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Rac regulates phosphorylation of the myosin-II heavy chain, actinomyosin disassembly and cell spreading

Nature Cell Biology volume 1, pages 242–248 (1999)Cite this article

Abstract

GTPases of the Rho family regulate actinomyosin-based contraction in non-muscle cells. Activation of Rho increases contractility, leading to cell rounding and neurite retraction in neuronal cell lines. Activation of Rac promotes cell spreading and interferes with Rho-mediated cell rounding. Here we show that activation of Rac may antagonize Rho by regulating phosphorylation of the myosin-II heavy chain. Stimulation of PC12 cells or N1E-115 neuroblastoma cells with bradykinin induces phosphorylation of threonine residues in the myosin-II heavy chain; this phosphorylation is Ca2+ dependent and regulated by Rac. Both bradykinin-mediated and constitutive activation of Rac promote cell spreading, accompanied by a loss of cortical myosin II. Our results identify the myosin-II heavy chain as a new target of Rac-regulated kinase pathways, and implicate Rac as a Rho antagonist during myosin-II-dependent cell-shape changes.

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Figure 1: Bradykinin activates Rac and induces phosphorylation of the myosin-II heavy chain.
Figure 2: MHC phosphorylation is regulated by Rac activity.
Figure 3: Bradykinin and activation of Rac induce cell spreading and redistribution of myosin II in N1E-115 cells; constitutively active RhoAV14 and dominant-negative Pak inhibit spreading.
Figure 4: Involvement of Pak kinases and Ca2+ in MHC phosphorylation.
Figure 5: Signalling pathways regulating neuroblastoma morphology.

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Acknowledgements

We thank A. Hall for providing Rac and Rho cDNAs; G. Nolan for providing phoenix eco- and amphotrophic packaging cells and LZRS retroviral vectors; and L.Oomen and N. Ong for help in making the figures. This work was supported by grants from the Dutch Cancer Society to J.G.C.

Correspondence and requests for materials should be addressed to J.G.C.

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  1. Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, Amsterdam, 1066 CX, The Netherlands

    Frank N. van Leeuwen, Sanne van Delft, Hendrie E. Kain, Rob A. van der Kammen & John G. Collard

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Correspondence to John G. Collard.

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van Leeuwen, F., van Delft, S., Kain, H. et al. Rac regulates phosphorylation of the myosin-II heavy chain, actinomyosin disassembly and cell spreading. Nat Cell Biol 1, 242–248 (1999). https://doi.org/10.1038/12068

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