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The influence of mutations at ATG triplets of the open reading frame SUP35 on viability of the yeast Saccharomyces cerevisiae

Ryabinkova, N. A., Borchsenius, A. S., Inge-Vechtomov, S. G.
Russian journal of genetics 2009 v.45 no.2 pp. 155-160
Saccharomyces cerevisiae, binding sites, carbon, codons, environmental factors, galactose, gene expression, glucose, histidine, messenger RNA, mutants, open reading frames, peptides, point mutation, starvation, transcription factors, translation (genetics), viability, yeasts
The open reading frame SUP35 encoding the translation termination eRF3 factor vital to life contains three ATG codons (ATG₁, ATG₁₂₄, and ATG₂₅₄). Previously, other authors detected two SUP35 transcripts: a major one that corresponds to the full-length open reading frame and a minor transcript that corresponds to the 3' terminal site of SUP35 starting at the third ATG codon (ATG₂₅₄). In this work, mutations at triplets ATG₁, ATG₁₂₄, and ATG₂₅₄ were obtained as well as double mutations, which combine the point mutation in one of three ATG triplets and a deletion at the site for binding with the transcription factor Abf1 within the SUP35 (sup35-ΔAbf1) promoter. The influence of these mutations on the yeast viability was analyzed. Mutations at triplets ATG₁₂₄ and ATG₂₅₄ did not affect yeast viability in their own right or in the background of deletion sup35-ΔAbf1. Mutation sup35-AGG ₁ (ATG₁ [rightward arrow] AGG) causes the lethal effect in cells grown on media containing glucose as the sole source of carbon. The replacement of glucose by galactose, or histidine starvation, partially restore the viability of sup35-AGG ₁ mutants, but not that of double mutants sup35-ΔAbf1,AGG ₁ . The restoration of sup35-AGG ₁ mutant viability under these conditions can be explained by either the appearance (or enhancement) of the production of short peptides synthesized on the mRNA triplets SUP35 AUG₁₂₄ and AUG₂₅₄, or by the enhanced production of the full-length SUP35 transcript coupled with translation initiation from the noncanonical AGG₁ codon. These data confirm that the expression of gene SUP35 at the transcription and(or) translation level is regulated by environmental conditions.