Gαq directly activates p63RhoGEF and trio via a conserved extension of the Dbl homology-associated pleckstrin homology domain

The coordinated cross-talk from heterotrimeric G proteins to Rho GTPases is essential during a variety of physiological processes. Emerging data suggest that members of the G alpha(12/13) and G alpha(q/11) families of heterotrimeric G proteins signal downstream to RhoA via distinct pathways. Although studies have elucidated mechanisms governing G alpha(12/13)-mediated RhoA activation, proteins that functionally couple G alpha(q/11) to RhoA activation have remained elusive. Recently, the Dbl- family guanine nucleotide exchange factor (GEF) p63RhoGEF/GEFT has been described as a novel mediator of G alpha(q/11) signaling to RhoA based on its ability to synergize with G alpha(q/11) resulting in enhanced RhoA signaling in cells. Wehave used biochemical/biophysical approaches with purified protein components to better understand the mechanism by which activated G alpha(q) directly engages and stimulates p63RhoGEF. Basally, p63RhoGEF is autoinhibited by the Dbl homology (DH)-associated pleckstrin homology (PH) domain; activated G alpha(q) relieves this autoinhibition by interacting with a highly conserved C-terminal extension of the PH domain. This unique extension is conserved in the related Dbl- family members Trio and Kalirin and we show that the C-terminal Rho-specific DH-PH cassette of Trio is similarly activated by G alpha(q)