Abstract
Members of the Wiskott-Aldrich syndrome protein (WASP) family link Rho GTPase signaling pathways to the cytoskeleton through a multiprotein assembly called Arp2/3 complex. The C-terminal VCA regions (verprolin-homology, central hydrophobic, and acidic regions) of WASP and its relatives stimulate Arp2/3 complex to nucleate actin filament branches. Here we show by differential line broadening in NMR spectra that the C (central) and A (acidic) segments of VCA domains from WASP, N-WASP and Scar bind Arp2/3 complex. The C regions of these proteins have a conserved sequence motif consisting of hydrophobic residues and an arginine residue. Point mutations in this conserved sequence motif suggest that it forms an amphipathic helix that is required in biochemcial assays for activation of Arp2/3 complex. Key residues in this motif are buried through contacts with the GTPase binding domain in the autoinhibited structure of WASP and N-WASP, indicating that sequestration of these residues is an important aspect of autoinhibition.
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Acknowledgements
We thank S. Padrick for writing Mathematica scripts used in the analysis of anisotropy data. This work was supported by grants from the US National Institutes of Health (NIH) and Welch Foundation to M.K.R. and an NIH research grant to T.D.P.
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Panchal, S., Kaiser, D., Torres, E. et al. A conserved amphipathic helix in WASP/Scar proteins is essential for activation of Arp2/3 complex. Nat Struct Mol Biol 10, 591–598 (2003). https://doi.org/10.1038/nsb952
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DOI: https://doi.org/10.1038/nsb952
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