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Journal of Virology, November 2001, p. 10244-10249, Vol. 75, No. 21
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.21.10244-10249.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

The 5' Untranslated Region of Rhopalosiphum padi Virus Contains an Internal Ribosome Entry Site Which Functions Efficiently in Mammalian, Plant, and Insect Translation Systems

Kathryn E. Woolaway,1 Konstantinos Lazaridis,1,dagger Graham J. Belsham,2 Michael J. Carter,1 and Lisa O. Roberts1,*

School of Biomedical and Life Sciences, University of Surrey, Guildford GU2 7XH,1 and Institute for Animal Health, Pirbright, Woking, Surrey GU24 ONF,2 United Kingdom

Received 18 August 2000/Accepted 31 July 2001

Rhopalosiphum padi virus (RhPV) is one of several picorna-like viruses that infect insects; sequence analysis has revealed distinct differences between these agents and mammalian picornaviruses. RhPV has a single-stranded positive-sense RNA genome of about 10 kb; unlike the genomes of Picornaviridae, however, this genome contains two long open reading frames (ORFs). ORF1 encodes the virus nonstructural proteins, while the downstream ORF, ORF2, specifies the structural proteins. Both ORFs are preceded by long untranslated regions (UTRs). The intergenic UTR is known to contain an internal ribosome entry site (IRES) which directs non-AUG-initiated translation of ORF2. We have examined the 5' UTR of RhPV for IRES activity by translating synthetic dicistronic mRNAs containing this sequence in a variety of systems. We now report that the 5' UTR contains an element which directs internal initiation of protein synthesis from an AUG codon in mammalian, plant, and Drosophila in vitro translation systems. In contrast, the encephalomyocarditis virus IRES functions only in the mammalian system. The RhPV 5' IRES element has features in common with picornavirus IRES elements, in that no coding sequence is required for IRES function, but also with cellular IRES elements, as deletion analysis indicates that this IRES element does not have sharply defined boundaries.

* Corresponding author. Mailing address: School of Biomedical and Life Sciences, University of Surrey, Guildford GU2 7XH, United Kingdom. Phone: (44) 1483 686499. Fax: (44) 1483 300374. E-mail: l.roberts{at}surrey.ac.uk.

dagger Present address: School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, United Kingdom.

Journal of Virology, November 2001, p. 10244-10249, Vol. 75, No. 21
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.21.10244-10249.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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