The Rho GEFs LARG and GEF-H1 regulate the mechanical response to force on integrins

Abstract

How individual cells respond to mechanical forces is of considerable interest to biologists as force affects many aspects of cell behaviour1. The application of force on integrins triggers cytoskeletal rearrangements and growth of the associated adhesion complex, resulting in increased cellular stiffness2,3, also known as reinforcement4. Although RhoA has been shown to play a role during reinforcement3, the molecular mechanisms that regulate its activity are unknown. By combining biochemical and biophysical approaches, we identified two guanine nucleotide exchange factors (GEFs), LARG and GEF-H1, as key molecules that regulate the cellular adaptation to force. We show that stimulation of integrins with tensional force triggers activation of these two GEFs and their recruitment to adhesion complexes. Surprisingly, activation of LARG and GEF-H1 involves distinct signalling pathways. Our results reveal that LARG is activated by the Src family tyrosine kinase Fyn, whereas GEF-H1 catalytic activity is enhanced by ERK downstream of a signalling cascade that includes FAK and Ras.

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Figure 1: LARG and GEF-H1 activate RhoA in response to force.
Figure 2: LARG and GEF-H1 mediate cellular stiffening in response to force applied on integrins.
Figure 3: Fyn mediates LARG activation in response to force.
Figure 4: ERK activates GEF-H1 in response to force.

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Acknowledgements

The authors would like to thank L. Sharek for her technical support. This study was supported by National Institutes of Health Grant nos GM029860 and GM029860-28S (to K.B.), P41-EB002025-23A1 (R.S.) and R01-HL077546-03A2 (R.S.), and a grant from the University Cancer Research Fund from the Lineberger Comprehensive Cancer Center. C.G. is supported by a Marie Curie Outgoing International Fellowship from the European Union Seventh Framework Programme (FP7/2007–2013) under grant agreement no. 254747.

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C.G. and V.S. designed and carried out experiments. R.G.M. and E.T.O. helped with experimental design and procedures. C.G. and K.B. wrote the manuscript. K.B. and R.S. directed the project and revised the manuscript. All authors provided detailed comments.

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Correspondence to Keith Burridge.

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Guilluy, C., Swaminathan, V., Garcia-Mata, R. et al. The Rho GEFs LARG and GEF-H1 regulate the mechanical response to force on integrins. Nat Cell Biol 13, 722–727 (2011). https://doi.org/10.1038/ncb2254

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