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Autoinhibition and activation mechanisms of the Wiskott–Aldrich syndrome protein


The Rho-family GTPase, Cdc42, can regulate the actin cytoskeleton through activation of Wiskott–Aldrich syndrome protein (WASP) family members. Activation relieves an autoinhibitory contact between the GTPase-binding domain and the carboxy-terminal region of WASP proteins. Here we report the autoinhibited structure of the GTPase-binding domain of WASP, which can be induced by the C-terminal region or by organic co-solvents. In the autoinhibited complex, intramolecular interactions with the GTPase-binding domain occlude residues of the C terminus that regulate the Arp2/3 actin-nucleating complex. Binding of Cdc42 to the GTPase-binding domain causes a dramatic conformational change, resulting in disruption of the hydrophobic core and release of the C terminus, enabling its interaction with the actin regulatory machinery. These data show that ‘intrinsically unstructured’ peptides such as the GTPase-binding domain of WASP can be induced into distinct structural and functional states depending on context.

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Figure 1: Amino-acid sequences of human WASP and N-WASP with consensus secondary structure of the GBD constructs of WASP shown above.
Figure 2: Biochemical characterization of the WASP GBD.
Figure 3: Stereoviews of the best-fit superpositions of the backbone (N, Ca, C) atoms of the 20 final NMR structures of GBD2 in 5% TFE (a) and GBD4–C (b).
Figure 4: Autoinhibited and activated structures of the WASP GBD.
Figure 5: Steric incompatibility of the autoinhibited and activated structures.


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We thank M. Lu for analytical ultracentrifugation experiments; W. Hu for communicating results before publication; M. Buck, Y.M. Chook, J. Goldberg, B. Schulman, B. W. Trotter and M. R. Wood for discussion and critical reading of the manuscript; R. Cerione and K. Siminovitch for providing cDNAs for Cdc42 and WASP, respectively; L. Kay for providing many of the NMR pulse sequences used in this study; F. Delaglio for providing nmrPipe and TALOS software; A. Majumdar, W. Hu and B. Aghazadeh for assistance in NMR data acquisition; J. Hubbard for computer system support and M. Fiore for expert administrative assistance. A.S.K. is supported by a fellowship from the Damon Runyon-Walter Winchell Foundation, and L.T.K. by the Charles H. Revson Foundation. M.K.R. acknowledges support from the NIH (PECASE program), Arnold and Mabel Beckman Foundation, and Sidney Kimmel Foundation for Cancer Research.

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Correspondence to Michael K. Rosen.

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Kim, A., Kakalis, L., Abdul-Manan, N. et al. Autoinhibition and activation mechanisms of the Wiskott–Aldrich syndrome protein. Nature 404, 151–158 (2000).

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