DDX proteins directly bind to protein kinases and can stimulate their enzymatic activity. (A) Outline of the protein ProtoArray binding screen. (B) Examples of DDX-interacting kinases on protein ProtoArray interrogated with V5-tagged recombinant DDX or GFP proteins. (C) Protein domain enrichment analysis (Pfam terms) of ProtoArray proteins bound specifically to DDX39A or DDX56. (D) Overlap between protein kinase hits from the DDX interactome and ProtoArray binding screen. (E) Outline of the kinome-wide activity screen. (F) Ranked results from an in vitro kinase activity screen with the indicated DDX proteins. Every point corresponds to one tested kinase. Kinases stimulated by a DDX protein with log2 fold change above 0.5 (red line) were considered positive hits. (G) Overview of all kinases identified to be stimulated by any of the four DDX proteins. (H) Venn diagram of all kinases identified in the ProtoArray and kinase activity screen showing an overlap of 16 common kinase hits. (I) Venn diagram of all kinase hits from all three screening approaches, showing six kinases being identified to be bound and regulated by DDX proteins in three independent screens.
