Peer review in Bub3 reads phosphorylated MELT repeats to promote spindle assembly checkpoint signaling

Decision letter
Author response

Decision letter

Jon Pines

Reviewing Editor; The Gurdon Institute, United Kingdom

eLife posts the editorial decision letter and author response on a selection of the published articles (subject to the approval of the authors). An edited version of the letter sent to the authors after peer review is shown, indicating the substantive concerns or comments; minor concerns are not usually shown. Reviewers have the opportunity to discuss the decision before the letter is sent (see review process). Similarly, the author response typically shows only responses to the major concerns raised by the reviewers.

Thank you for submitting your work entitled “Bub3 reads phosphorylated MELT repeats to promote spindle assembly checkpoint signaling” for consideration at eLife. Your article has been favorably evaluated by a Senior editor and three reviewers, one of whom is a member of the Board of Reviewing Editors.

One referee would like you to perform the following experiments to support your conclusions:

1) Arg239 that contacts pThr is a Lys in most Bub1 orthologs. Did the authors test whether a Lys substitution can still bind MELT peptides?

2) The authors suggest that Arg314 mediates the Bub1-dependent increased affinity of Bub3 for MELT peptides. What happens to the Bub1-mediated increase in binding when Arg314 is mutated?

https://doi.org/10.7554/eLife.01030.023

Author response

1) Arg239 that contacts pThr is a Lys in most Bub1 orthologs. Did the authors test whether a Lys substitution can still bind MELT peptides?

[Editors' note: at the authors' request, the answer to this question has not been published to avoid disclosing confidential information.]

2) The authors suggest that Arg314 mediates the Bub1-dependent increased affinity of Bub3 for MELT peptides. What happens to the Bub1-mediated increase in binding when Arg314 is mutated?

This was an interesting suggestion. We have carried out an additional calorimetry experiment with the Bub1^R314A mutant. In agreement with the hypothesis that this residue contributes to recognition of the phosphorylated MELT peptide, we find that the R314A mutant binds the MELT2^P peptide with ∼3- to 4-fold reduced affinity. We have included the new ITC data as Figure 4F.

https://doi.org/10.7554/eLife.01030.024