Oxidation of cellular amino acid pools leads to cytotoxic mistranslation of the genetic code

Thank you for sending your work entitled 'Oxidation of cellular amino acid pools leads to cytotoxic mistranslation of the genetic code' for consideration at eLife. Your article has been favorably evaluated by a Senior editor, a Reviewing editor, and three reviewers.

The Reviewing editor has assembled the following comments to help you prepare a revised submission. All reviewers are enthusiastic about your study. One reviewer states: “Overall, the experiments were well described and the results clear. Most importantly, the conceptual advance is significant and likely to be of interest to the broad readers of eLife.” However, the reviewers feel two points need to be addressed before publication.

1) All reviewers felt the most interesting results in the manuscript focused on the E. coli story and found the yeast story distracting. Thus the authors should simply refer to the differences with yeast throughout, vaguely, and instead focus instead on the very tidy E. coli story so that the reader can take away this simple and elegant message.

2) The study would be strengthened if the authors could show that some m-Tyr indeed is incorporated to during oxidative stress since this is the main argument of the study. The reviewers understand that the low m-Tyr levels may present a challenge, but hope the experiment is do-able since data on m-Tyr incorporation into the proteome when cells are grown in m-Tyr are already provided.

We read your and the reviewers’ comments with interest and fully agree that the manuscript would greatly benefit from the revisions suggested. Consequently, a number of changes have been made which are detailed below, with reference to the specific comments of yourself and each of the referees.

1) All reviewers felt the most interesting results in the manuscript focused on the E. coli story and found the yeast story distracting. Thus the authors should simply refer to the differences with yeast throughout, vaguely, and instead focus instead on the very tidy E. coli story so that the reader can take away this simple and elegant message.

Yeast data on the UPR and on long-term survival have now been removed, and the only remaining results are those that refer to differences with E. coli.

2) The study would be strengthened if the authors could show that some m-Tyr indeed is incorporated to during oxidative stress since this is the main argument of the study. The reviewers understand that the low m-Tyr levels may present a challenge, but hope the experiment is do-able since data on m-Tyr incorporation into the proteome when cells are grown in m-Tyr are already provided.

Numerous attempts were made to address this question, but were unsuccessful; this is noted and explained in detail in the Results section.