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A novel and evolutionarily conserved PtdIns(3,4,5)P3-binding domain is necessary for DOCK180 signalling

Nature Cell Biology volume 7, pages 797–807 (2005)Cite this article

Abstract

The evolutionarily conserved DOCK180 protein has an indispensable role in cell migration by functioning as an exchange factor for Rac GTPase via its DOCK homology region (DHR)-2 domain. We report here that the conserved DHR-1 domain also has an important signalling role. A form of DOCK180 that lacks DHR-1 fails to promote cell migration, although it is capable of inducing Rac GTP-loading. The DHR-1 domain interacts with PtdIns(3,4,5)P3 in vitro and in vivo, and mediates the DOCK180 signalling complex localization at sites of PtdIns(3,4,5)P3 accumulation in the cell's leading edge. A form of DOCK180 in which the DHR-1 domain has been replaced by a canonical PtdIns(3,4,5)P3-binding pleckstrin homology domain is fully functional at inducing cell elongation and migration, suggesting that the main function of DHR-1 is to bind PtdIns(3,4,5)P3. These results demonstrate that DOCK180, via its DHR-1 and DHR-2 domains, couples PtdIns(3,4,5)P3 signalling to Rac GTP-loading, which is essential for directional cell movement.

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Figure 1: The DHR-1 domain is required for DOCK180-mediated cell elongation and motility.
Figure 2: The DHR-1 domain is required for cell spreading.
Figure 3: Intact GTP-loading of Rac and coupling to ELMO1 and CrkII by the DHR-1 mutant of DOCK180.
Figure 4: The DHR-1 domain of DOCK180 has lipid-binding activity towards phosphoinositides in vitro.
Figure 5: The DHR-1 domain shows lipid-binding activity in vivo.
Figure 6: Generation of PtdIns(3,4,5)P3 is necessary for DOCK180-induced cell elongation and migration.
Figure 7: The PH domain of BMX/Etk can functionally replace the DHR-1 domain in DOCK180.

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Acknowledgements

We thank M. Hurwitz, H. R. Horvitz, D. Schlaepfer and H. Katoh for useful discussions. We acknowledge the expert technical help provided by E. Monosov for microscopy experiments. J.-F.C. is a Research Fellow of The Terry Fox Foundation/National Cancer Institute of Canada. This work was supported by grants from the National Institutes of Health (to K.V.).

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  1. Jean-François Côté

    Present address: Clinical Research Institute of Montreal, 110 Pine Avenue West, Montreal, Quebec, Canada, H2W 1R7

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  1. The Burnham Institute, 10901 North Torrey Pines Road, La Jolla, 92037, CA, USA

    Jean-François Côté, Andrea B. Motoyama, Jason A. Bush & Kristiina Vuori

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Côté, JF., Motoyama, A., Bush, J. et al. A novel and evolutionarily conserved PtdIns(3,4,5)P3-binding domain is necessary for DOCK180 signalling. Nat Cell Biol 7, 797–807 (2005). https://doi.org/10.1038/ncb1280

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