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
Global warming poses a serious threat to crops.Calcium-dependent protein kinases(CDPKs)/CPKs play vital roles in plant stress responses,but their exact roles in plant thermotolerance remains elusive.Here,we explored t...Global warming poses a serious threat to crops.Calcium-dependent protein kinases(CDPKs)/CPKs play vital roles in plant stress responses,but their exact roles in plant thermotolerance remains elusive.Here,we explored the roles of heat-induced ZmCDPK7 in thermotolerance in maize.ZmCDPK7-overexpressing maize plants displayed higher thermotolerance,photosynthetic rates,and antioxidant enzyme activity but lower H2 O2 and malondialdehyde(MDA)contents than wild-type plants under heat stress.ZmCDPK7-knockdown plants displayed the opposite patterns.ZmCDPK7 is attached to the plasma membrane but can translocate to the cytosol under heat stress.ZmCDPK7 interacts with the small heat shock protein sHSP17.4,phosphorylates sHSP17.4 at Ser-44 and the respiratory burst oxidase homolog RBOHB at Ser-99,and up regulates their expression.Site-directed mutagenesis of sHSP17.4 to generate a Ser-44-Ala substitution reduced ZmCDPK7’s enhancement of catalase activity but enhanced ZmCDPK7’s suppression of MDA accumulation in heat-stressed maize protoplasts.sHSP17.4,ZmCDPK7,and RBOHB were less strongly upregulated in response to heat stress in the abscisic acid-deficient mutant vp5 versus the wild type.Pretreatment with an RBOH inhibitor suppressed sHSP17.4 and ZmCDPK7 expression.Therefore,abscisic acid-induced ZmCDPK7 functions both upstream and downstream of RBOH and participates in thermotolerance in maize by mediating the phosphorylation of sHSP17.4,which might be essential for its chaperone function.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.
基金supported by the National Natural Science Foundation of Henan Joint Fund(U1604101 to XLH)the Key R&D and Promotion Projects of Henan Province(192102110009 to XLH,192102110004 to HY)。
文摘Global warming poses a serious threat to crops.Calcium-dependent protein kinases(CDPKs)/CPKs play vital roles in plant stress responses,but their exact roles in plant thermotolerance remains elusive.Here,we explored the roles of heat-induced ZmCDPK7 in thermotolerance in maize.ZmCDPK7-overexpressing maize plants displayed higher thermotolerance,photosynthetic rates,and antioxidant enzyme activity but lower H2 O2 and malondialdehyde(MDA)contents than wild-type plants under heat stress.ZmCDPK7-knockdown plants displayed the opposite patterns.ZmCDPK7 is attached to the plasma membrane but can translocate to the cytosol under heat stress.ZmCDPK7 interacts with the small heat shock protein sHSP17.4,phosphorylates sHSP17.4 at Ser-44 and the respiratory burst oxidase homolog RBOHB at Ser-99,and up regulates their expression.Site-directed mutagenesis of sHSP17.4 to generate a Ser-44-Ala substitution reduced ZmCDPK7’s enhancement of catalase activity but enhanced ZmCDPK7’s suppression of MDA accumulation in heat-stressed maize protoplasts.sHSP17.4,ZmCDPK7,and RBOHB were less strongly upregulated in response to heat stress in the abscisic acid-deficient mutant vp5 versus the wild type.Pretreatment with an RBOH inhibitor suppressed sHSP17.4 and ZmCDPK7 expression.Therefore,abscisic acid-induced ZmCDPK7 functions both upstream and downstream of RBOH and participates in thermotolerance in maize by mediating the phosphorylation of sHSP17.4,which might be essential for its chaperone function.
