U-turn motif-containing stem-loop in the coronavirus 5' untranslated region plays a functional role in replication. RNA 13: 763-780 [Abstract] [Full Text]  
  • van den Born, E., Posthuma, C. C., Knoops, K., Snijder, E. J. (2007). An infectious recombinant equine arteritis virus expressing green fluorescent protein from its replicase gene. J. Gen. Virol. 88: 1196-1205 [Abstract] [Full Text]  
  • Wierzchoslawski, R., Urbanowicz, A., Dzianott, A., Figlerowicz, M., Bujarski, J. J. (2006). Characterization of a Novel 5' Subgenomic RNA3a Derived from RNA3 of Brome Mosaic Bromovirus. J. Virol. 80: 12357-12366 [Abstract] [Full Text]  
  • Pasternak, A. O., Spaan, W. J. M., Snijder, E. J. (2006). Nidovirus transcription: how to make sense...?. J. Gen. Virol. 87: 1403-1421 [Abstract] [Full Text]  
  • van Ooij, M. J. M., Polacek, C., Glaudemans, D. H. R. F., Kuijpers, J., van Kuppeveld, F. J. M., Andino, R., Agol, V. I., Melchers, W. J. G. (2006). Polyadenylation of genomic RNA and initiation of antigenomic RNA in a positive-strand RNA virus are controlled by the same cis-element. Nucleic Acids Res 34: 2953-2965 [Abstract] [Full Text]  
  • Posthuma, C. C., Nedialkova, D. D., Zevenhoven-Dobbe, J. C., Blokhuis, J. H., Gorbalenya, A. E., Snijder, E. J. (2006). Site-Directed Mutagenesis of the Nidovirus Replicative Endoribonuclease NendoU Exerts Pleiotropic Effects on the Arterivirus Life Cycle. J. Virol. 80: 1653-1661 [Abstract] [Full Text]  
  • van den Born, E., Stein, D. A., Iversen, P. L., Snijder, E. J. (2005). Antiviral activity of morpholino oligomers designed to block various aspects of Equine arteritis virus amplification in cell culture. J. Gen. Virol. 86: 3081-3090 [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»