Ishida Sarahmi, Yuasa Takashi, Nakata Masaru, Takahashi Yohsuke
Department of Biological Sciences, Graduate School of Science, University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Plant Cell. 2008 Dec;20(12):3273-88. doi: 10.1105/tpc.107.057489. Epub 2008 Dec 23.
The homeostasis of gibberellins (GAs) is maintained by negative feedback in plants. REPRESSION OF SHOOT GROWTH (RSG) is a tobacco (Nicotiana tabacum) transcriptional activator that has been suggested to play a role in GA feedback by the regulation of GA biosynthetic enzymes. The 14-3-3 signaling proteins negatively regulate RSG by sequestering it in the cytoplasm in response to GAs. The phosphorylation on Ser-114 of RSG is essential for 14-3-3 binding of RSG. Here, we identified tobacco Ca(2+)-dependent protein kinase (CDPK1) as an RSG kinase that promotes 14-3-3 binding to RSG by phosphorylation of Ser-114 of RSG. CDPK1 interacts with RSG in a Ca(2+)-dependent manner in vivo and in vitro and specifically phosphorylates Ser-114 of RSG. Inhibition of CDPK repressed the GA-induced phosphorylation of Ser-114 of RSG and the GA-induced nuclear export of RSG. Overexpression of CDPK1 inhibited the feedback regulation of a GA 20-oxidase gene and resulted in sensitization to the GA biosynthetic inhibitor. Our results suggest that CDPK1 decodes the Ca(2+) signal produced by GAs and regulates the intracellular localization of RSG.
植物中赤霉素(GAs)的稳态通过负反馈得以维持。抑制茎生长(RSG)是一种烟草(Nicotiana tabacum)转录激活因子,有人认为它通过调节GA生物合成酶在GA反馈中发挥作用。14-3-3信号蛋白通过响应GAs将RSG隔离在细胞质中,从而对RSG进行负调控。RSG第114位丝氨酸的磷酸化对于RSG与14-3-3的结合至关重要。在此,我们鉴定出烟草钙依赖蛋白激酶(CDPK1)为一种RSG激酶,它通过对RSG第114位丝氨酸的磷酸化促进RSG与14-3-3的结合。CDPK1在体内和体外均以钙依赖的方式与RSG相互作用,并特异性地磷酸化RSG的第114位丝氨酸。抑制CDPK可抑制GA诱导的RSG第114位丝氨酸的磷酸化以及GA诱导的RSG核输出。过表达CDPK1会抑制GA 20-氧化酶基因的反馈调节,并导致对GA生物合成抑制剂敏感。我们的结果表明,CDPK1解码由GAs产生的钙信号,并调节RSG的细胞内定位。