Fig. 4.

Functional consequence of GPCR mutations: a integrative analysis of G-protein activities, hotspot mutations and GPCR deleterious mutations in different TCGA cancer types. Top panel: G-protein activities estimated by combining G-proteins and receptors differential expression levels through coupling information; middle panel: fraction of samples with deleterious GPCR mutations (i.e., either highly conserved residues, stop gains or frameshifts); lower panel: known activating GNAS (p.R201) and GNAQ (p.Q209) mutations. b Median of RPKM values of Gα subunits in 32 TCGA cancer types. c Number of unique samples (crimson) and fraction of the coupling group (grey) displaying deleterious mutations pan-cancer; d Loss of G_i activity in the DRY mutant GPCRs. HEK293 cells transfected with the cAMP biosensor-encoding plasmid and plasmid together with an empty plasmid (Mock), WT GPCR-encoding plasmid (WT) or DRY-mutant GPCR-encoding plasmid (MT) were loaded with D-luciferin for 2 h and treated with titrated ligands in the presence of 10 µM forskolin for 10 min. Luminescent signals were measured before and after ligand addition and data expressed as a change in luminescent counts. Symbols and error bars represent mean and SEM, respectively, of six to seven independent experiments with each measured in duplicates; e cartoons of the Adenylate Cyclase pathway regulation summarising the mechanisms described in the text