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. 2015 Jan;21(1):124–134. doi: 10.1261/rna.047282.114

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© 2014 Roque et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 4. — Translational readthrough and polysome profiles of the mutant strains. (A) Translational readthrough levels determined with the dual luciferase reporter in strains SKY1405/1488, SKY1486, SKY1407, SKY1489, SKY1406, SKY1404 expressing the tRNA suppressor of UAA stop codon. Readthrough was quantified as described in the main text and in the Materials and Methods. SEM is indicated. The results were similar at 25°C or 30°C. (B) Strains used in A were streaked 3 d at 30°C on synthetic medium in the presence (+Ade) or the absence (−Ade) of adenine to assay the suppression of ade2–1 nonsense mutation. The result was identical at 25°C. (C) Extracts from strains SKY1264, SKY1266, BSY1537, SKY1265, SKY1263, and SKY1247 were separated on sucrose gradient by ultracentrifugation and absorbance at 254 nm was measured. The arrowheads indicate the position of stalled preinitiation complexes (halfmers). The same results were obtained with strains containing the same deleted or wild-type versions of the genes in parental strains or isolated from different tetrads.