Heterotrimeric G Protein Subunit Gαq Is a Master Switch for Gβγ-Mediated Calcium Mobilization by Gi-Coupled GPCRs

Abstract

Mechanisms that control mobilization of cytosolic calcium [Ca²⁺]_i are key for regulation of numerous eukaryotic cell functions. One such paradigmatic mechanism involves activation of phospholipase Cβ (PLCβ) enzymes by G protein βγ subunits from activated Gα_i-Gβγ heterotrimers. Here, we report identification of a master switch to enable this control for PLCβ enzymes in living cells. We find that the Gα_i-Gβγ-PLCβ-Ca²⁺ signaling module is entirely dependent on the presence of active Gα_q. If Gα_q is pharmacologically inhibited or genetically ablated, Gβγ can bind to PLCβ but does not elicit Ca²⁺ signals. Removal of an auto-inhibitory linker that occludes the active site of the enzyme is required and sufficient to empower "stand-alone control" of PLCβ by Gβγ. This dependence of Gi-Gβγ-Ca²⁺ on Gα_q places an entire signaling branch of G-protein-coupled receptors (GPCRs) under hierarchical control of Gq and changes our understanding of how Gi-GPCRs trigger [Ca²⁺]_i via PLCβ enzymes.

Keywords: Ca(2+) signaling; FR900359; GPCR; GPR17; Gi; Gq; PTX; heterotrimeric G protein; phospholipase Cβ; real-time BRET-based IP(3) biosensor.