Fig. 3.

Wash preferentially binds active Rho but not Rac or Cdc42. (A) Summary of the previously known associations among Rho GTPases, WASP family proteins and actin nucleators. (B) GST pull-down assays with ³⁵S-labeled in vitro translated proteins on the left and GST fusion proteins or GST alone across the top. Wash binds to Rho1^GTP, but not Rac or Cdc42. By comparison, Wasp binds Cdc42^GTP, whereas Scar binds directly to none of the three effectors. A fourth WASP subfamily protein in Drosophila, Whamy (CG12946), binds Cdc42^GTP. (C) GST pull-down assay with purified His-Wash and GST-Rho family proteins. Wash binds directly to Rho1^GTP, but not Rac or Cdc42. All assays were performed a minimum of three times.