Overexpression of Phosphomimic Mutated OsWRKY53 Leads to Enhanced Blast Resistance in Rice
Figure 2
Phosphorylation of OsWRKY53 does not alter its W-box binding ability.
A, W-box-specific DNA-binding activity of OsWRKY53. GMSA assay was performed using purified recombinant OsWRKY53 protein and 32P-labeled W-box probe containing the W-box cis-elements in the OsWRKY53 promoter. The specificity of the W-box binding activity was demonstrated by competition assay using 125-fold excess amount of unlabeled W-box probe (W) and mutated W-box probe (mW). B, Phosphorylation of OsWRKY53 does not enhance its W-box binding activity. Purified recombinant OsWRKY53 protein was phosphorylated using the OsMPK6 activated by OsMKK4DD. GMSA was performed as in A. C, W-box binding activity of OsWRKY53 variant proteins. Purified recombinant OsWRKY53, OsWRKY53SA and OsWRKY53SD proteins were subjected to GMSA. GMSA was performed as in A. WT, native His-OsWRKY53; SA, His-OsWRKY53SA; SD, His-OsWRKY53SD.