The effect of insertions in the 5'- and 3'-untranslated regions (UTR) of the Saccharomyces cerevisiae mRNA encoding phosphoglycerate kinase (PGK) on the stability of the transcript in vivo was determined. None of the structural alterations in the 5'-UTR affected mRNA turnover significantly, despite the strong negative effect on translational efficiency of some of these alterations previously observed. We conclude that the structure of the 5'-UTR is not important for the relatively high affinity of PGK mRNA in yeast cells. Moreover, translation cannot be a major factor in determining the rate of turnover of this mRNA. Insertion of either a polyG or polyU, but not a polyA or polyC, tract into the 3'-UTR of PGK mRNA increased its half-life by a factor of about two. Introduction of a hairpin structure containing 18 G.C base pairs had only a slight stabilizing effect. We argue that the stabilization by the structural changes in the 3'-UTR is due to altered folding of the mutant mRNA which retards a rate-limiting endonucleolytic cleavage step in the normal turnover pathway of PGK mRNA. The stabilizing effect of local structural alterations in the 3'-UTR opens the possibility for further increasing the product yield of a (heterologous) gene cloned in yeast cells.

• PMID: 1764519
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Journal Article | Research Support, Non-U.S. Gov't

Authors

Vreken P, van der Veen R, de Regt VC, de Maat AL, Planta RJ, Raué HA

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