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Arabidopsis MAP kinase phosphatase 1 is phosphorylated and activated by its substrate AtMPK6

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Abstract

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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Acknowledgments

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.

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Authors and Affiliations

  1. Division of Applied Life Science (BK21 Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, 660-701, Republic of Korea

    Hyeong Cheol Park, Eun Hyeon Song, Xuan Canh Nguyen, Kyunghee Lee, Kyung Eun Kim, Ho Soo Kim, Sang Min Lee, Sun Ho Kim, Dae-Jin Yun & Woo Sik Chung

  2. Central Instrument Facility, Gyeongsang National University, Jinju, 660-701, Republic of Korea

    Dong Won Bae

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  1. Hyeong Cheol Park
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  2. Eun Hyeon Song
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  4. Kyunghee Lee
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Correspondence to Woo Sik Chung.

Additional information

Communicated by J. R. Liu.

H. C. Park, E. H. Song, and X. C. Nguyen contributed equally to this work.

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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

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  • DOI: https://doi.org/10.1007/s00299-011-1064-4

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