Hyperphosphorylation of a Mitochondrial Protein, Prohibitin, Is Induced by Calyculin A in a Rice Lesion-Mimic Mutant cdr1

Abstract The rice (Oryza sativa) lesion-mimic mutants, cell death and resistance (cdr), show spontaneous cell death on the entire leaf and exhibited significant resistance to the rice blast fungus. Our previous studies showed that CDR1 and CDR2 genes negatively regulated the phosphorylation steps leading to the activation of NADPH oxidase, which is associated with oxidative burst. To identify novel factors involved in the phosphorylation steps, the phosphorylation level of total proteins was compared between cdr mutants and wild type using two-dimensional gel electrophoresis. Here, we show that the phosphorylation level of four proteins in cdr1 was increased as compared with the wild type after calyculin A treatment. Partial amino acid sequences revealed that one of the four proteins is homologous to prohibitin (PHB), which has been shown to be associated with senescence and cell death and to function as a chaperone in the assembly of mitochondrial respiratory chain complex in yeast and mammals. Analysis of green fluorescent protein fusions indicated that rice PHB (OsPHB1) was targeted to mitochondria as found in yeast and mammals, suggesting a possibility that PHB is involved in defense response and/or programmed cell death through the mitochondrial function.