Critical determinants of the G protein γ subunits in the Gβγ stimulation of G protein-activated inwardly rectifying potassium (GIRK) channel activity

The betagamma subunits of G proteins modulate inwardly rectifying potassium (GIRK) channels through direct interactions. Although GIRK currents are stimulated by mammalian Gbetagamma subunits, we show that they were inhibited by the yeast Gbetagamma (Ste4/Ste18) subunits. A chimera between the yeast and the mammalian Gbeta(1) subunits (ymbeta) stimulated or inhibited GIRK currents, depending on whether it was co-expressed with mammalian or yeast Ggamma subunits, respectively. This result underscores the critical functional influence of the Ggamma subunits on the effectiveness of the Gbetagamma complex. A series of chimeras between Ggamma(2) and the yeast Ggamma revealed that the C-terminal half of the Ggamma(2) subunit is required for channel activation by the Gbeta(gamma) complex. Point mutations of Ggamma(2) to the corresponding yeast Ggamma residues identified several amino acids that reduced significantly the ability of Gbeta(gamma) to stimulate channel activity, an effect that was not due to improper association with Gbeta. Most of the identified critical Ggamma residues clustered together, forming an intricate network of interactions with the Gbeta subunit, defining an interaction surface of the Gbetagamma complex with GIRK channels. These results show for the first time a functional role for Ggamma in the effector role of Gbetagamma.