Leucine-rich repeat receptor-like protein kinase AtORPK1 promotes oxidative stress resistance in an AtORPK1-AtKAPP mediated module in Arabidopsis

Signal perception and transduction by the cell surface receptors are essential for cell-cell communication and plant response to abiotic stress. In this work, a previously uncharacterized leucine-rich repeat receptor-like kinase (LRR-RLK), Oxidative-stress Related Protein Kinase 1 (AtORPK1), was isolated from Arabidopsis thaliana, and its biological function was investigated in protoplasts, BY-2 cells and transgenic Arabidopsis plants. AtORPK1 is ubiquitously expressed in various tissues and organs of Arabidopsis at different developmental stages. Loss-of-function of AtORPK1 reduced, whereas overexpression of AtORPK1 increased, the oxidative stress resistance and oxidative stress responsive gene expression in orpk1 mutant and AtORPK1 transgenic Arabidopsis. Sub-cellular localization analyses revealed that AtORPK1 is localized to plasma membrane and endosomes, and the specific localization was significantly affected by hydrogen peroxide (H2O2) treatment. Further GFP, CFP, YFP and RFP fusion protein co-localization and FRET analyses demonstrated that AtORPK1 interacted and co-localized with AtKAPP, a common downstream phosphatase, in the enlarged endosomes such as prevacuolar compartments. Our results indicate that AtORPK1 functions as a positive molecular link between the oxidative stress signaling and antioxidant stress in plants.