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 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.
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This work was supported by grants from the World Class University program (R32-10148) and the Basic Science Research Program (2010-0010607) of the National Research Foundation (NRF) funded by MOEST, and partly by a grant from the Next-Generation BioGreen 21 Program; (#PJ008173); and Agenda Project of RDA (No. PJ006681) funded by the Rural Development Administration, Republic of Korea. XCN, XML, KEK and HJH were supported by scholarships from the BK21 program funded by the MOEST in Korea.
Communicated by J. R. Liu.
X. C. Nguyen and S. H. Kim contributed equally to the article.
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Nguyen, X.C., Kim, S.H., Lee, K. et al. Identification of a C2H2-type zinc finger transcription factor (ZAT10) from Arabidopsis as a substrate of MAP kinase. Plant Cell Rep 31, 737–745 (2012). https://doi.org/10.1007/s00299-011-1192-x