Dear Dr Tewari,

Thank you for submitting your manuscript entitled "Key roles for kinesin-13 and kinesin-20 in malaria parasite proliferation, polarity, and transmission revealed by genome-wide functional analysis" for consideration as a Research Article by PLOS Biology.

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Kind regards,

Dario

Dario Ummarino, PhD

Senior Editor

PLOS Biology

dummarino@plos.org

Dear Dr Tewari,

Thank you for your patience while we considered your revised manuscript "Key roles for kinesin-13 and kinesin-20 in malaria parasite proliferation, polarity, and transmission revealed by genome-wide functional analysis" for publication as a Research Article at PLOS Biology. This revised version of your manuscript has been evaluated by the PLOS Biology editors, the Academic Editor and one of the original reviewers. One of the reviewers who originally reviewed your manuscript at Review Commons (reviewer 3) was not available for re-review, therefore the Academic Editor assessed directly your responses to their original concerns.

Based on the reviews and the Academic Editor's assessment of your revision, we are likely to accept this manuscript for publication, provided you satisfactorily address the remaining points raised by the reviewers and Academic Editors, which are pasted at the end of this email. Please also make sure to address the following data and other policy-related requests:

1) Title: We would like to suggest a modification for clarity and appeal: "Genome-wide functional analysis reveals key roles for kinesins in the mammalian and mosquito stages of the malaria parasite life cycle".

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Sincerely,

Dario

Dario Ummarino, PhD

Senior Editor

PLOS Biology

dummarino@plos.org

------------------------------------------------------------------------

Reviewer's report:

Reviewer #1: The authors addressed all comments from the Review Commons published in December 2021.

I have one last request upon endorsement for publication is to remove the term "endomitotic" in line 87.

Endomitosis means that the number of chromosomes increases without nuclear division, leading to the polyploid stage. In contrast, the Plasmodium parasite replicates by schizogony, where DNA replication is followed by nuclear division (PMID: 35353560). For instance, Sarcocystis replicates by endomitosis, not Plasmodium (suggested references are PMID: 17604449 and 33912479).

Understanding the function of microtubule motors in Plasmodium spp will advance our knowledge of the atypical cell division mode of the malaria parasite and unveil innovative ways to block the parasite replication.

Comments from the Academic Editor:

Point 1. Details are missing in the methods for how the authors performed the immunoprecipitation with the GFP-TRAP beads and there is also no mention of what served as the negative control. Moreover, the peptide counts in the negative controls should also be included in S2 Fig so we can see what the background counts are like.

Also regarding GFP expression, Fig 1B and 1C don’t completely talk to one another. Because of the large number of proteins they are looking at, they show 1B for all the various localisations and then then 1C as a summative of what they saw (for which we don't see the raw data). In Fig 1B, The GFP signal either localises to the Spindle Pole or Spindle in male gametocytes and it would be good to know which kinesins these were. There are also no pictures of female gametocytes, male gametes or zygotes in Fig 1 B. Whilst it has been previously published that Kinesin 5 and 8X co-localise with anti-tubulin antibodies (to detect spindle), I think it is important to show a couple of co-localisation figures in this manuscript since the focus is on spindles and the reader should not have to go to another publication to see what that labelling looks like.

Point 2. Chip-Seq data is presented in S7Fig and includes positive and negative controls as requested. However, information regarding the controls should also be in the methodology section.

Point 3. Whilst the authors have now made a change in the text regarding why they undertook RNAseq analysis on the gam stage, they didn't actually point out that their transcriptomic data showed no role for kinesin-20 in translational repression. This should be specified as it is simply inferred but not overly clear. Also line349: comma should be replaced with full stop. This suggests a link…..

Dear Dr Tewari,

Thank you for the submission of your revised Research Article "Genome-wide functional analysis reveals key roles for kinesins in the mammalian and mosquito stages of the malaria parasite life cycle" for publication in PLOS Biology. On behalf of my colleagues and the Academic Editor, Tania de Koning-Ward, I am pleased to say that we can in principle accept your manuscript for publication, provided you address any remaining formatting and reporting issues. These will be detailed in an email you should receive within 2-3 business days from our colleagues in the journal operations team; no action is required from you until then. Please note that we will not be able to formally accept your manuscript and schedule it for publication until you have completed any requested changes.

Please take a minute to log into Editorial Manager at http://www.editorialmanager.com/pbiology/, click the "Update My Information" link at the top of the page, and update your user information to ensure an efficient production process.

PRESS

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We also ask that you take this opportunity to read our Embargo Policy regarding the discussion, promotion and media coverage of work that is yet to be published by PLOS. As your manuscript is not yet published, it is bound by the conditions of our Embargo Policy. Please be aware that this policy is in place both to ensure that any press coverage of your article is fully substantiated and to provide a direct link between such coverage and the published work. For full details of our Embargo Policy, please visit http://www.plos.org/about/media-inquiries/embargo-policy/.

Thank you again for choosing PLOS Biology for publication and supporting Open Access publishing. We look forward to publishing your study. 

Sincerely, 

Dario

Dario Ummarino, PhD

Senior Editor

PLOS Biology