Identification of a C2H2-type zinc finger transcription factor (ZAT10) from Arabidopsis as a substrate of MAP kinase
Identification of a C2H2-type zinc finger transcription factor (ZAT10) from Arabidopsis as a substrate of MAP kinase
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DOI：
10.1007/s00299-011-1192-x
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发表时间：
2012-04
影响因子：
6.2
通讯作者：
X. Nguyen;S. H. Kim;Kyun Oh Lee;Kyung Eun Kim;Xiao-Min Liu;H. Han;M. Hoang;Shin-Woo Lee;
中科院分区：
文献类型：
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作者：
X. Nguyen;S. H. Kim;Kyun Oh Lee;Kyung Eun Kim;Xiao-Min Liu;H. Han;M. Hoang;Shin-Woo Lee;
丝裂原活化蛋白激酶（MAPKs或MPKs）是植物中最重要和最保守的信号分子之一。mpk可以通过磷酸化转录因子直接调控基因表达。然而，只有少数mpk的靶底物被分离出来。在这里，我们从拟南芥中鉴定出c2h2型锌指转录因子ZAT10作为MPKs的底物。通过体外和体内蛋白相互作用分析，我们证明ZAT10直接与MPK3和MPK6相互作用。在激酶实验中，ZAT10被重组拟南芥mpk3和MPK6磷酸化。此外，在凝胶激酶实验中，ZAT10也被从拟南芥植物中制备的天然MPK3和MPK6磷酸化。磷酸肽质谱分析用于确定ZAT10中两个MPK磷酸化位点。这些位点通过位点定向诱变和体外激酶试验得到验证。
Mitogen-activated protein kinases (MAPKs or MPKs) are one of the most important and conserved signaling molecules in plants. MPKs can directly modulate gene expression by the phosphorylation of transcription factors. However, only a few target substrates of MPKs have been isolated. Here, we identified a C2H2-type zinc finger transcription factor fromArabidopsis, ZAT10, as a substrate of MPKs. Using in vitro and in vivo protein–protein interaction analyses, we demonstrated that ZAT10 directly interacted with MPK3 and MPK6. ZAT10 was phosphorylated by recombinantArabidopsisMPK3 and MPK6 in a kinase assay. Furthermore, ZAT10 was also phosphorylated by native MPK3 and MPK6 prepared fromArabidopsisplants in an in-gel kinase assay. Mass spectrometry analysis of phosphopeptides was used to determine two MPK phosphorylation sites in ZAT10. These sites were verified by site-directed mutagenesis and in vitro kinase assays.