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Author Notes:

E-mail: jpresteg@ccrc.uga.edu

We thank Christine Dunham of the Emory School of Medicine for her thoughtful critique of the initial version of this manuscript.

Competing Interests: The authors have declared that no competing interests exist

Subjects:

Research Funding:

This work was partially funded by a grant from the National Institute of General Medical Sciences to JHP, GM061268. FB was funded by EU Programme iNEXT.

Work supervised by HS was funded by the state of Hesse, Germany.

Keywords:

  • Science & Technology
  • Multidisciplinary Sciences
  • Science & Technology - Other Topics
  • RARE ARGININE CODON
  • ESCHERICHIA-COLI
  • MESSENGER-RNA
  • ACID MISINCORPORATION
  • PROTEIN EXPRESSION
  • RIBOSOME
  • CONTEXT
  • LEVEL
  • GENE
  • DYNAMICS

Mistakes in translation: Reflections on mechanism

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Journal Title:

PLoS ONE

Volume:

Volume 12, Number 6

Publisher:

, Pages e0180566-e0180566

Type of Work:

Article | Final Publisher PDF

Abstract:

Mistakes in translation of messenger RNA into protein are clearly a detriment to the recombinant production of pure proteins for biophysical study or the biopharmaceutical market. However, they may also provide insight into mechanistic details of the translation process. Mistakes often involve the substitution of an amino acid having an abundant codon for one having a rare codon, differing by substitution of a G base by an A base, as in the case of substitution of a lysine (AAA) for arginine (AGA). In these cases one expects the substitution frequency to depend on the relative abundances of the respective tRNAs, and thus, one might expect frequencies to be similar for all sites having the same rare codon. Here we demonstrate that, for the ADP-ribosylation factor from yeast expressed in E. coli, lysine for arginine substitutions frequencies are not the same at the 9 sites containing a rare arginine codon; mis-incorporation frequencies instead vary from less than 1 to 16%. We suggest that the context in which the codons occur (clustering of rare sites) may be responsible for the variation. The method employed to determine the frequency of mis-incorporation involves a novel mass spectrometric analysis of the products from the parallel expression of wild type and codon-optimized genes in 15N and 14N enriched media, respectively. The high sensitivity and low material requirements of the method make this a promising technology for the collection of data relevant to other mis-incorporations. The additional data could be of value in refining models for the ribosomal translation elongation process.

Copyright information:

© 2017 Liu et al

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/).
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