Switching Heterotrimeric G Protein Subunits with a Chemical Dimerizer

SummaryThe selective manipulation of single intracellular-signaling events remains one of the key tasks when studying signaling networks. Here, we demonstrate for the first time the stimulation of FKBP fusions of various subunits of heterotrimeric G proteins by the simple addition of the chemical dimerizer rapamycin. Activation of constitutively active Gαq, but not its GDP-bound form, leads to sustained oscillations of intracellular calcium and myo-inositol 1,4,5-trisphosphate (InsP3) levels in HEK cells, independent of the activation of endogenous Gαq, in full agreement with the InsP3-Ca2+ cross-coupling model of calcium oscillations. Rapamycin-induced translocation of wild-type Gαs to the plasma membrane results in elevated cAMP levels. Activation of rapamycin-inducible Gαs or Gβ1γ2 evokes extensive modulation of ATP-induced calcium transients. The results demonstrate that inducible heterotrimeric G protein subunits will provide ways for dissecting G protein-coupled receptor signaling.

► Rapid and selective activation of single G protein subunits via a chemical dimerizer
► Inducible Gαq generates prolonged, highly regular calcium oscillations, indicating that the method likely triggers the basic oscillatory machinery
► Each cell produces a unique calcium response pattern, independent of the Gαq-FKBP expression level
► Inducible Gαs triggers sustained cAMP production
► Activation of inducible Gαs and Gβ1γ2 modulates cellular calcium transients