Starvation for His-tRNAHis in yeast causes translational arrest without a high level of misincorporation of glutamine at histidine codons.
The hts1.1 temperature-sensitive histidinyl-tRNA synthetase mutation enables Saccharomyces cerevisiae to be starved for His-tRNAHis by upshift to the non-permissive temperature of 38 degrees C. If yeast behaves similarly to bacterial and mammalian cells, this lack of His-tRNAHis should greatly enhance misreading at histidine codons (CAU/CAC) by Gln-tRNAGln, resulting in substitution of the neutral amino acid glutamine in place of histidine, a basic amino acid. Such misreading causes the isoelectric point (pI) of proteins to shift to lower values, and is readily detectable as "stuttering" on two-dimensional (2D) protein gels. By gel analysis of pulse-labelled proteins of hts1.1 yeast cells that were overexpressing phosphoglycerate kinase (PGK), our study sought to detect this specific translational error in PGK protein. It was not detected by this relatively sensitive technique, indicating that missense errors due to glutamine insertion at histidine codons do not occur in yeast at the readily-detectable level found in bacterial and mammalian cells.
Hirst
K
K
Department of Biochemistry and Molecular Biology, University College London, UK.
Piper
P W
PW
D003062:Codon; D005973:Glutamine / Q000235:genetics; D006639:Histidine / Q000235:genetics; D006643:Histidine-tRNA Ligase / Q000235:genetics; D009154:Mutation; D010735:Phosphoglycerate Kinase / Q000096:biosynthesis; D011132:Polyribosomes / Q000737:chemistry; D014176:Protein Biosynthesis / Q000502:physiology*; D012346:RNA, Transfer, Amino Acyl / Q000502:physiology; D012354:RNA, Transfer, His / Q000502:physiology*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013696:Temperature
D003062:Codon; D012346:RNA, Transfer, Amino Acyl; D012354:RNA, Transfer, His; D005973:Glutamine; D006639:Histidine; D010735:Phosphoglycerate Kinase; D006643:Histidine-tRNA Ligase