Mutations in elongation factor EF-1 alpha affect the frequency of frameshifting and amino acid misincorporation in Saccharomyces cerevisiae.
A mutational analysis of the eukaryotic elongation factor EF-1 alpha indicates that this protein functions to limit the frequency of errors during genetic code translation. We found that both amino acid misincorporation and reading frame errors are controlled by EF-1 alpha. In order to examine the function of this protein, the TEF2 gene, which encodes EF-1 alpha in Saccharomyces cerevisiae, was mutagenized in vitro with hydroxylamine. Sixteen independent TEF2 alleles were isolated by their ability to suppress frameshift mutations. DNA sequence analysis identified eight different sites in the EF-1 alpha protein that elevate the frequency of mistranslation when mutated. These sites are located in two different regions of the protein. Amino acid substitutions located in or near the GTP-binding and hydrolysis domain of the protein cause suppression of frameshift and nonsense mutations. These mutations may effect mistranslation by altering the binding or hydrolysis of GTP. Amino acid substitutions located adjacent to a putative aminoacyl-tRNA binding region also suppress frameshift and nonsense mutations. These mutations may alter the binding of aminoacyl-tRNA by EF-1 alpha. The identification of frameshift and nonsense suppressor mutations in EF-1 alpha indicates a role for this protein in limiting amino acid misincorporation and reading frame errors. We suggest that these types of errors are controlled by a common mechanism or closely related mechanisms.
Sandbaken
M G
MG
Laboratory of Molecular Biology, University of Wisconsin, Madison 53706.
Culbertson
M R
MR
D000596:Amino Acids / Q000235:genetics*; D003062:Codon; D004252:DNA Mutational Analysis; D004271:DNA, Fungal; D005800:Genes, Fungal; D019811:Hydroxylamine; D006898:Hydroxylamines; D010303:Paromomycin / Q000494:pharmacology; D010441:Peptide Chain Elongation, Translational; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000235:genetics*; D014176:Protein Biosynthesis; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013489:Suppression, Genetic
D000596:Amino Acids; D003062:Codon; D004271:DNA, Fungal; D006898:Hydroxylamines; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D019811:Hydroxylamine; D010303:Paromomycin