Abstract
Individual tumour cells move in three-dimensional environments with either a rounded or an elongated ‘mesenchymal’ morphology. These two modes of movement are tightly regulated by Rho family GTPases: elongated movement requires activation of Rac1, whereas rounded/amoeboid movement engages specific Cdc42 and Rho signalling pathways. In siRNA screens targeting the genes encoding guanine nucleotide exchange factors (GEFs), we found that the Ras GEF RasGRF2 regulates conversion between elongated- and rounded-type movement. RasGRF2 suppresses rounded movement by inhibiting the activation of Cdc42 independently of its capacity to activate Ras. RasGRF2 and RasGRF1 directly bind to Cdc42, outcompeting Cdc42 GEFs, thereby preventing Cdc42 activation. By this mechanism, RasGRFs regulate other Cdc42-mediated cellular processes such as the formation of actin spikes, transformation and invasion in vitro and in vivo. These results demonstrate a role for RasGRF GEFs as negative regulators of Cdc42 activation.
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Change history
15 July 2011
In the version of this article initially published online and in print, the panel on the lower right-hand side of Fig. 3a corresponding to the anti-SOS blot was incorrectly duplicated and shown in place of the anti-Cdc42 blot.
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Acknowledgements
We are grateful to M. Moran, L. Feig and X. Bustelo for providing reagents. P.C.’s laboratory is supported by grants BFU2008-01728 from the Spanish Ministry of Education, GROWTHSTOP (LSHC CT-2006-037731) from the EU VI Framework Program and Red Temática de Investigación Cooperativa en Cáncer (RTICC; RD06/0020/0105), Spanish Ministry of Health. C.J.M.’s laboratory is supported by Cancer Research UK. C.J.M. is a Gibb Life Fellow of Cancer Research UK.
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F.C. carried out all of the experiments with the exception of those shown in Figs 1 and 6, which were carried out by F.C. and V.S-M. V.S-M. and F.W. carried out the invasion assays and imaging of the invading cells. L.A-I. carried out the in vitro GTPase binding experiments and the soft-agar colonies. F.C. and V.S-M. also prepared the figures and carried out the statistical analyses. E.S. contributed in the lung colonization assays. F.C., V.S-M., C.J.M. and P.C. conceived the study and C.J.M. and P.C. wrote the manuscript.
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Calvo, F., Sanz-Moreno, V., Agudo-Ibáñez, L. et al. RasGRF suppresses Cdc42-mediated tumour cell movement, cytoskeletal dynamics and transformation. Nat Cell Biol 13, 819–826 (2011). https://doi.org/10.1038/ncb2271
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DOI: https://doi.org/10.1038/ncb2271
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