Turnover rate of yeast PGK mRNA can be changed by specific alterations in its trailer structure

Biochimie. 1991 Jun;73(6):729-37. doi: 10.1016/0300-9084(91)90053-4.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Binding Sites
  • Chromosome Mapping
  • DNA Mutational Analysis
  • Molecular Sequence Data
  • Mutagenesis, Insertional
  • Phosphoglycerate Kinase / genetics*
  • Phosphoglycerate Kinase / metabolism
  • Poly G / genetics
  • Poly U / genetics
  • Protein Biosynthesis*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • Saccharomyces cerevisiae / genetics*
  • Transcription, Genetic

Substances

  • RNA, Messenger
  • Poly G
  • Poly U
  • Phosphoglycerate Kinase