Abstract
The Rho-family GTP-hydrolysing proteins (GTPases), Cdc42, Rac and Rho, act as molecular switches in signalling pathways that regulate cytoskeletal architecture, gene expression and progression of the cell cycle1. Cdc42 and Rac transmit many signals through GTP-dependent binding to effector proteins containing a Cdc42/Rac-interactive-binding (CRIB) motif2. One such effector, the Wiskott–Aldrich syndrome protein (WASP), is postulated to link activation of Cdc42 directly to the rearrangement of actin3. Human mutations in WASP cause severe defects in haematopoletic cell function, leading to clinical symptoms of thrombocytopenia, immunodeficiency and eczema. Here we report the solution structure of a complex between activated Cdc42 and a minimal GTPase-binding domain (GBD) from WASP. An extended amino-terminal GBD peptide that includes the CRIB motif contacts the switch I, β2 and α5 regions of Cdc42. A carboxy-terminal β-hairpin and α-helix pack against switch II. The Phe-X-His-X2-His portion of the CRIB motif and the α-helix appear to mediate sensitivity to the nucleotide switch through contacts to residues 36–40 of Cdc42. Discrimination between the Rho-family members is likely to be governed by GBD contacts to the switch I and α5 regions of the GTPases. Structural and biochemical data suggest that GBD-sequence divergence outside the CRIB motif may reflect additional regulatory interactions with functional domains that are specific to individual effectors.
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Acknowledgements
We thank Y.M. Chook, A. Kim and J. Goldberg for discussion and for critically reading the manuscript; R. Cerione for Cdc42 cDNA; R. Goody for initial sample of GMPPCP and for advice on synthesis of the nucleotide; L. Kay for many of the NMR pulse sequences; F. Delaglio for unpublished TALOS software; I. Armitage and D. Live for assistance with data collection at the Structural Biology NMR Resource at the University of Minnesota Medical School; J. Hubbard for computer-system support; and S. Freihaut for administrative assistance. B.A. is supported by a grant from the US Army Breast Cancer program. M.K.R. acknowledges support from the NIH (PECASE program), the Arnold and Mabel Beckman Foundation, and the Sidney Kimmel Foundation for Cancer Research.
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Abdul-Manan, N., Aghazadeh, B., Liu, G. et al. Structure of Cdc42 in complex with the GTPase-binding domain of the ‘Wiskott–Aldrich syndrome’ protein. Nature 399, 379–383 (1999). https://doi.org/10.1038/20726
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DOI: https://doi.org/10.1038/20726
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