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Cdk1-dependent mitotic enrichment of cortical myosin II promotes cell rounding against confinement

Nature Cell Biology volume 17, pages 148159 (2015) | Download Citation

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Abstract

Actomyosin-dependent mitotic rounding occurs in both cell culture and tissue, where it is involved in cell positioning and epithelial organization. How actomyosin is regulated to mediate mitotic rounding is not well understood. Here we characterize the mechanics of single mitotic cells while imaging actomyosin recruitment to the cell cortex. At mitotic onset, the assembly of a uniform DIAPH1-dependent F-actin cortex coincides with initial rounding. Thereafter, cortical enrichment of F-actin remains stable while myosin II progressively accumulates at the cortex, and the amount of myosin at the cortex correlates with intracellular pressure. Whereas F-actin provides only short-term (<10 s) resistance to mechanical deformation, myosin sustains intracellular pressure for a longer duration (>60 s). Our data suggest that progressive accumulation of myosin II to the mitotic cell cortex probably requires the Cdk1 activation of both p21-activated kinases, which inhibit myosin recruitment, and of Rho kinase, which stimulates myosin recruitment to the cortex.

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Acknowledgements

We thank S. Weiser, Y. Toyoda, S. Raghuraman, R. Newton and B. Sorce for input, S. Hayashi, T. Kondo, M. Piel, C. Cabernard and O. Pertz for critical reading of the manuscript, the E. Paluch and F. Buchholz laboratories for contributing HeLa cell lines, J. Mercer for providing the mutant Rho GTPase plasmids as well as Cdc42 inhibitor, pirl1, and T. Horn and A. Ponti for help with the experiments on fluorescence recovery after photobleaching. This work was supported by the Eidgenössische Technische Hochschule Zürich (ETH research grant ETH-05 11-2), Swiss National Science Foundation (advanced mobility fellowship to M.P.S. and grant 31003A_138063 to D.J.M.) and National Center of Competence in Research Nano.

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Affiliations

  1. Eidgenössische Technische Hochschule (ETH) Zurich, Department of Biosystems Science and Engineering, 4058 Basel, Switzerland

    • Subramanian P. Ramanathan
    • , Jonne Helenius
    • , Martin P. Stewart
    • , Cedric J. Cattin
    •  & Daniel J. Muller
  2. Massachusetts Institute of Technology (MIT), Department of Chemical Engineering, Cambridge, Massachusetts 02139, USA

    • Martin P. Stewart
  3. The David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, Massachusetts 02139, USA

    • Martin P. Stewart
  4. Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany

    • Anthony A. Hyman

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Contributions

S.P.R., J.H., M.P.S., A.A.H. and D.J.M. planned the research and wrote the paper. S.P.R. designed, carried out and analysed most experiments. M.P.S. designed the STC-arrest methodology. M.P.S. and C.J.C. contributed to experiments in Supplementary Figs 7 and 8 respectively. J.H. wrote the scripts for image analysis.

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The authors declare no competing financial interests.

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Correspondence to Anthony A. Hyman or Daniel J. Muller.

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https://doi.org/10.1038/ncb3098

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