https://orcid.org/0000-0001-5094-8105
Peer Review History
| Original SubmissionSeptember 13, 2020 |
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Transfer Alert
This paper was transferred from another journal. As a result, its full editorial history (including decision letters, peer reviews and author responses) may not be present.
Dear Dr. Skaar, Thank you very much for submitting your manuscript "Directed evolution reveals the mechanism of HitRS signaling transduction in Bacillus anthracis" for consideration at PLOS Pathogens. As with all papers reviewed by the journal, your manuscript was reviewed by members of the editorial board and by several independent reviewers. The reviewers appreciated the attention to an important topic. Based on the reviews, we are likely to accept this manuscript for publication, providing that you modify the manuscript according to the review recommendations. Please prepare and submit your revised manuscript within 30 days. If you anticipate any delay, please let us know the expected resubmission date by replying to this email. When you are ready to resubmit, please upload the following: [1] A letter containing a detailed list of your responses to all review comments, and a description of the changes you have made in the manuscript. Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out [2] Two versions of the revised manuscript: one with either highlights or tracked changes denoting where the text has been changed; the other a clean version (uploaded as the manuscript file). Important additional instructions are given below your reviewer comments. Thank you again for your submission to our journal. We hope that our editorial process has been constructive so far, and we welcome your feedback at any time. Please don't hesitate to contact us if you have any questions or comments. Sincerely, Theresa M. Koehler Associate Editor PLOS Pathogens Michael Wessels Section Editor PLOS Pathogens Kasturi Haldar Editor-in-Chief PLOS Pathogens orcid.org/0000-0001-5065-158X Michael Malim Editor-in-Chief PLOS Pathogens *********************** Reviewer Comments (if any, and for reference): Reviewer's Responses to Questions Part I - Summary Please use this section to discuss strengths/weaknesses of study, novelty/significance, general execution and scholarship. Reviewer #1: In this study, Pi et al use a clever genetic design to perform a functional analysis of HitSR, a two-component system (TCS) of Bacillus anthracis (Ba). They isolate constitutively active or inactive variants. They generate and purify representative recombinant variants using E. coli for biochemical characterization of autokinase, phosphatase, phosphotransfer and DNA binding activities. The authors identify residues essential for each activity and for interaction between subdomains as well as between HitS and HitR. Some of the results are “as expected”. The design for selection of mutants can be generalized to the study of other TCS. Data and results are extensively explained, well-presented and conclusive. While this particular TCS belongs to a pathogen, the study is strictly limited to the structure-function analysis of the HitRS and does not investigate an impact on the pathogenesis of B. anthracis nor an impact on antibiotic resistance. Reviewer #2: The manuscript by Hualiang et al. characterizes the HitRS two component system, which has been implicated in detecting cell envelope stress, in Bacillus anthracis. The authors devised two genetic selections performed in B. anthracis to identify either critical residues in HitRS or constitutively active forms of HitRS. This strategy revealed gain-of-function mutations in the sensor domain and TM region of HitS and critical residues in the HAMP domain of HitS. The authors also report the in vivo consequence for B. anthracis growth in cells harboring various isolated HitRS alleles, examine protein stability and dimerization potential of the various mutants, and identify residues that are critical for HitR-HitS interaction. ********** Part II – Major Issues: Key Experiments Required for Acceptance Please use this section to detail the key new experiments or modifications of existing experiments that should be absolutely required to validate study conclusions. Generally, there should be no more than 3 such required experiments or major modifications for a "Major Revision" recommendation. If more than 3 experiments are necessary to validate the study conclusions, then you are encouraged to recommend "Reject". Reviewer #1: l.87-88: the authors cite compound VU0120205 ‘205 as a cell-envelope acting compound but the associated reference #23 describes compound VU0038882 (‘882) that activates HssRS by inducing endogenous heme biosynthesis in S. aureus. I wonder if the authors could clarify the relationship between HssRS and HitRS. I am not sure how responding to heme availability (HssRS) fits with sensing cell envelope perturbations (HitRS). Also, the authors should explain why they picked HitRS over HssRS. Reviewer #2: (No Response) ********** Part III – Minor Issues: Editorial and Data Presentation Modifications Please use this section for editorial suggestions as well as relatively minor modifications of existing data that would enhance clarity. Reviewer #1: l. 28: why is Ba referred as an intracellular pathogen? l. 36: this is a very strong statement. I don’t think that the study comes close to laying such a foundation. The “Author Summary” section uses slightly different words but essentially repeats the abstract. l.49: it is odd to start the paper with a statement about the rising incidence of antibiotic resistance. That is not a trait associated with Ba. Word on the street is that this pathogen is dormant in soils; mostly! Certainly, Ba isn’t roaming around hospitals with the Enterococci and the likes of them. l.58: during infection Ba is only transiently intracellular. Fig. 1 and corresponding Result section present clever approaches to isolate hit alleles with constitutive activity and mutations that inactivate signaling or signal transduction. However, this does not bring the authors closer to identifying the signal that activates HitRS. As expected mutations arising from these selections bypass the requirement for the true ligand of HitRS. Reviewer #2: In general, the experiments were carefully controlled, and the manuscript was well-written so that it should be accessible to non-experts. This was a straightforward but exhaustive characterization of a two-component system in an important pathogen. As such, it should be of interest to the broad signal transduction community and to those who study bacterial pathogenesis. I have no major scientific comments and only one minor cosmetic comment that the authors may or may not wish to incorporate. Minor comment: 1. Fig. S3. This schematic represents a major set of results of this study. Consider moving it to the main text (perhaps in Fig. 1?). ********** PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No Reviewer #2: No Figure Files: While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email us at figures@plos.org. Data Requirements: Please note that, as a condition of publication, PLOS' data policy requires that you make available all data used to draw the conclusions outlined in your manuscript. Data must be deposited in an appropriate repository, included within the body of the manuscript, or uploaded as supporting information. This includes all numerical values that were used to generate graphs, histograms etc.. For an example see here: http://www.plosbiology.org/article/info%3Adoi%2F10.1371%2Fjournal.pbio.1001908#s5. Reproducibility: To enhance the reproducibility of your results, PLOS recommends that you deposit laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. For instructions see http://journals.plos.org/plospathogens/s/submission-guidelines#loc-materials-and-methods |
| Revision 1 |
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Dear Dr. Skaar, We are pleased to inform you that your manuscript 'Directed evolution reveals the mechanism of HitRS signaling transduction in Bacillus anthracis' has been provisionally accepted for publication in PLOS Pathogens. Before your manuscript can be formally accepted you will need to complete some formatting changes, which you will receive in a follow up email. A member of our team will be in touch with a set of requests. Please note that your manuscript will not be scheduled for publication until you have made the required changes, so a swift response is appreciated. IMPORTANT: The editorial review process is now complete. PLOS will only permit corrections to spelling, formatting or significant scientific errors from this point onwards. Requests for major changes, or any which affect the scientific understanding of your work, will cause delays to the publication date of your manuscript. Should you, your institution's press office or the journal office choose to press release your paper, you will automatically be opted out of early publication. We ask that you notify us now if you or your institution is planning to press release the article. All press must be co-ordinated with PLOS. Thank you again for supporting Open Access publishing; we are looking forward to publishing your work in PLOS Pathogens. Best regards, Theresa M. Koehler Associate Editor PLOS Pathogens Michael Wessels Section Editor PLOS Pathogens Kasturi Haldar Editor-in-Chief PLOS Pathogens orcid.org/0000-0001-5065-158X Michael Malim Editor-in-Chief PLOS Pathogens *********************************************************** Reviewer Comments (if any, and for reference): |
| Formally Accepted |
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Dear Dr. Skaar, We are delighted to inform you that your manuscript, "Directed evolution reveals the mechanism of HitRS signaling transduction in Bacillus anthracis," has been formally accepted for publication in PLOS Pathogens. We have now passed your article onto the PLOS Production Department who will complete the rest of the pre-publication process. All authors will receive a confirmation email upon publication. The corresponding author will soon be receiving a typeset proof for review, to ensure errors have not been introduced during production. Please review the PDF proof of your manuscript carefully, as this is the last chance to correct any scientific or type-setting errors. Please note that major changes, or those which affect the scientific understanding of the work, will likely cause delays to the publication date of your manuscript. Note: Proofs for Front Matter articles (Pearls, Reviews, Opinions, etc...) are generated on a different schedule and may not be made available as quickly. Soon after your final files are uploaded, the early version of your manuscript, if you opted to have an early version of your article, will be published online. The date of the early version will be your article's publication date. The final article will be published to the same URL, and all versions of the paper will be accessible to readers. Thank you again for supporting open-access publishing; we are looking forward to publishing your work in PLOS Pathogens. Best regards, Kasturi Haldar Editor-in-Chief PLOS Pathogens orcid.org/0000-0001-5065-158X Michael Malim Editor-in-Chief PLOS Pathogens |
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