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- _5 value "We also employed double-strand (ds) RNA mediated interference (RNAi) (Elbashir et al., 2001) to deplete endogenous Daam1 protein. The ds RNAi oligo for Daam1 reduces the protein level of Daam1, but not of RhoA, ?-catenin (Figure 3C) or Dvl2 (not shown). Importantly, the RNAi oligo inhibits RhoA activation by Wnt-1, Fz, or Dvl, but not by Ephexin (Figure 3C). A control ds RNAi oligo has no effect on Daam1 or RhoA protein level or RhoA activation (Figure 3C). Thus, Daam1 is essential for Wnt/Fz/Dvl activation of RhoA. AN ACTIVATED DAAM1 INDUCES RHO ACTIVATION IN A RHO-GEF-DEPENDENT MANNER Previous studies suggest that deletion of the amino terminal domain results in constitutive activation of FH proteins by releasing an intramolecular inhibition (Watanabe et al. 1999; Tominaga et al. 2000 and Westendorf 2001). C-Daam1 thus may represent an activated form of Daam1. Indeed, while the full length Daam1 is inactive, C-Daam1 overexpression causes activation of RhoA, but not of Rac or Cdc42 (Figure 3D). C-Daam1 activation of RhoA is not affected by ?PDZ-Dvl (Figure 3A), which inhibits RhoA activation by Fz or Dvl (Figure 1F), further demonstrating that Daam1 functions downstream of Dvl in Rho activation." provenance.
- _5 wasQuotedFrom 11779461 provenance.