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Myosin IIA regulates cell motility and actomyosin–microtubule crosstalk

Nature Cell Biology volume 9, pages 299–309 (2007)Cite this article

An Addendum to this article was published on 01 April 2007

Abstract

Non-muscle myosin II has diverse functions in cell contractility, cytokinesis and locomotion, but the specific contributions of its different isoforms have yet to be clarified. Here, we report that ablation of the myosin IIA isoform results in pronounced defects in cellular contractility, focal adhesions, actin stress fibre organization and tail retraction. Nevertheless, myosin IIA-deficient cells display substantially increased cell migration and exaggerated membrane ruffling, which was dependent on the small G-protein Rac1, its activator Tiam1 and the microtubule moter kinesin Eg5. Myosin IIA deficiency stabilized microtubules, shifting the balance between actomyosin and microtubules with increased microtubules in active membrane ruffles. When microtubule polymerization was suppressed, myosin IIB could partially compensate for the absence of the IIA isoform in cellular contractility, but not in cell migration. We conclude that myosin IIA negatively regulates cell migration and suggest that it maintains a balance between the actomyosin and microtubule systems by regulating microtubule dynamics.

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Figure 1: The steady-state balance between myosin IIA and microtubules is altered after in vitro wounding.
Figure 2: Genetic myosin IIA deficiency impairs formation of actin stress fibres and focal adhesions.
Figure 3: Myosin IIA depletion impairs contractility but increases migration rates.
Figure 4: Rac activation in myosin IIA-deficient cells.
Figure 5: Microtubule expansion in myosin IIA-deficient cells.
Figure 6: Disruption of microtubules restores contractility of myosin IIA-deficient cells.
Figure 7: Inhibition of Eg5 kinesin abolishes the increase in migration and membrane ruffling of myosin II-deficient cells.

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References

  1. Danowski, B. A. Fibroblast contractility and actin organization are stimulated by microtubule inhibitors. J. Cell Sci. 93, 255–266 (1989).

    CAS  PubMed  Google Scholar 

  2. Straight, A. F. et al. Dissecting temporal and spatial control of cytokinesis with a myosin II inhibitor. Science 299, 1743–1747 (2003).

    Article  CAS  Google Scholar 

  3. Gordon, S. R. & Staley, C. A. Role of the cytoskeleton during injury-induced cell migration in corneal endothelium. Cell Motil. Cytoskeleton 16, 47–57 (1990).

    Article  CAS  Google Scholar 

  4. Gupton, S. L. et al. Cell migration without a lamellipodium: translation of actin dynamics into cell movement mediated by tropomyosin. J. Cell Biol. 168, 619–631 (2005).

    Article  CAS  Google Scholar 

  5. Ponti, A., Machacek, M., Gupton, S. L., Waterman-Storer, C. M. & Danuser, G. Two distinct actin networks drive the protrusion of migrating cells. Science 305, 1782–1786 (2004).

    Article  CAS  Google Scholar 

  6. Abercrombie, M., Heaysman, J. E. & Pegrum, S. M. The locomotion of fibroblasts in culture. IV. Electron microscopy of the leading lamella. Exp. Cell Res. 67, 359–367 (1971).

    Article  CAS  Google Scholar 

  7. Svitkina, T. M., Verkhovsky, A. B., McQuade, K. M. & Borisy, G. G. Analysis of the actin-myosin II system in fish epidermal keratocytes: mechanism of cell body translocation. J. Cell Biol. 139, 397–415 (1997).

    Article  CAS  Google Scholar 

  8. Small, J. V. & Kaverina, I. Microtubules meet substrate adhesions to arrange cell polarity. Curr. Opin. Cell Biol. 15, 40–47 (2003).

    Article  CAS  Google Scholar 

  9. Gupton, S. L. & Waterman-Storer, C. M. Spatiotemporal feedback between actomyosin and focal-adhesion systems optimizes rapid cell migration. Cell 125, 1361–1374 (2006).

    Article  CAS  Google Scholar 

  10. Honer, B., Citi, S., Kendrick-Jones, J. & Jockusch, B. M. Modulation of cellular morphology and locomotory activity by antibodies against myosin. J. Cell Biol. 107, 2181–2189 (1988).

    Article  CAS  Google Scholar 

  11. Kovacs, M., Wang, F., Hu, A., Zhang, Y. & Sellers, J. R. Functional divergence of human cytoplasmic myosin II: kinetic characterization of the non-muscle IIA isoform. J. Biol. Chem. 278, 38132–38140 (2003).

    Article  CAS  Google Scholar 

  12. Conti, M. A., Even-Ram, S., Liu, C., Yamada, K. M. & Adelstein, R. S. Defects in cell adhesion and the visceral endoderm following ablation of nonmuscle myosin heavy chain II-A in mice. J. Biol. Chem. 279, 41263–41266 (2004).

    Article  CAS  Google Scholar 

  13. Tullio, A. N. et al. Nonmuscle myosin II-B is required for normal development of the mouse heart. Proc. Natl Acad. Sci. USA 94, 12407–12412 (1997).

    Article  CAS  Google Scholar 

  14. Thompson, R. F. & Langford, G. M. Myosin superfamily evolutionary history. Anat. Rec. 268, 276–289 (2002).

    Article  CAS  Google Scholar 

  15. Lo, C. M. et al. Nonmuscle myosin IIB is involved in the guidance of fibroblast migration. Mol. Biol. Cell 15, 982–989 (2004).

    Article  CAS  Google Scholar 

  16. Sandquist, J. C., Swenson, K. I., Demali, K. A., Burridge, K. & Means, A. R. Rho kinase differentially regulates phosphorylation of nonmuscle myosin II isoforms A and B during cell rounding and migration. J. Biol. Chem. 281, 35873–35883 (2006).

    Article  CAS  Google Scholar 

  17. Welch, M. P., Odland, G. F. & Clark, R. A. Temporal relationships of F-actin bundle formation, collagen and fibronectin matrix assembly, and fibronectin receptor expression to wound contraction. J. Cell Biol. 110, 133–145 (1990).

    Article  CAS  Google Scholar 

  18. Pankov, R. et al. A Rac switch regulates random versus directionally persistent cell migration. J. Cell Biol. 170, 793–802 (2005).

    Article  CAS  Google Scholar 

  19. Nishiya, N., Kiosses, W. B., Han, J. & Ginsberg, M. H. An α4 integrin–paxillin–Arf–GAP complex restricts Rac activation to the leading edge of migrating cells. Nature Cell Biol. 7, 343–352 (2005).

    Article  CAS  Google Scholar 

  20. Watanabe, T., Noritake, J. & Kaibuchi, K. Regulation of microtubules in cell migration. Trends Cell Biol. 15, 76–83 (2005).

    Article  CAS  Google Scholar 

  21. Helfman, D. M. et al. Caldesmon inhibits nonmuscle cell contractility and interferes with the formation of focal adhesions. Mol. Biol. Cell 10, 3097–3112 (1999).

    Article  CAS  Google Scholar 

  22. Gartner, M. et al. Development and biological evaluation of potent and specific inhibitors of mitotic kinesin Eg5. Chembiochem. 6, 1173–1177 (2005).

    Article  CAS  Google Scholar 

  23. Marcus, A. I. et al. Mitotic kinesin inhibitors induce mitotic arrest and cell death in Taxol-resistant and -sensitive cancer cells. J. Biol. Chem. 280, 11569–11577 (2005).

    Article  CAS  Google Scholar 

  24. Mertens, A. E., Roovers, R. C. & Collard, J. G. Regulation of Tiam1-Rac signalling. FEBS Lett. 546, 11–16 (2003).

    Article  CAS  Google Scholar 

  25. Katsumi, A. et al. Effects of cell tension on the small GTPase Rac. J. Cell Biol. 158,15 3–164 (2002).

    Article  Google Scholar 

  26. Meshel, A. S., Wei, Q., Adelstein, R. S. & Sheetz, M. P. Basic mechanism of three-dimensional collagen fibre transport by fibroblasts. Nature Cell Biol. 7, 157–164 (2005).

    Article  CAS  Google Scholar 

  27. Salmon, W. C., Adams, M. C. & Waterman-Storer, C. M. Dual-wavelength fluorescent speckle microscopy reveals coupling of microtubule and actin movements in migrating cells. J. Cell Biol. 158, 31–37 (2002).

    Article  CAS  Google Scholar 

  28. Medeiros, N. A., Burnette, D. T. & Forscher, P. Myosin II functions in actin-bundle turnover in neuronal growth cones. Nature Cell Biol. 8, 215–226 (2006).

    Article  CAS  Google Scholar 

  29. Waterman-Storer, C. M., Gregory, J., Parsons, S. F. & Salmon, E. D. Membrane/microtubule tip attachment complexes (TACs) allow the assembly dynamics of plus ends to push and pull membranes into tubulovesicular networks in interphase Xenopus egg extracts. J. Cell Biol. 130, 1161–1169 (1995).

    Article  CAS  Google Scholar 

  30. Kunda, P., Paglini, G., Quiroga, S., Kosik, K. & Caceres, A. Evidence for the involvement of Tiam1 in axon formation. J. Neurosci. 21, 2361–2372 (2001).

    Article  CAS  Google Scholar 

  31. Kaverina, I. et al. Tensile stress stimulates microtubule outgrowth in living cells. J. Cell Sci. 115, 2283–2291 (2002).

    CAS  PubMed  Google Scholar 

  32. Sawin, K. E., LeGuellec, K., Philippe, M. & Mitchison, T. J. Mitotic spindle organization by a plus-end-directed microtubule motor. Nature 359, 540–543 (1992).

    Article  CAS  Google Scholar 

  33. Yoon, S. Y. et al. Monastrol, a selective inhibitor of the mitotic kinesin Eg5, induces a distinctive growth profile of dendrites and axons in primary cortical neuron cultures. Cell Motil. Cytoskeleton 60, 181–190 (2005).

    Article  CAS  Google Scholar 

  34. Krylyshkina, O. et al. Modulation of substrate adhesion dynamics via microtubule targeting requires kinesin-1. J. Cell Biol. 156, 349–359 (2002).

    Article  CAS  Google Scholar 

  35. Conley, B. J. et al. Mouse embryonic stem cell derivation, and mouse and human embryonic stem cell culture and differentiation as embryoid bodies. Curr. Protocols Cell Biol. 23, 2.1–2.22 (2005).

    Google Scholar 

  36. Koivisto, L. et al. Glycogen synthase kinase-3 regulates cytoskeleton and translocation of Rac1 in long cellular extensions of human keratinocytes. Exp. Cell Res. 293, 68–80 (2004).

    Article  CAS  Google Scholar 

  37. Takeda, K., Yu, Z. X., Qian, S., Chin, T. K., Adelstein R. S. & Ferrans, V. J. Nonmuscle myosin II localizes to the Z-lines and intercalated discs of cardiac muscle and to the Z-lines of skeletal muscle. Cell Motil. Cytoskeleton 46, 59–68 (2000).

    Article  CAS  Google Scholar 

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Acknowledgements

This research was supported by the Intramural Research Program of the National Institutes of Health (NIH), National Institute of Dental and Craniofacial Research and National Heart, Lung, and Blood Institute.

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Authors and Affiliations

  1. Craniofacial Developmental Biology and Regeneration Branch, National Institute of Dental and Craniofacial Research (NIDCR), National Institutes of Health, Bethesda, 20892, MD, USA

    Sharona Even-Ram, Andrew D. Doyle, Kazue Matsumoto & Kenneth M. Yamada

  2. Laboratory of Molecular Cardiology, National Heart, Lung, and Blood Institute (NHLBI), National Institutes of Health, Bethesda, 20892, MD, USA

    Mary Anne Conti & Robert S. Adelstein

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  1. Sharona Even-Ram
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Correspondence to Kenneth M. Yamada.

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Even-Ram, S., Doyle, A., Conti, M. et al. Myosin IIA regulates cell motility and actomyosin–microtubule crosstalk. Nat Cell Biol 9, 299–309 (2007). https://doi.org/10.1038/ncb1540

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