Role of Rho, Rac, and Rho-kinase in phosphorylation of myosin light chain, development of polarity, and spontaneous migration of Walker 256 carcinosarcoma cells

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2005
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Gutjahr, Marc C.
Niggli, Verena
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Zusammenfassung

As previously shown, constitutive activation of the small GTPase Rho and its downstream target Rho-kinase is crucial for spontaneous migration of Walker carcinosarcoma cells.

We now show that after treatment of cells with either the Rho inhibitor C3 exoenzyme or the Rho-kinase inhibitor Y-27632, constitutive myosin light chain (MLC) phosphorylation is significantly decreased, correlating with inhibition of cell polarization and migration. Transfection with a dominant-negative Rho-kinase mutant similarly inhibits cell polarization and MLC phosphorylation. Transfection with a dominant-active Rho-kinase mutant leads to significantly increased MLC phosphorylation, membrane blebbing, and inhibition of cell polarization. This Rho-kinase-induced membrane blebbing can be inhibited by Y-27632, ML-7, and blebbistatin.

Unexpectedly, overactivation of RhoA has similar effects as its inhibition. Introduction of a bacterially expressed constitutively activated mutant protein (but not of wild-type RhoA) into the cells or transfection of cells with a constitutively active RhoA mutant both inhibit polarization and decrease MLC phosphorylation. Transfection of cells with constitutively active or dominant-negative Rac both abrogate polarity, and the latter inhibits MLC phosphorylation.

Our findings suggest an important role of Rac, Rho/Rho-kinase, and MLCK in controlling myosin activity in Walker carcinosarcoma cells and show that an appropriate level of RhoA, Rac, and Rho-kinase activity is required to regulate cell polarity and migration.

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570 Biowissenschaften, Biologie
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Walker carcinosarcoma cell, Rac, Rho, Rho-kinase, Myosin light chain, Myosin light chain kinase, Membrane blebbing, Cell migration
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ISO 690GUTJAHR, Marc C., Jérémie ROSSY, Verena NIGGLI, 2005. Role of Rho, Rac, and Rho-kinase in phosphorylation of myosin light chain, development of polarity, and spontaneous migration of Walker 256 carcinosarcoma cells. In: Experimental Cell Research. 2005, 308(2), pp. 422-438. ISSN 0014-4827. eISSN 1090-2422. Available under: doi: 10.1016/j.yexcr.2005.05.001
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@article{Gutjahr2005-08-15Rhoki-43215,
  year={2005},
  doi={10.1016/j.yexcr.2005.05.001},
  title={Role of Rho, Rac, and Rho-kinase in phosphorylation of myosin light chain, development of polarity, and spontaneous migration of Walker 256 carcinosarcoma cells},
  number={2},
  volume={308},
  issn={0014-4827},
  journal={Experimental Cell Research},
  pages={422--438},
  author={Gutjahr, Marc C. and Rossy, Jérémie and Niggli, Verena}
}
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