Identification of a C2H2-type zinc finger transcription factor (ZAT10) from Arabidopsis as a substrate of MAP kinase

Abstract

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 C(2)H(2)-type zinc finger transcription factor from Arabidopsis, 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 recombinant Arabidopsis MPK3 and MPK6 in a kinase assay. Furthermore, ZAT10 was also phosphorylated by native MPK3 and MPK6 prepared from Arabidopsis plants 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.

The Heat Shock Factor A4A Confers Salt Tolerance and Is Regulated by Oxidative Stress and the Mitogen-Activated Protein Kinases MPK3 and MPK6

“…MAP kinases were reported to phosphorylate other transcription factors such as WRKY33, which is involved in regulation of ethylene biosynthesis (Li et al, 2012), or MYB44, which regulates abscisic acid sensitivity (Nguyen et al, 2012b). MPK3 and MPK6 can form in vivo complexes with ZAT10/salt tolerance zinc finger, regulating plant defense responses Nguyen et al, 2012a), while under salt and osmotic stress, MPK6 interacts with and phosphorylates ZAT6 (Liu et al, 2013).…”

A Mitogen-Activated Protein Kinase Cascade Module, MKK3-MPK6 and MYC2, Is Involved in Blue Light-Mediated Seedling Development in Arabidopsis

“…The bHLH transcription factor SPEECHLESS was found to be an in vitro substrate for phosphorylation by the MAPKs (MPK3 and MPK6) (Lampard et al, 2008). ZAT10, a C2H2-type zinc finger transcription factor from Arabidopsis, was identified as a substrate of Arabidopsis MPK3 and MPK6 (MPK3/6) (Nguyen et al, 2012). Recently, it has been reported in rice that a salt-responsive transcription factor, SERF1, binds to the promoter of MAP3K6 and MAPK5 and that SERF1 is also a phosphorylation target of MAPK5 (Schmidt et al, 2013).…”

The Cysteine2/Histidine2-Type Transcription Factor ZINC FINGER OF ARABIDOPSIS THALIANA6 Modulates Biotic and Abiotic Stress Responses by Activating Salicylic Acid-Related Genes and C-REPEAT-BINDING FACTOR Genes in Arabidopsis

“…ZFPs have been classified into at least nine types based on the number and location of characteristic residues (Cys and His), including C 2 H 2 , C 2 HC, C 2 HC 5 , C 3 H, C 3 HC 4 , C 4 , C 4 HC 3 , C 6 , and C 8 (Nguyen et al, 2012;Bogamuwa and Jang, 2013;Zhou et al, 2013). In Arabidopsis (Arabidopsis thaliana), the involvement of C 2 H 2 -type ZFPs, including Arabidopsis Zinc-Finger Protein1 (AZF1), AZF2, AZF3, Drought induced19 (Di19), Zinc Finger of Arabidopsis6 (ZAT6), ZAT7, ZAT10, and ZAT12, in plant abiotic stress response has been revealed (Liu et al, 2013b(Liu et al, , 2013d.…”

“…AtZAT10 played a dual role in response to abiotic stress because both gain-and loss-of-function mutations of AtZAT10 displayed enhanced resistance to drought and osmotic and salt stresses (Mittler et al, 2006). Recently, AtZAT10 was identified as a substrate of mitogenactivated protein kinase (Nguyen et al, 2012). CiftciYilmaz et al (2007) revealed that AtZAT7 positively mediated salt stress tolerance via modulating expression of several defense-responsive genes and that the ethylene-responsive element-binding factor-associated amphiphilic repression motif is required for abiotic stress response.…”