Peer review process
Revised: This Reviewed Preprint has been revised by the authors in response to the previous round of peer review; the eLife assessment and the public reviews have been updated where necessary by the editors and peer reviewers.
Read more about eLife’s peer review process.Editors
- Reviewing EditorAlan HinnebuschEunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, United States of America
- Senior EditorDetlef WeigelMax Planck Institute for Biology Tübingen, Tübingen, Germany
Reviewer #1 (Public Review):
The manuscript by Sejour et al. is testing "translational ramp" model described previously by Tuller et al. in S. cerevisiae. Authors are using bioinformatics and reporter based experimental approaches to test whether "rare codons" in the first 40 codons of the gene coding sequences increase translation efficiency and regulate abundance of translation products in yeast cells. Authors conclude that "translation ramp" model does not have support using a new set of reporters and bioinformatics analyses. The strength of bioinformatic evidence and experimental analyses (even very limited) of the rare codons insertion in the reporter make a compelling case for the authors claims. However the major weakness of the manuscript is that authors do not take into account other models that previously disputed "rare or slow codon" model of Tuller et al. and overstate their own results that are rather limited. This maintains to be the weak part of the manuscript even in the revised form.
The studies that authors do not mention argue with "translation ramp" model and show more thorough analyses of translation initiation to elongation transition as well as early elongation "slow down" in ribosome profiling data. Moreover several studies have used bioinformatical analyses to point out the evolution of N-terminal sequences in multiple model organisms including yeast, focusing on either upstream ORFs (uORFs) or already annotated ORFs. The authors did not mention multiple of these studies in their revised manuscript and did not comment on their own results in the context of these previous studies. As such the authors approach to data presentation, writing and data discussion makes the manuscript rather biased, focused on criticizing Tuller et al. study and short on discussing multiple other possible reasons for slow translation elongation at the beginning of the protein synthesis. This all together makes the manuscript at the end very limited.
Reviewer #2 (Public Review):
Tuller et al. first made the curious observation, that the first ∼30-50 codons in most organisms are encoded by scarce tRNAs and appear to be translated slower than the rest of the coding sequences (CDS). They speculated that this has evolved to pace ribosomes on CDS and prevent ribosome collisions during elongation - the "Ramp" hypothesis. Various aspects of this hypothesis, both factual and in terms of interpreting the results, have been challenged ever since. Sejour et al. present compelling results confirming the slower translation of the first ~40 codons in S. cerevisiae but providing an alternative explanation for this phenomenon. Specifically, they show that the higher amino acid sequence divergence of N-terminal ends of proteins and accompanying lower purifying selection (perhaps the result of de novo evolution) is sufficient to explain the prevalence of rare slow codons in these regions. These results are an important contribution in understanding how aspects of the evolution of protein coding regions can affect translation efficiency on these sequences and directly challenge the "Ramp" hypothesis proposed by Tuller et al.
I believe the data is presented clearly and the results generally justify the conclusions.