A regulator of G protein signaling interaction surface linked to effector specificity

Proteins of the regulator of G protein signaling (RCS) family accelerate GT P hydrolysis by the or subunits (G(alpha)) of G proteins. leading to rapid recovery of signaling cascades. Many different RCS proteins can accelerate GTP hydrolysis by an individual G(alpha), and CTP hydrolysis rates of diffe rent G alpha(s) can be enhanced by the same RCS protein. Consequently, the mechanisms for specificity in RGS regulation and the residues involved rema in unclear. Using the evolutionary trace (ET) method, we have identified a cluster of residues in the RGS domain that includes the RCS-C, binding inte rface and extends to include additional functionally important residues on the surface. One of these is within helix alpha 3, two are in alpha 5, and three are in the loop connecting alpha 5 and alpha 6. A cluster of surface residues on G(alpha) previously identified by ET, and composed predominantl y of residues from the switch III region and helix alpha 3, is spatially co ntiguous with the PT-identified residues in the RGS domain. This cluster in cludes residues proposed to interact with the gamma subunit of G(t alpha)'s effector, cGMP phosphodiesterase (PDE gamma), The proximity of these clust ers suggests that they form part of an interface between the effector and t he RGS-G(alpha) complex. Sequence variations in these residues correlate wi th PDE gamma effects on GTPase acceleration. Because ET identifies residues important for all members of a protein family, these residues likely form a general site for regulation of G protein-coupled signaling cascades, poss ibly by means of effector interactions.