Stimulus-specific accumulation of second messengers like reactive oxygen species （ROS） and Ca^＋ are central to many signaling and regulation processes in plants. However, mechanisms that govern the reciprocal inter...Stimulus-specific accumulation of second messengers like reactive oxygen species （ROS） and Ca^＋ are central to many signaling and regulation processes in plants. However, mechanisms that govern the reciprocal interrelation of Ca^＋ and ROS signaling are only beginning to emerge. NADPH oxidases of the respiratory burst oxidase homolog （RBOH） family are critical components contributing to the generation of ROS while Calcineurin B-like （CBL） Ca^＋ sensor proteins together with their interacting kinases （CIPKs） have been shown to function in many Ca^＋- signaling processes. In this study, we identify direct functional interactions between both signaling systems. We report that the CBL-interacting pro- tein kinase ClPK26 specifically interacts with the N-terminal domain of RBOHF in yeast two-hybrid analyses and with the full-length RBOHF protein in plant cells. In addition, CIPK26 phosphorylates RBOHF in vitro and co-expression of either CBL1 or CBL9 with CIPK26 strongly enhances ROS production by RBOHF in HEK293T cells. Together, these findings identify a direct interconnection between CBL-ClPK-mediated Ca^＋ signaling and ROS signaling in plants and provide evidence for a synergistic activation of the NADPH oxidase RBOHF by direct Ca^＋-binding to its EF-hands and Ca2＋-induced phospho-rylation by CBL1/9-ClPK26 complexes.展开更多
文摘Stimulus-specific accumulation of second messengers like reactive oxygen species （ROS） and Ca^＋ are central to many signaling and regulation processes in plants. However, mechanisms that govern the reciprocal interrelation of Ca^＋ and ROS signaling are only beginning to emerge. NADPH oxidases of the respiratory burst oxidase homolog （RBOH） family are critical components contributing to the generation of ROS while Calcineurin B-like （CBL） Ca^＋ sensor proteins together with their interacting kinases （CIPKs） have been shown to function in many Ca^＋- signaling processes. In this study, we identify direct functional interactions between both signaling systems. We report that the CBL-interacting pro- tein kinase ClPK26 specifically interacts with the N-terminal domain of RBOHF in yeast two-hybrid analyses and with the full-length RBOHF protein in plant cells. In addition, CIPK26 phosphorylates RBOHF in vitro and co-expression of either CBL1 or CBL9 with CIPK26 strongly enhances ROS production by RBOHF in HEK293T cells. Together, these findings identify a direct interconnection between CBL-ClPK-mediated Ca^＋ signaling and ROS signaling in plants and provide evidence for a synergistic activation of the NADPH oxidase RBOHF by direct Ca^＋-binding to its EF-hands and Ca2＋-induced phospho-rylation by CBL1/9-ClPK26 complexes.
