Fig. 3.

Class A GPCR and G-protein mutually exclusive mutations: a as for Fig. 2a, but for GPCR and Gα mutations at either GPCR (DRY) R3.50 or Gα SWI arginine. Only the top 30 mutated genes are shown. b GPCR (PDB: 3NYA) and c Gα (1AZT) significant positions indicated as spheres centred on Cα atoms and whose diameter is proportional to the number of mutations. The right panel shows GPCRs coupling preferences from IUPHAR (maroon and red indicate primary and secondary coupling respectively). The lower panel shows co-occurring mutations for the top 10 most mutated signalling oncodrivers; d Loss of G-protein signalling activity in the DRY mutant GPCRs. HEK293 cells transfected with the alkaline phosphatase-tagged transforming growth factor-α (AP-TGFα)-encoding plasmid together with an empty plasmid (Mock), WT GPCR-encoding plasmid (WT) or DRY-mutant GPCR-encoding plasmid (MT) treated with titrated ligands for 1 h while quantifiying AP-TGFα release into conditioned media. Symbols and error bars represent mean and SEM, respectively, of three to five independent experiments with each measured in triplicates. For MRGPRX1 and HCRTR2, symbols of MT overlap with Mock. Parameters from the concentration-response curves (EC₅₀ and Emax) are listed in Table S15; e Kaplan-Meier curve showing survival analysis for patients affected by R^3.50 mutations (orange curves) in skin melanoma