The crosstalk between microtubules, actin and membranes shapes cell division

Mitotic progression is orchestrated by morphological and mechanical changes promoted by the coordinated activities of the microtubule (MT) cytoskeleton, the actin cytoskeleton and the plasma membrane (PM). MTs assemble the mitotic spindle, which assists sister chromatid separation, and contact the rigid and tensile actomyosin cortex rounded-up underneath the PM. Here, we highlight the dynamic crosstalk between MTs, actin and cell membranes during mitosis, and discuss the molecular connections between them. We also summarize recent views on how MT traction forces, the actomyosin cortex and membrane trafficking contribute to spindle positioning in isolated cells in culture and in epithelial sheets. Finally, we describe the emerging role of membrane trafficking in synchronizing actomyosin tension and cell shape changes with cell–substrate adhesion, cell–cell contacts and extracellular signalling events regulating proliferation.

-Chaigne, A., Campillo, C., Voituriez, R., Gov, N 3/ I would cite this paper, an example of biochemical signaling between the cortex and the chromosomes, showing the coordination by Cdk1 of cortical tension maintenance and SAC inactivation at anaphase onset in mitotic cells.

Review form: Reviewer 2
Recommendation Accept with minor revision (please list in comments)

Do you have any ethical concerns with this paper? No
Comments to the Author The review paper by Rizzelli et al. entitled 'The crosstalk between microtubules, actin and membranes shape cell division' focuses on an interesting area to review. This paper presents an overview of current studies focusing on dynamics of the actin-and microtubule cytoskeleton and their link to the plasma membrane organization upon mitosis. Finally, the authors also highlight studies focusing on the relevance of these processes for cell-substrate and cell-cell adhesion as well as ECM signaling upon mitotic cell division and cytokinesis for the case of cells being embedded in epithelial sheets. Overall, the broader relevance of this nice review paper to the field of cell biology with a focus on mitotic cytoskeleton and adhesion is evident and my suggestions therefore only minor but may help to improve the reach of this paper, especially regarding figures and tables, as stated below. Abstract and Introduction The reader would benefit from getting a complete outline of the whole manuscript here including the three sections and ten subsections -if not provided by the journal anyway. All further comments are following this outline's structure. 1) Mitosis and cytoskeleton rearrangements 1.1 Actin and microtubule cytoskeleton in mitosis Citing thirteen paper in half a page seems a bit overwhelming, but with some more detail per paper -or less citations -it might be easier to follow the authors guide to the cited literature. For a general introduction, I would also recommend citing the paper by Dogterom and Koenderink on 'Actin-microtubule crosstalk in cell biology' for a non-mitotic view on the subject. 1.2 Adhesion in mitosis As the figures 1 A to C burst with information, I would recommend here to have one extra 'zoom in'/focus on the comparison of mitotic focal adhesion to the interphase adhesion and change the title also to adhesion in interphase and mitosis. Otherwise, it might be more instructive to cite figure 1 A and B already here, as the link from adhesion to spindle orientation becomes clear only in the latter figure but is already mentioned in the text. 1.3 Interplay between shape, the actomyosin cortex and spindle orientation The sentence containing first LGN mentioning is a bit misleading because of coma or missing parenthesis right before the word LGN. Also the general threefold subdivision in LGN and NuMA, actin clouds and ABP like myosin, dynein, ERM or Afdain might be prepared more by numbering or subsectioning components in figure 1 B. Again starting to mention the next topic from section 1.4, the MT motors, already in this subsection is a bit confusing (as with the spindle orientation in section 1.2 and 1.3). 1.4 Microtubule motors moving the mitotic spindle It might be enough to refer to Figure 1 C only in this subsection and the also show Gai only here and not already in figure 1B. On the other hand, the reader might benefit from a molecular explanation of the structure of NuMA or/and 4.1R proteins in advance, possibly by having another 'zoom in' in figure 1C on its interaction with Gai, dynactin and dynein. As the authors cite roughly 20 paper in sections 1.3. and 1.4, these resemble highlight parts and might therefore be reflected a bit more also by the figures.
2) Role of endocytosis in mitosis and cell division 2.1 Endocytic regulation of PM remodeling and mechanical forces In this chapter, there is no reference to a figure although I would suggest that a figure detailing the difference between Clathrin pits and plaques -especially in terms of integrins and mitosis relevance -would be helpful to understand the term 'frustrated endocytosis'. The same is valid for the distinction between CME and NCE including caveolae, thus resembling a figure 2A with a focus on the internalization routes at the top. The authors cite 35 paper is this subchapter, so an additional (sub)figure would be good to understand the context. 2.2 Role of endocytosis and trafficking in the regulation of PM remodeling during mitosis Besides internalization via CME, caveolae and NCE, recycling, (lysosome) exocytosis as well as membrane composition modulation are introduced here as cell surface are regulators. I would suggest to try to make a table for this very comprehensive subchapter (60 papers with 2.3) -the probably most central one in this review -summarizing some of the central results found for the actors like the different Rab proteins, but possibly also some of the actors mentioned in section 2.3. 2.3 Endocytic proteins with functions in mitosis and cytokinesis See above, point 2.2 3) Cell division and epithelial dynamics: the role of AJs and their regulation by endocytosis 3.1 AJs are critical sensors of forces in polarized epithelia A more detailed exemplary scheme of the e-cadherine might be show in figure 3 A but is not absolutely relevant for the reader to understand the dynamics summarized in section 3. However, I would include the part on EMT as one example here and only start the next subsection with the sentence including literature citations 168 and 171, as the mitotic focus becomes more obvious here. 3.2 The role of endocytosis in AJ remodeling during epithelial cell division See above. There are some formatting issues before citation 218. EE and LE is not defined in figure 3 A nor in the caption. 3.3 Interaction of mitotic cells with the extracellular matrix: unexpected link between mitotic focal adhesion and endocytic plaques I suggest to label the reticular adhesion in figure 3 B with the abbreviation RA introduced in the text.

Dear Dr Mapelli
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Data accessibility section To ensure archived data are available to readers, authors should include a 'data accessibility' section immediately after the acknowledgements section. This should list the database and accession number for all data from the article that has been made publicly available, for instance: Comments to the Author(s) This review discusses the current knowledge about the molecular mechanisms at the interplay between the actomyosin cortex, the microtubule spindle and membrane trafficking in the coordination of mitotic progression. This review is well written and well documented, spanning old and new bibliography, a rarity nowadays where reviews usually cover only the last 5 to 10 years. It goes deep in the understanding of the processes and is not just a list of papers.
I have only very few comments to improve it: 1/ I would reinforce the part on the interplay between actin and microtubules (end of page 6). The authors cite the paper describing the ability of centrosomes to nucleate actin (Farina EMBO J 2019), but they should also cite these very recent other papers highlighting the fact F-actin is known to impact microtubule architecture and dynamics, potentially acting on chromosome behavior ( Comments to the Author(s) The review paper by Rizzelli et al. entitled 'The crosstalk between microtubules, actin and membranes shape cell division' focuses on an interesting area to review. This paper presents an overview of current studies focusing on dynamics of the actin-and microtubule cytoskeleton and their link to the plasma membrane organization upon mitosis. Finally, the authors also highlight studies focusing on the relevance of these processes for cell-substrate and cell-cell adhesion as well as ECM signaling upon mitotic cell division and cytokinesis for the case of cells being embedded in epithelial sheets. Overall, the broader relevance of this nice review paper to the field of cell biology with a focus on mitotic cytoskeleton and adhesion is evident and my suggestions therefore only minor but may help to improve the reach of this paper, especially regarding figures and tables, as stated below.

Abstract and Introduction
The reader would benefit from getting a complete outline of the whole manuscript here including the three sections and ten subsections -if not provided by the journal anyway. All further comments are following this outline's structure. 1) Mitosis and cytoskeleton rearrangements 1.1 Actin and microtubule cytoskeleton in mitosis Citing thirteen paper in half a page seems a bit overwhelming, but with some more detail per paper -or less citations -it might be easier to follow the authors guide to the cited literature. For a general introduction, I would also recommend citing the paper by Dogterom and Koenderink on 'Actin-microtubule crosstalk in cell biology' for a non-mitotic view on the subject. 1.2 Adhesion in mitosis As the figures 1 A to C burst with information, I would recommend here to have one extra 'zoom in'/focus on the comparison of mitotic focal adhesion to the interphase adhesion and change the title also to adhesion in interphase and mitosis. Otherwise, it might be more instructive to cite figure 1 A and B already here, as the link from adhesion to spindle orientation becomes clear only in the latter figure but is already mentioned in the text. 1.3 Interplay between shape, the actomyosin cortex and spindle orientation The sentence containing first LGN mentioning is a bit misleading because of coma or missing parenthesis right before the word LGN. Also the general threefold subdivision in LGN and NuMA, actin clouds and ABP like myosin, dynein, ERM or Afdain might be prepared more by numbering or subsectioning components in figure 1 B. Again starting to mention the next topic from section 1.4, the MT motors, already in this subsection is a bit confusing (as with the spindle orientation in section 1.2 and 1.3). 1.4 Microtubule motors moving the mitotic spindle It might be enough to refer to Figure 1 C only in this subsection and the also show Gai only here and not already in figure 1B. On the other hand, the reader might benefit from a molecular explanation of the structure of NuMA or/and 4.1R proteins in advance, possibly by having another 'zoom in' in figure 1C on its interaction with Gai, dynactin and dynein. As the authors cite roughly 20 paper in sections 1.3. and 1.4, these resemble highlight parts and might therefore be reflected a bit more also by the figures.
2) Role of endocytosis in mitosis and cell division 2.1 Endocytic regulation of PM remodeling and mechanical forces In this chapter, there is no reference to a figure although I would suggest that a figure detailing the difference between Clathrin pits and plaques -especially in terms of integrins and mitosis relevance -would be helpful to understand the term 'frustrated endocytosis'. The same is valid for the distinction between CME and NCE including caveolae, thus resembling a figure 2A with a focus on the internalization routes at the top. The authors cite 35 paper is this subchapter, so an additional (sub)figure would be good to understand the context.

Role of endocytosis and trafficking in the regulation of PM remodeling during mitosis
Besides internalization via CME, caveolae and NCE, recycling, (lysosome) exocytosis as well as membrane composition modulation are introduced here as cell surface are regulators. I would suggest to try to make a table for this very comprehensive subchapter (60 papers with 2.3) -the probably most central one in this review -summarizing some of the central results found for the actors like the different Rab proteins, but possibly also some of the actors mentioned in section 2.3. 2.3 Endocytic proteins with functions in mitosis and cytokinesis See above, point 2.2 3) Cell division and epithelial dynamics: the role of AJs and their regulation by endocytosis 3.1 AJs are critical sensors of forces in polarized epithelia A more detailed exemplary scheme of the e-cadherine might be show in figure 3 A but is not absolutely relevant for the reader to understand the dynamics summarized in section 3. However, I would include the part on EMT as one example here and only start the next subsection with the sentence including literature citations 168 and 171, as the mitotic focus becomes more obvious here. 3.2 The role of endocytosis in AJ remodeling during epithelial cell division See above. There are some formatting issues before citation 218. EE and LE is not defined in figure 3 A nor in the caption.

Interaction of mitotic cells with the extracellular matrix:
unexpected link between mitotic focal adhesion and endocytic plaques I suggest to label the reticular adhesion in figure 3 B with the abbreviation RA introduced in the text.

18-Feb-2020 Dear Dr Mapelli
We are pleased to inform you that your manuscript entitled "The crosstalk between microtubules, actin and membranes shapes cell division" has been accepted by the Editor for publication in Open Biology.
You can expect to receive a proof of your article from our Production office in due course, please check your spam filter if you do not receive it within the next 10 working days. Please let us know if you are likely to be away from e-mail contact during this time.
Thank you for your fine contribution. On behalf of the Editors of Open Biology, we look forward to your continued contributions to the journal. Dear Dr. Glover, please find enclosed the response to the Reviewers' comments to the manuscript 'The crosstalk between microtubules, actin and membranes shape cell division', which was assessed by Open Biology Referees (RSOB-19-0314).
We have revised the text and the figures of the manuscript according to the Referees' suggestions, as detailed below in the point-by-point answers to the Referees' comments. I thank you in advance for the editorial help and the patience, and I look forward to hearing from you.

With my kindest regards, Marina Mapelli
Reviewer(s)' Comments to Author:

Referee:
1 Comments to the Author(s) This review discusses the current knowledge about the molecular mechanisms at the interplay between the actomyosin cortex, the microtubule spindle and membrane trafficking in the coordination of mitotic progression. This review is well written and well documented, spanning old and new bibliography, a rarity nowadays where reviews usually cover only the last 5 to 10 years. It goes deep in the understanding of the processes and is not just a list of papers.
I have only very few comments to improve it: 1/ I would reinforce the part on the interplay between actin and microtubules (end of page 6). The authors cite the paper describing the ability of centrosomes to nucleate actin (Farina EMBO J 2019), but they should also cite these very recent other papers highlighting the fact F-actin is known to impact microtubule architecture and dynamics, potentially acting on chromosome behavior (Chaigne et al., 2016;Colin et al., 2018;Inoue et al., 2019;Farina et al., 2019;Kita et al., 2019;Plessner et al., 2019). We thank the Reviewer for the positive comments and the suggestions. We have added the mentioned references and strengthened the description of the mitotic interplay between actin and microtubules in the first section.
We have added a sentence describing the role of myosin II in modulating MCAK-dependent microtubule growth in mitosis, and cited the corresponding reference (see page 6).
3/ I would cite this paper, an example of biochemical signaling between the cortex and the chromosomes, showing the coordination by Cdk1 of cortical tension maintenance and SAC inactivation at anaphase onset in mitotic cells.
We thank the Referee for the suggestion. We have added a sentence describing the role of the Cdk1-dependent phosphorylation of DIAPH1 in ensuing and maintaining cortical tension during mitotic round up and cited the associated reference (page 4). 4/ At last, please replace Bornes and Thiery end of page 5 by Bornens and Thery.
We have corrected the typo.

Referee-2:
Comments to the Author(s) The review paper by Rizzelli et al. entitled 'The crosstalk between microtubules, actin and membranes shape cell division' focuses on an interesting area to review. This paper presents an overview of current studies focusing on dynamics of the actin-and microtubule cytoskeleton and their link to the plasma membrane organization upon mitosis. Finally, the authors also highlight studies focusing on the relevance of these processes for cell-substrate and cell-cell adhesion as well as ECM signaling upon mitotic cell division and cytokinesis for the case of cells being embedded in epithelial sheets. Overall, the broader relevance of this nice review paper to the field of cell biology with a focus on mitotic cytoskeleton and adhesion is evident and my suggestions therefore only minor but may help to improve the reach of this paper, especially regarding figures and tables, as stated below.

Abstract and Introduction
The reader would benefit from getting a complete outline of the whole manuscript here including the three sections and ten subsections -if not provided by the journal anyway. All further comments are following this outline's structure.
We thank the Reviewer for the suggestion. We have added a paragraph at the end of the Introduction to outline the organization of the Review in three parts, indicating the focus of the each of them. However, we have preferred to omit the list of sub-section to avoid being too detailed.

1)
Mitosis and cytoskeleton rearrangements 1.1 Actin and microtubule cytoskeleton in mitosis Citing thirteen paper in half a page seems a bit overwhelming, but with some more detail per paper -or less citations -it might be easier to follow the authors guide to the cited literature. For a general introduction, I would also recommend citing the paper by Dogterom and Koenderink on 'Actin-microtubule crosstalk in cell biology' for a non-mitotic view on the subject.
We agree with the Referee that the paragraph describing the actin-microtubule crosstalk is rather dense, and contains a substantial number of citations. We have better described the actin-dependent rounding mechanisms (including the role of DIAPH1), and cited the review by Dogterom and Koenderink for a comprehensive description of the current knowledge on the actin-microtubule cross-talk.

1.2
Adhesion in mitosis As the figures 1 A to C burst with information, I would recommend here to have one extra 'zoom in'/focus on the comparison of mitotic focal adhesion to the interphase adhesion and change the title also to adhesion in interphase and mitosis. Otherwise, it might be more instructive to cite figure 1 A and B already here, as the link from adhesion to spindle orientation becomes clear only in the latter figure but is already mentioned in the text.
We take the Referee's point that in the paragraph "Adhesion in mitosis" we provide a snapshot of adhesion mechanisms, including interphase and mitotic adhesion complexes, that is very concise. Our decision to adopt this perspective stems from the idea that the focus of the Review is to describe changes between interphase and mitosis, rather than to provide an exhaustive description of the two cell cycle phases. Also, we are mentioning adhesion complexes mainly to connect them to the spindle positioning mechanisms described in Figure  1B. Following the Referee suggestion, to make the text clearer to the readers, we have cited Figure 1A and 1B directly in this "Adhesion in mitosis" paragraph.

1.3
Interplay between shape, the actomyosin cortex and spindle orientation The sentence containing first LGN mentioning is a bit misleading because of coma or missing parenthesis right before the word LGN.
We have corrected the typo and removed the extra comma.
Also the general threefold subdivision in LGN and NuMA, actin clouds and ABP like myosin, dynein, ERM or Afadin might be prepared more by numbering or subsectioning components in figure 1 B.
We agree with the Reviewer. We have added dashed boxes to the Figure 1B, highlighting the modules cited by the Referee, i.e. ERM-containing mitotic focal adhesion complexes, actin clouds, and actin binding proteins, and cited them appropriately in the main text. In addition, we have added a separate close-up view of the LGN/NuMA module in an additional BOX-1.
Again starting to mention the next topic from section 1.4, the MT motors, already in this subsection is a bit confusing (as with the spindle orientation in section 1.2 and 1.3).
We thank the Referee for the comment. Our decision to start mentioning MT-motors in this paragraph reflects the notion that actin-binding proteins are implicated in mitotic cortical organization as well as in spindle placement. Thus, we preferred to introduce them already in this paragraph, and describe in detail their functions in the dedicated paragraph "Microtubule motors moving the mitotic spindle".

1.4
Microtubule motors moving the mitotic spindle It might be enough to refer to Figure 1 C only in this subsection and the also show Gai only here and not already in figure 1B. On the other hand, the reader might benefit from a molecular explanation of the structure of NuMA or/and 4.1R proteins in advance, possibly by having another 'zoom in' in figure 1C on its interaction with Gai, dynactin and dynein. As the authors cite roughly 20 paper in sections 1.3. and 1.4, these resemble highlight parts and might therefore be reflected a bit more also by the figures.
We thank the Referee for the suggestions. Gai molecules are implicated in recruiting LGN and NuMA at the cortex from metaphase on, therefore we prefer to leave them also in Figure  1B rather than introducing them only in Figure 1C. We agree that the description of the properties of NuMA and LGN only in the text and references may be difficult to visualize in the context of the cartoons shown in Figure 1B-C. To facilitate the readers, we have added a BOX-1 with a close-up view of Gai/LGN/NuMA interactions, and a schematic representation of the domain structure of NuMA and LGN annotated with all the interactions mentioned in the text and the relative reference.

2)
Role of endocytosis in mitosis and cell division 2.1 Endocytic regulation of PM remodeling and mechanic forces.
In this chapter, there is no reference to a figure although I would suggest that a figure detailing the difference between Clathrin pits and plaques -especially in terms of integrins and mitosis relevance -would be helpful to understand the term 'frustrated endocytosis'. The same is valid for the distinction between CME and NCE including caveolae, thus resembling a figure 2A with a focus on the internalization routes at the top. The authors cite 35 paper is this subchapter, so an additional (sub)figure would be good to understand the context.
We agree with the Reviewer. We have added a new Figure, Figure 2, describing the different entry routes discussed in the text, with a focus to the distinction between clathrin-coated pits and plaques.

Role of endocytosis and trafficking in the regulation of PM remodeling during mitosis
Besides internalization via CME, caveolae and NCE, recycling, (lysosome) exocytosis as well as membrane composition modulation are introduced here as cell surface are regulators. I would suggest to try to make a table for this very comprehensive subchapter (60 papers with 2.3) -the probably most central one in this review -summarizing some of the central results found for the actors like the different Rab proteins, but possibly also some of the actors mentioned in section 2.3.
We thank the Reviewer for the suggestion. We have added a new Table I, summarizing the different endocytic proteins that have been discussed along the text.

Endocytic proteins with functions in mitosis and cytokinesis
See above, point 2.2 3) Cell division and epithelial dynamics: the role of AJs and their regulation by endocytosis 3.1 AJs are critical sensors of forces in polarized epithelia A more detailed exemplary scheme of the e-cadherine might be show in figure 3 A but is not absolutely relevant for the reader to understand the dynamics summarized in section 3. However, I would include the part on EMT as one example here and only start the next subsection with the sentence including literature citations 168 and 171, as the mitotic focus becomes more obvious here.
We thank the Reviewer for the suggestion. We have rearranged the text by adding a new subsection, which starts with the sentence including literature citation 168 and 171 (now 178-181), to better highlight the mitotic focus of this paragraph.
3.2 The role of endocytosis in AJ remodeling during epithelial cell division See above. There are some formatting issues before citation 218.
We have solved the formatting issue.
EE and LE is not defined in figure 3 A nor in the caption.
We apologize for not having clarified this in the previous version of the manuscript. We have now explained the acronyms in the figure legend (now Figure 4A).
3.3 Interaction of mitotic cells with the extracellular matrix: unexpected link between mitotic focal adhesion and endocytic plaques. I suggest to label the reticular adhesion in figure 3 B with the abbreviation RA introduced in the text.
As it is not yet completely established whether all these structures (e.g. the mitotic-resistant adhesion complexes, the reticular adhesions, the coated plaques) are indeed the same type of structures, or specialized subtypes enriched in specific regions of the cell and/or in specific cellular contexts, we have preferred to refer to these structures more generically as 'mitotic focal adhesion complexes'. We have however added a sentence to explain better their relationship in the legend of Figure 4.