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Volume 270, Number 22, Issue of June 2, pp. 13446-13452, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.

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Shanping Wang , W. Allen Miller

An infectious, in vitro transcript from a full-length cDNA clone of the barley yellow dwarf virus (PAV serotype) genome translated efficiently in a wheat germ translation extract. Deletions in a region that we call the 3` translational enhancer, located between bases 4,513 and 5,009 in the 5,677-base genome, reduced translation of the 5`-proximal open reading frames from uncapped RNA by at least 30-fold. Deletions elsewhere in all but the 5` end of the genome had no effect on translation. Presence of a mG(5`)ppp(5`)G cap on the 5` end fully restored translational efficiency of transcripts lacking the 3` translational enhancer. The translation enhancer reduced inhibition of translation by free cap analog, did not affect RNA stability, and did not function in reticulocyte lysates. When placed in the 3`-untranslated region of uncapped mRNA encoding the -glucuronidase gene, the translation enhancer stimulated translation more than 80-fold, in the presence of the viral, but not a plasmid-derived, 5` leader. A polyadenylate tail could not substitute for the 3` translation enhancer. These observations provide an extreme example, in terms of distance from the 5` end and level of stimulation, of an mRNA in which a sequence near the 3` end stimulates translation.

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