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Effects of sequence elements in the potato virus X RNA 5' non-translated -leader on its translation enhancing activity

Department of Virology, A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow 119899, Russia

The 5' non-translated -leader sequence of potato virus X RNA consists of two regions: the sequence (41 nucleotides with no G) and the sequence (42 nucleotides upstream from AUG). The -leader has been shown to enhance strongly the expression of adjacent genes in chimeric mRNAs. This phenomenon has been postulated to be due to the unpaired conformation of the 5'-terminal 30 nucleotides and/or to the presence within the region of the CCACC pentanucleotide complementary to the 3'-terminal conserved structure of 18S rRNA. Different derivatives of -leader have been constructed for use in determining the contribution of separate elements of the sequence to translational enhancement. It was found that deletion of the sequence large fragment which was supposed to be unfolded did not reduce the -leader enhancement activity. Moreover, translational enhancement was greater for this derivative. Deletion of the sequence resulted in a considerable increase in activity of the -leader showing that the region was dispensable for translation. Disruption or ‘masking’ of CCACC led to inactivation of the -leader as a translational enhancer. Thus, we identified the CCACC pentanucleotide as the primary motif responsible for the translation enhancing ability of -leader.

Received 3 June 1993; accepted 28 July 1993.

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