Phosphorylation-mediated regulation of integrin-linked kinase 5 by purinoreceptor P2K2.

Extracellular ATP (eATP) in plants plays a crucial role as a ligand for purinoreceptors, mediating purinergic signaling and regulating diverse biological functions, including responses to abiotic and biotic stresses. DORN1/P2K1 (LecRK I.9) was the first identified plant purinoreceptor. P2K2 (LecRK I.5) was subsequently identified as an additional plant purinoreceptor and shown to directly interact with P2K1. Recently, we reported that P2K1 interacts with Integrin-linked kinase 5 (ILK5), a Raf-like MAPKKK protein, and phosphorylates ILK5 to regulate purinergic signaling in relation to plant innate immunity. Here, we report that P2K2 also interacts with the ILK5 protein in planta. Furthermore, we demonstrate that P2K2 phosphorylates ILK5 in the presence of [γ-32P] ATP, similar to P2K1. However, unlike P2K1, P2K2 exhibits strong phosphorylation even when the Serine 192 residue of ILK5 is mutated to Alanine (ILK5^S192A), suggesting the possibility of phosphorylation of other residues to fully regulate ILK5 protein function.