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The Rac1- and RhoG-specific GEF domain of Trio targets filamin to remodel cytoskeletal actin

Nature Cell Biology volume 2, pages 888–892 (2000)Cite this article

Abstract

Rho GTPases control actin reorganization and many other cellular functions. Guanine nucleotide-exchange factors (GEFs) activate Rho GTPases by promoting their exchange of GDP for GTP. Trio is a unique Rho GEF, because it has separate GEF domains, GEFD1 and GEFD2, that control the GTPases RhoG/Rac1 and RhoA, respectively. Dbl-homology (DH) domains that are common to GEFs catalyse nucleotide exchange, and pleckstrin-homology (PH) domains localize Rho GEFs near their downstream targets. Here we show that Trio GEFD1 interacts through its PH domain with the actin-filament-crosslinking protein filamin, and localizes with endogenous filamin in HeLa cells. Trio GEFD1 induces actin-based ruffling in filamin-expressing, but not filamin-deficient, cells and in cells transfected with a filamin construct that lacks the Trio-binding domain. In addition, Trio GEFD1 exchange activity is not affected by filamin binding. Our results indicate that filamin, as a molecular target of Trio, may be a scaffold for the spatial organization of Rho-GTPase-mediated signalling pathways.

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Figure 1: Trio GEFD1 interacts specifically with filamin A.
Figure 2: Trio GEFD1 binds to filamin in vitro.
Figure 3: Localization of Trio and filamin in Trio GEFD1-induced membrane ruffles.
Figure 4: Filamin A expression is required for Trio GEFD1-induced actin-cytoskeleton modification.
Figure 5: The capacity of Trio GEFD1 to remodel the actin cytoskeleton is directly linked to its binding to filamin.
Figure 6: Filamin is dispensable for Trio GEFD1-elicited JNK stimulation.
Figure 7: Filamin does not affect the GDP/GTP exchange activity of Trio GEFD1.

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Acknowledgements

We thank S. Schmidt and S. Estrach for discussions, S. Diriong for technical assistance and P. Travo, Head of the CRBM Integrated Imaging Facility, for interest and support. Confocal analysis was carried out at the Centre Regional d'Imagerie Cellulaire, Montpellier, with the help of N. Lautredou. This work was funded by CNRS institutional grants, contracts from the Ligue Nationale contre le Cancer, the Association pour la Recherche contre le Cancer, and USPHS NIH grant HL19429.

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Authors and Affiliations

  1. CRBM-CNRS, UPR 1086, 1919 Route de Mende , Montpellier, 34293, Cédex 5, France

    Jean-Michel Bellanger & Anne Debant

  2. LMC, UMR 5539, USTL, Montpellier , 34090, France

    Catherine Astier

  3. IGMM, UMR 5535, Montpellier , 34293, France

    Claude Sardet

  4. Hematology Division, Brigham and Women's Hospital and Department of Medicine, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Yasutaka Ohta & Thomas P. Stossel

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Correspondence to Anne Debant.

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Bellanger, JM., Astier, C., Sardet, C. et al. The Rac1- and RhoG-specific GEF domain of Trio targets filamin to remodel cytoskeletal actin. Nat Cell Biol 2, 888–892 (2000). https://doi.org/10.1038/35046533

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