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Journal of Virology, June 2002, p. 5678-5691, Vol. 76, No. 11
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.11.5678-5691.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Eukaryotic Elongation Factor 1A Interacts with the Upstream Pseudoknot Domain in the 3' Untranslated Region of Tobacco Mosaic Virus RNA

Vladimir V. Zeenko,1,{dagger} Lyubov A. Ryabova,1,{ddagger} Alexander S. Spirin,2 Helen M. Rothnie,1 Daniel Hess,1 Karen S. Browning,3 and Thomas Hohn1*

Friedrich Miescher Institute, CH-4002 Basel, Switzerland,1 Institute of Protein Research, Russian Academy of Science, Pushchino, Russia,2 Department of Chemistry and Biochemistry, University of Texas, Austin, Texas 787123

Received 11 September 2001/ Accepted 1 March 2002

The genomic RNA of tobacco mosaic virus (TMV), like that of other positive-strand RNA viruses, acts as a template for both translation and replication. The highly structured 3' untranslated region (UTR) of TMV RNAs plays an important role in both processes; it is not polyadenylated but ends with a tRNA-like structure (TLS) preceded by a conserved upstream pseudoknot domain (UPD). The TLS of tobamoviral RNAs can be specifically aminoacylated and, in this state, can interact with eukaryotic elongation factor 1A (eEF1A)/GTP with high affinity. Using a UV cross-linking assay, we detected another specific binding site for eEF1A/GTP, within the UPDs of TMV and crucifer-infecting tobamovirus (crTMV), that does not require aminoacylation. A mutational analysis revealed that UPD pseudoknot conformation and some conserved primary sequence elements are required for this interaction. Its possible role in the regulation of tobamovirus gene expression and replication is discussed.


* Corresponding author. Mailing address: Friedrich Miescher Institute, P.O. Box 2543, CH-4002 Basel, Switzerland. Phone: (41) 61 697 72 66. Fax: (41) 61 697 39 76. E-mail: thomas.hohn{at}fmi.ch.

{dagger} Present address: Department of Biochemistry, University of California, Riverside, CA 92521.

{ddagger} Present address: Protéus S.A., 30000 Nîmes, France.


Journal of Virology, June 2002, p. 5678-5691, Vol. 76, No. 11
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.11.5678-5691.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.




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