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Journal of Virology, December 2001, p. 12141-12152, Vol. 75, No. 24
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.24.12141-12152.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Cap-Independent Translation Conferred by the 5' Leader of Tobacco Etch Virus Is Eukaryotic Initiation Factor 4G Dependent

Daniel R. Gallie*

Department of Biochemistry, University of California, Riverside, California 92521-0129

Received 9 May 2001/Accepted 18 September 2001

The 5' leader of tobacco etch virus (TEV) genomic RNA directs efficient translation from the naturally uncapped viral mRNA. Two distinct regions within the TEV 143-nucleotide leader confer cap-independent translation in vivo even when present in the intercistronic region of a discistronic mRNA, indicating that the TEV leader contains an internal ribosome entry site (IRES). In this study, the requirements for TEV IRES activity were investigated. The TEV IRES enhanced translation of monocistronic or dicistronic mRNAs in vitro under competitive conditions, i.e., at high RNA concentration or in lysate partially depleted of eukaryotic initiation factor 4F (eIF4F) and eIFiso4F, the two cap binding complexes in plants. The translational advantage conferred by the TEV IRES under these conditions was lost when the lysate reduced in eIF4F and eIFiso4F was supplemented with eIF4F (or, to a lesser extent, eIFiso4F) but not when supplemented with eIF4E, eIFiso4E, eIF4A, or eIF4B. eIF4G, the large subunit of eIF4F, was responsible for the competitive advantage conferred by the TEV IRES. TEV IRES activity was enhanced moderately by the poly(A)-binding protein. These observations suggest that the TEV IRES directs cap-independent translation through a mechanism that involves eIF4G specifically.

* Mailing address: Department of Biochemistry, University of California, Riverside, CA 92521-0129. Phone: (909) 787-7298. Fax: (909) 787-3590. E-mail: drgallie{at}citrus.ucr.edu.

Journal of Virology, December 2001, p. 12141-12152, Vol. 75, No. 24
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.24.12141-12152.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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