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Ca2+/Calmodulin Activates an MAP Kinase Through the Inhibition of a Protein Phosphatase (DsPTP1) in Arabidopsis

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Abstract

Mitogen-activated protein kinases (MPKs) play roles as critical signal components in the environmental stress responses and developmental processes in plants. Calcium ion (Ca2+) is one of the most essential ubiquitous intracellular second messengers involved in many signal transduction pathways in plants. It was previously known that MPKs are activated by the increasing Ca2+ concentration. However, the mechanism of how Ca2+ activates MPKs is not elucidated yet. In this study, we revealed that Ca2+ could activate MPK signaling pathway via inhibiting the activity of a dual-specificity protein phosphatase1 (DsPTP1) by Ca2+/calmodulin (CaM). We showed that DsPTP1 directly interacts with MPK6 in vitro and in vivo. DsPTP1 was able to inactivate the active MPK6 by dephosphorylation. Interestingly, the DsPTP1-mediated dephosphorylation of MPK6 was strongly inhibited by Ca2+/CaM. Moreover, this inhibition was caused by the binding of CaM to the calmodulin-binding domain II (CaMBDII) of DsPTP1. This study implies that Ca2+/CaM is involved in the activation of MPKs through the inhibition of DsPTP1.

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Acknowledgements

This work was supported by a grant from the Cooperative Research Program for Agriculture Science and Technology Development (No. PJ01590901) funded by the Rural Development Administration, Republic of Korea and by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (Nos. 2021R1I1A1A01040693 and 2020R1A6A1A03044344), and partly supported by the National Institute of Ecology (NIE-C-2021-15).

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Author notes

  1. Kyung Eun Kim and Nhan Thi Nguyen have contributed equally to this work.

Authors and Affiliations

  1. Division of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, 52828, Republic of Korea

    Kyung Eun Kim, Nhan Thi Nguyen, Sun Ho Kim, Sunghwa Bahk, Mi Sun Cheong, Kyun Oh Lee, Jong Chan Hong & Woo Sik Chung

  2. Team of Vulnerable Ecological Research, Division of Climate and Ecology, Bureau of Conservation and Assessment Research, National Institute of Ecology, Seocheon, 33657, Republic of Korea

    Hyeong Cheol Park

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  1. Kyung Eun Kim
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  2. Nhan Thi Nguyen
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  3. Sun Ho Kim
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Contributions

KEK, NTN and WSC designed, planned and organized the experiments. KEK, NTN, SB and MSC performed biochemical experiments in this study. NTN, SHK, HCP, KOL, JCH and WSC analyzed data and wrote the manuscript with feedback from all authors.

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Correspondence to Woo Sik Chung.

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12374_2021_9338_MOESM1_ESM.pdf

Supplementary file1 Supplemental Fig. 1 Inactivation of MEK2-activated MPK6 by DsPTP1 was inhibited by Ca2+/CaM. A Inhibition of DsPTP1 by Ca2+/CaM. Activated MPK6 by MEK2 was incubated without or with 5 μg of DsPTP1, 0.5 mM Ca2+, 5 μg of CaM and 5 mM EGTA. The autophosphorylation activities (MPK6-P) and the substrate phosphorylation activities (MBP-P) of MPK6 were detected by in vitro kinase assays. B Inactivation of activated MPK6 by DsPTP1 is dependent on the concentration of Ca2+/CaM. Phosphorylated MPK6 by MEK2 (1 μg) were incubated with DsPTP1 (4 μg) in the presence of the different concentrations of CaM (1–5 μg). All reactions were performed in the presence of 0.5 mM Ca2+. The autophosphorylation activities (MPK6-P) and the substrate phosphorylation activities (MBP-P) of MPK6 were detected by in vitro kinase assays. (PDF 365 KB)

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Kim, K.E., Nguyen, N.T., Kim, S.H. et al. Ca2+/Calmodulin Activates an MAP Kinase Through the Inhibition of a Protein Phosphatase (DsPTP1) in Arabidopsis. J. Plant Biol. 65, 65–74 (2022). https://doi.org/10.1007/s12374-021-09338-x

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