Proteomics

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Quantifying mistranslation from alanine tRNA anticodon variants in S. cerevisiae

ABSTRACT: Transfer RNAs (tRNAs) maintain translational fidelity through strict charging by their cognate aminoacyl-tRNA synthetase and codon:anticodon base pairing with the mRNA at the ribosome. Mistranslation occurs when an amino acid not specified by the genetic code is incorporated into a protein. Since alanyl-tRNA synthetase uniquely recognizes a G3:U70 base pair in alanine tRNAs and the anticodon plays no role in charging, alanine tRNA variants with anticodon mutations have the potential to mistranslate alanine. Our goal was to quantify mis-incorporation of alanine into proteins in Saccharomyces cerevisiae strains expressing one of 57 different alanine tRNA anticodon variants. Using mass spectrometry, we observed mistranslation for 45 of the variants when expressed on single-copy plasmids.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Matthew Berg  

LAB HEAD: Judit Villen

PROVIDER: PXD038242 | Pride | 2023-10-02

REPOSITORIES: Pride

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40042_e07254_MB013_tAlaMistrans_01_7A_20220312_2ul.fasta Fasta
40042_e07254_MB013_tAlaMistrans_01_7A_20220312_2ul.mzXML Mzxml
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40042_e07254_MB013_tAlaMistrans_01_7A_20220312_2ul.pepXML Pepxml
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Publications

Anticodon sequence determines the impact of mistranslating tRNA<sup>Ala</sup> variants.

Cozma Ecaterina E   Rao Megha M   Dusick Madison M   Genereaux Julie J   Rodriguez-Mias Ricard A RA   Villén Judit J   Brandl Christopher J CJ   Berg Matthew D MD  

RNA biology 20230101 1

Transfer RNAs (tRNAs) maintain translation fidelity through accurate charging by their cognate aminoacyl-tRNA synthetase and codon:anticodon base pairing with the mRNA at the ribosome. Mistranslation occurs when an amino acid not specified by the genetic message is incorporated into proteins and has applications in biotechnology, therapeutics and is relevant to disease. Since the alanyl-tRNA synthetase uniquely recognizes a G3:U70 base pair in tRNA<sup>Ala</sup> and the anticodon plays no role i  ...[more]

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