The genome of plum pox virus contains a single open reading frame that is translated into a large polyprotein. Although the open reading frame starts at nucleotide 36 (36AUG), it is translated from the second, 147AUG, which is in a more favourable context for translation initiation. We have carried out in vitro translation and transient expression analysis in protoplasts of a nested set of substitution and deletion mutants, and the results show that no internal structure in the 5' noncoding region of plum pox virus is necessary for efficient translation initiation. On the other hand, when the cryptic 36AUG was placed in a favourable context, it turned into an efficient initiation codon in vitro. Furthermore, AUGs that were placed in a favourable context, initiating short intraleader open reading frames, repressed translation initiation from the 147AUG in vitro and in vivo. These results point to leaky scanning as the mechanism of translation initiation of plum pox virus RNA. Nevertheless, it is a peculiar leaky scanning where the initiation of translation does not require a cap structure at the 5' end. This fact is congruent with the experimentally predicted absence of a stable secondary structure at the 5' noncoding region.


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