Tyrosine phosphorylation has emerged as an important regulator of plasma membrane-localized immune receptors activity. Here， we investigate the role of tyrosine phosphorylation in the regulation of rice RESISTANCE 21 (XA21)-mediated immunity. We demonstrate that the juxtamembrane and kinase domain of -expressed XA21 (XA21JK) autophosphorylates on tyrosine residues. Directed mutagenesis of four out of the nine tyrosine residues in XA21JK reduced autophosphorylation. These sites include Tyr in the juxtamembrane domain， and Tyr， Tyr， and Tyr in the kinase domain. Rice plants expressing XA21-GFP fusion proteins or proteins with these tyrosine residues individually mutated to phenylalanine (XA21-GFP)， whi... More
Tyrosine phosphorylation has emerged as an important regulator of plasma membrane-localized immune receptors activity. Here， we investigate the role of tyrosine phosphorylation in the regulation of rice RESISTANCE 21 (XA21)-mediated immunity. We demonstrate that the juxtamembrane and kinase domain of -expressed XA21 (XA21JK) autophosphorylates on tyrosine residues. Directed mutagenesis of four out of the nine tyrosine residues in XA21JK reduced autophosphorylation. These sites include Tyr in the juxtamembrane domain， and Tyr， Tyr， and Tyr in the kinase domain. Rice plants expressing XA21-GFP fusion proteins or proteins with these tyrosine residues individually mutated to phenylalanine (XA21-GFP)， which prevents phosphorylation at these sites， maintain resistance to pv. . In contrast， plants expressing phosphomimetic XA21 variants with tyrosine mutated to aspartate (XA21-GFP) were susceptible. In vitro purified XA21JK， XA21JK， and XA21JK variants are catalytically active， whereas activity was not detected in XA21JK and the four XA21JK variants. We previously demonstrated that interaction of XA21 with the co-receptor OsSERK2 is critical for biological function. Four of the XA21JK variants maintain interaction with OsSERK2 as well as the XA21 binding (XB) proteins XB3 and XB15 in yeast， suggesting that these four tyrosine residues are not required for their interaction. Taken together， these results suggest that XA21 is capable of tyrosine autophosphorylation， but the identified tyrosine residues are not required for activation of XA21-mediated immunity or interaction with predicted XA21 signaling proteins.
