Mistranslation of human phosphoglycerate kinase in yeast in the presence of paromomycin.
Missense errors in the translation of mRNAs in Saccharomyces cerevisiae were screened by looking for charge heterogeneity of proteins on two-dimensional gels resulting from the substitution of charged and neutral amino acids. No such mistranslation was detected in wild-type yeast strains grown in the presence of the translational error-inducing antibiotic paromomycin. However, paromomycin-induced mistranslation of a heterologous mRNA, encoding human phosphoglycerate kinase expressed in yeast, was seen. We suggest that the combination of error-prone translation of a heterologous mRNA, and growth in the presence of paromomycin, leads to an accumulation of mistranslated proteins that can be detected by two-dimensional gel electrophoresis.
Grant
C M
CM
Biological Laboratory, University of Kent, Canterbury, UK.
Tuite
M F
MF
D019747:Aldose-Ketose Isomerases; D001345:Autoradiography; D003001:Cloning, Molecular / Q000187:drug effects*; D005656:Fungal Proteins / Q000096:biosynthesis; D005800:Genes, Fungal / Q000187:drug effects; D006801:Humans; D008715:Methionine / Q000378:metabolism; D010303:Paromomycin / Q000494:pharmacology*; D010735:Phosphoglycerate Kinase / Q000096:biosynthesis* / Q000235:genetics; D010957:Plasmids; D014176:Protein Biosynthesis / Q000187:drug effects*; D012333:RNA, Messenger / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D013462:Sulfur Radioisotopes
D005656:Fungal Proteins; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D013462:Sulfur Radioisotopes; D010303:Paromomycin; D008715:Methionine; D010735:Phosphoglycerate Kinase; D019747:Aldose-Ketose Isomerases; C490353:TRP1 protein, S cerevisiae