Arabidopsis MAP kinase phosphatase 1 (AtMKP1) is a member of the mitogen-activated protein kinase (MPK) phosphatase family, which negatively regulates AtMPKs. We have previously shown that AtMKP1 is regulated by calmodulin (CaM). Here, we examined the phosphorylation of AtMKP1 by its substrate AtMPK6. Intriguingly, AtMKP1 was phosphorylated by AtMPK6, one of AtMKP1 substrates. Four phosphorylation sites were identified by phosphoamino acid analysis, TiO2 chromatography and mass spectrometric analysis. Site-directed mutation of these residues in AtMKP1 abolished the phosphorylation by AtMPK6. In addition, AtMKP1 interacted with AtMPK6 as demonstrated by the yeast two-hybrid system. Finally, the phosphatase activity of AtMKP1 increased approximately twofold following phosphorylation by AtMPK6. By in-gel kinase assays, we showed that AtMKP1 could be rapidly phosphorylated by AtMPK6 in plants. Our results suggest that the catalytic activity of AtMKP1 in plants can be regulated not only by Ca2+/CaM, but also by its physiological substrate, AtMPK6.
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We thank Dr. Hans J. Bohnert for critical reading and insightful comments. This work was supported by grants from World Class University program (R32-10148) and Basic Science Research Program (2010-0010607) of National Research Foundation (NRF) funded by MOEST, Korea. KEK was supported by scholarships from the BK21 program of the Ministry of Education, Science and Technology, Korea.
H. C. Park, E. H. Song, and X. C. Nguyen contributed equally to this work.
Communicated by J. R. Liu.
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Park, H.C., Song, E.H., Nguyen, X.C. et al. Arabidopsis MAP kinase phosphatase 1 is phosphorylated and activated by its substrate AtMPK6. Plant Cell Rep 30, 1523–1531 (2011). https://doi.org/10.1007/s00299-011-1064-4
- Mitogen-activated protein kinase (MPK)
- Mitogen-activated protein kinase phosphatase (MKP)