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Structural basis for the selective activation of Rho GTPases by Dbl exchange factors

Nature Structural Biology volume 9, pages 468–475 (2002)Cite this article

Abstract

Activation of Rho-family GTPases involves the removal of bound GDP and the subsequent loading of GTP, all catalyzed by guanine nucleotide exchange factors (GEFs) of the Dbl-family. Despite high sequence conservation among Rho GTPases, Dbl proteins possess a wide spectrum of discriminatory potentials for Rho-family members. To rationalize this specificity, we have determined crystal structures of the conserved, catalytic fragments (Dbl and pleckstrin homology domains) of the exchange factors intersectin and Dbs in complex with their cognate GTPases, Cdc42 and RhoA, respectively. Structure-based mutagenesis of intersectin and Dbs reveals the key determinants responsible for promoting exchange activity in Cdc42, Rac1 and RhoA. These findings provide critical insight into the structural features necessary for the proper pairing of Dbl-exchange factors with Rho GTPases and now allow for the detailed manipulation of signaling pathways mediated by these oncoproteins in vivo.

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Figure 1: Overall structures of ITSN–Cdc42 and Dbs–RhoA.
Figure 2: Residues in Rho GTPases and Rho–GEFs that dictate specificity.
Figure 3: Structural elements of ITSN that prevent nucleotide exchange in Rac1 and RhoA.
Figure 4: Specific substitutions in exchange factors alter selectivity for Rho GTPase substrates.
Figure 5: Comparison of the Dbs–RhoA, Dbs–Cdc42 and Tiam1–Rac1 structures.

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Acknowledgements

We thank M. Kimple, C. Ogata and the staff of X4a at the Brookhaven National Light Source and F. Rotello and the staff of BM19 at the Advanced Photon Source for assistance with data collection. D.K.W. is supported by a grant from the American Cancer Society, and J.S. acknowledges support from the NIH and the Pew Charitable Trusts.

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Author notes

  1. Jason T. Snyder, David K. Worthylake, Kent L. Rossman, Laurie Betts, Wendy M. Pruitt, David P. Siderovski, Channing J. Der and John Sondek: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Biophysics, Program in Molecular and Cellular Biophysics, Chapel Hill, 27599, North Carolina, USA

    Jason T. Snyder, Kent L. Rossman & John Sondek

  2. Department of Pharmacology, University of North Carolina at Chapel Hill, Lineberger Comprehensive Cancer Center, Chapel Hill, 27599, North Carolina, USA

    David K. Worthylake, Laurie Betts, Wendy M. Pruitt, David P. Siderovski, Channing J. Der & John Sondek

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Correspondence to John Sondek.

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Snyder, J., Worthylake, D., Rossman, K. et al. Structural basis for the selective activation of Rho GTPases by Dbl exchange factors. Nat Struct Mol Biol 9, 468–475 (2002). https://doi.org/10.1038/nsb796

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