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Codon-Dependent Translational Accuracy Controls Protein Quality in Escherichia coli but not in Saccharomyces cerevisiae

Lyne Jossé, Connor D. D. Sampson, View ORCID ProfileMick F. Tuite, Kevin Howland, View ORCID ProfileTobias von der Haar
doi: https://doi.org/10.1101/200006
Lyne Jossé
Kent Fungal Group, School of Biosciences, University of Kent, Canterbury, CT2 7NJ, UK
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Connor D. D. Sampson
Kent Fungal Group, School of Biosciences, University of Kent, Canterbury, CT2 7NJ, UK
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Mick F. Tuite
Kent Fungal Group, School of Biosciences, University of Kent, Canterbury, CT2 7NJ, UK
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  • ORCID record for Mick F. Tuite
Kevin Howland
Kent Fungal Group, School of Biosciences, University of Kent, Canterbury, CT2 7NJ, UK
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  • For correspondence: T.von-der-Haar@kent.ac.uk K.Howland@kent.ac.uk
Tobias von der Haar
Kent Fungal Group, School of Biosciences, University of Kent, Canterbury, CT2 7NJ, UK
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  • ORCID record for Tobias von der Haar
  • For correspondence: T.von-der-Haar@kent.ac.uk K.Howland@kent.ac.uk
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Abstract

In order to generate a functional proteome, gene expression pathways must assemble proteins accurately according to the rules of the genetic code. General gene expression accuracy is known to be high, but errors nevertheless occur with measurable frequencies. Here we develop a mass-spectrometry (MS) based assay for the detection of a particular type of gene expression error, amino acid misincorporation. This assay allows assessing a much broader range of misincorporation events compared to current, very sensitive but also very specific enzyme reporter assays. Our assay uncovers a remarkably rich pool of error products for a model protein expressed in E. coli, which depend quantitatively on codon usage in the expression construct. This codon usage dependence can be explained in part as a function of the composition of the tRNA pool in this organism. We further show that codon-dependent differences in error levels correlate with measurable changes in specific protein activity. In contrast to E. coli, error levels are lower, and appear not to be codon usage dependent, when the same model protein is expressed in S. cerevisiae.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license.
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Posted December 21, 2017.
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Codon-Dependent Translational Accuracy Controls Protein Quality in Escherichia coli but not in Saccharomyces cerevisiae
Lyne Jossé, Connor D. D. Sampson, Mick F. Tuite, Kevin Howland, Tobias von der Haar
bioRxiv 200006; doi: https://doi.org/10.1101/200006
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Codon-Dependent Translational Accuracy Controls Protein Quality in Escherichia coli but not in Saccharomyces cerevisiae
Lyne Jossé, Connor D. D. Sampson, Mick F. Tuite, Kevin Howland, Tobias von der Haar
bioRxiv 200006; doi: https://doi.org/10.1101/200006

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