Disruption of the interaction between mutationally activated Gα_q and Gβγ attenuates aberrant signaling

Heterotrimeric G protein stimulation via G protein-coupled receptors promotes downstream proliferative signaling. Mutations can occur in Gα proteins which prevent GTP hydrolysis; this allows the G proteins to signal independently of G protein-coupled receptors and can result in various cancers, such as uveal melanoma (UM). Most UM cases harbor Q209L, Q209P, or R183C mutations in Gα_q/11 proteins, rendering the proteins constitutively active (CA). Although it is generally thought that active, GTP-bound Gα subunits are dissociated from and signal independently of Gβγ, accumulating evidence indicates that some CA Gα mutants, such as Gα_q/11, retain binding to Gβγ, and this interaction is necessary for signaling. Here, we demonstrate that disrupting the interaction between Gβγ and Gα_q is sufficient to inhibit aberrant signaling driven by CA Gα_q. Introduction of the I25A point mutation in the N-terminal α helical domain of CA Gα_q to inhibit Gβγ binding, overexpression of the G protein Gα_o to sequester Gβγ, and siRNA depletion of Gβ subunits inhibited or abolished CA Gα_q signaling to the MAPK and YAP pathways. Moreover, in HEK 293 cells and in UM cell lines, we show that Gα_q-Q209P and Gα_q-R183C are more sensitive to the loss of Gβγ interaction than Gα_q-Q209L. Our study challenges the idea that CA Gα_q/11 signals independently of Gβγ and demonstrates differential sensitivity between the Gα_q-Q209L, Gα_q-Q209P, and Gα_q-R183C mutants.