RPS15 mutant proteins alter global cell proteome in HEK293T cells. (A) Heat map depicting unsupervised hierarchical clustering of the 8 proteomes analyzed using the total set of proteins identified. Protein abundance is shown as the log2 ratio between each sample with respect to the average of reference samples (GFP-RPS15^WT–expressing HEK293T cells). (B) Principal component analysis of the relative protein abundances of all 8 proteomes analyzed. Principal component analyses show a similar clustering between biological replicates. (C-D) Comparison of proteomic profiles from GFP-RPS15^P131S–expressing vs GFP-RPS15^WT–expressing HEK293T cells performed using GSEA. (C) Enrichment score plot corresponding to DNA strand elongation in the Reactome database (left panel). Heat map showing the top 18 genes of the positively correlated enriched gene set (right panel). (D) Enrichment plot corresponding to activation of the mRNA upon binding of the CAP-binding complex and eukaryotic initiation factors (eIFs) in the Reactome database (left panel). Heat map showing the 19 top genes of the negatively correlated enriched gene set (right panel). (E) Comparison of proteomic profiles from GFP-RPS15^S138F–expressing vs GFP-RPS15^WT–expressing HEK293T cells performed using GSEA. The enrichment plot shown corresponds to destabilization of mRNA by KSRP in the Reactome database (left panel). Heat map showing the top 16 genes of the positively correlated enriched gene set (right panel). (F) Relative abundance of different components of the exosome complex. Each bar represents the mean of 2 (GFP-RPS15^WT and GFP-RPS15^P131S) or 4 (GFP-RPS15^S138F) independent relative proteomic determinations + SEM. *P < .033, ***P < .001, multiple t test.