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pubmed-article:1764519pubmed:abstractTextThe 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.lld:pubmed
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pubmed-article:1764519pubmed:dateRevised2006-11-15lld:pubmed
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pubmed-article:1764519pubmed:articleTitleTurnover rate of yeast PGK mRNA can be changed by specific alterations in its trailer structure.lld:pubmed
pubmed-article:1764519pubmed:affiliationBiochemisch Laboratorium, Vrije Universiteit, Amsterdam, The Netherlands.lld:pubmed
pubmed-article:1764519pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:1764519pubmed:publicationTypeResearch Support, Non-U.S. Gov'tlld:pubmed
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