StCDPK3 Phosphorylates In Vitro Two Transcription Factors Involved in GA and ABA Signaling in Potato: StRSG1 and StABF1.
StCDPK3 Phosphorylates In Vitro Two Transcription Factors Involved in GA and ABA Signaling in Potato: StRSG1 and StABF1.
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DOI：
10.1371/journal.pone.0167389
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Ulloa RM
中科院分区：
文献类型：
--
作者：
Grandellis C;Fantino E;Muñiz García MN;Bialer MG;Santin F;Capiati DA;Ulloa RM
钙依赖蛋白激酶，CDPKs，在植物中解码钙(Ca~(2+))瞬变并启动下游反应。为了了解CDPKs是如何影响植物生理的，必须确定其特定的靶蛋白。在烟草中，调控赤霉素(GA)含量的bZIP转录因子抑制芽生长(NtRSG)是NtCDPK1的一个特异靶点。马铃薯StCDPK3与NtCDPK1在N端可变区也有88%的同源性。在这项工作中，我们观察到NtRSG也被StCDPK3磷酸化。马铃薯RSG转录因子家族由三个具有相似功能的成员组成。扩增并测序了与NtRSG最接近的同源物，命名为StRSG1。QRT-PCR结果表明，StRSG1主要在叶柄、茎、侧芽和根中表达。此外，GA处理还影响了StRSG1的表达。在叶片、叶柄、匍匐茎、根和休眠块茎中检测到StCDPK3的转录，并且转录水平对GA的响应发生了改变。构建了重组StRSG1-GST蛋白，并以此为底物进行了检测。6xHisStCDPK3能够以钙离子依赖的方式磷酸化马铃薯StRSG1，而6xHisStCDPK1不能。EMSA和磷酸化实验表明，StCDPK3还与参与ABA信号转导的转录因子StABF1(Abre结合因子1)相互作用并使其磷酸化。StABF1转录本主要在根、茎和匍匐茎中检测到。我们的数据表明，StCDPK3可能参与了块茎发育开始时ABA和GA信号之间的串扰。
Calcium-dependent protein kinases, CDPKs, decode calcium (Ca2+) transients and initiate downstream responses in plants. In order to understand how CDPKs affect plant physiology, their specific target proteins must be identified. In tobacco, the bZIP transcription factor Repression of Shoot Growth (NtRSG) that modulates gibberellin (GA) content is a specific target of NtCDPK1. StCDPK3 from potato is homologous (88% identical) to NtCDPK1 even in its N-terminal variable domain. In this work, we observe that NtRSG is also phosphorylated by StCDPK3. The potato RSG family of transcription factors is composed of three members that share similar features. The closest homologue to NtRSG, which was named StRSG1, was amplified and sequenced. qRT-PCR data indicate that StRSG1 is mainly expressed in petioles, stems, lateral buds, and roots. In addition, GA treatment affected StRSG1 expression. StCDPK3 transcripts were detected in leaves, petioles, stolons, roots, and dormant tubers, and transcript levels were modified in response to GA. The recombinant StRSG1-GST protein was produced and tested as a substrate for StCDPK3 and StCDPK1. 6xHisStCDPK3 was able to phosphorylate the potato StRSG1 in a Ca2+-dependent way, while 6xHisStCDPK1 could not. StCDPK3 also interacts and phosphorylates the transcription factor StABF1 (ABRE binding factor 1) involved in ABA signaling, as shown by EMSA and phosphorylation assays. StABF1 transcripts were mainly detected in roots, stems, and stolons. Our data suggest that StCDPK3 could be involved in the cross-talk between ABA and GA signaling at the onset of tuber development.
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