Myosin VI and B-arrestin synergistically regulate GIPR internalization and signaling

Glucose-dependent insulinotropic peptide receptor (GIPR) stimulates insulin release and regulates metabolic homeostasis. GIPR function is shaped by spatiotemporal trafficking of this G protein-coupled receptor (GPCR). While GPCR endocytosis is traditionally associated with β-arrestin, GIPR internalization is only modestly dependent on this pathway. In this study, we demonstrate that GIPR engages a cytoskeletal motor, myosin VI to drive receptor endocytosis. GIPR engages the adaptor-motor complex through a PDZ-binding motif (PBM) at its C-ail. Interestingly, β-arrestin binding to phosphorylated residues upstream of the PBM enhance myosin VI recruitment and activation. GIPR internalization is dependent on both receptor phosphorylation and the PBM site to recruit β-arrestin and myosin VI, respectively. Synergistic engagement of β-arrestin and myosin VI results in desensitization of GIP-stimulated cAMP signaling while activating pERK1/2 from endosomal compartments. Blocking myosin VI activity enhances insulin release in pancreatic beta cells, demonstrating a novel role for this pathway in regulating the physiological effects of GIPR. Our findings highlight the direct convergence of two independent trafficking pathways at the level of the receptor C-tail, with implications for the nuanced regulation of individual GPCRs through the differential engagement of β-arrestin and myosin VI.