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Journal of Virology, April 2004, p. 4048-4053, Vol. 78, No. 8
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.8.4048-4053.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Similarities and Differences between the Subgenomic and Minus-Strand Promoters of an RNA Plant Virus

René C. L. Olsthoorn,{dagger}* P. C. Joost Haasnoot,{ddagger} and John F. Bol

Institute of Biology, Gorlaeus Laboratories, Leiden University, 2300 RA Leiden, The Netherlands

Received 15 September 2003/ Accepted 18 December 2003

Promoter regions required for minus-strand and subgenomic RNA synthesis have been mapped for several plus-strand RNA viruses. In general, the two types of promoters do not share structural features even though they are recognized by the same viral polymerase. The minus-strand promoter of Alfalfa mosaic virus (AMV), a plant virus of the family Bromoviridae, consists of a triloop hairpin (hpE) which is attached to a 3' tRNA-like structure (TLS). In contrast, the AMV subgenomic promoter consists of a single triloop hairpin that bears no sequence homology with hpE. Here we show that hpE, when detached from its TLS, can function as a subgenomic promoter in vitro and can replace the authentic subgenomic promoter in the live virus. Thus, the AMV subgenomic and minus-strand promoters are basically the same, but the minus-strand promoter is linked to a 3' TLS to force the polymerase to initiate at the very 3'end.

* Corresponding author. Mailing address: Leiden Institute of Chemistry, Gorlaeus Laboratories, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands. Phone: 31-71-5274586. Fax: 31-71-5274340. E-mail: olsthoor{at}chem.leidenuniv.nl.

{dagger} Present address: Leiden Institute of Chemistry, Gorlaeus Laboratories, Leiden University, 2300 RA Leiden, The Netherlands.

{ddagger} Present address: Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands.

Journal of Virology, April 2004, p. 4048-4053, Vol. 78, No. 8
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.8.4048-4053.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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