Reactive sulfur species (RSS) modulate protein functions via S-polysulfidation of reactive Cys residues. Here, we report that Ca2+/calmodulin (CaM)-dependent protein kinase IV (CaMKIV) was reversibly inactivated by RSS via polysulfidation of the active-site Cys residue. CaMKIV is phosphorylated at Thr196 by its upstream CaMK kinase (CaMKK), resulting in the induction of its full activity. In vitro incubation of CaMKIV with the exogenous RSS donors Na2Sn (n = 2–4) resulted in dose-dependent inhibition of the CaMKK-induced phospho-Thr196 and consequent inactivation of the enzyme activity. Conversely, mutated CaMKIV (C198V) was refractory to the Na2Sn-induced enzyme inhibition. A biotin-polyethylene glycol-conjugated maleimide capture assay revealed that Cys198 in CaMKIV represents a target for S-polysulfidation. Furthermore, phosho-Thr196 and CaMKIV activity were inhibited by incubation with cysteine hydropersulfide, a newly identified RSS that is generated from cystine by cystathionine-γ-lyase. In transfected cells expressing CaMKIV, ionomycin-induced CaMKIV phosphorylation at Thr196 was decreased upon treatment with either Na2S4 or the endoplasmic reticulum (ER) stress inducer thapsigargin, whereas cells expressing mutant CaMKIV (C198V) were resistant to this treatment. In addition, the ionomycin-induced phospho-Thr196 of endogenous CaMKIV was also inhibited by treatment either with Na2S4 or thapsigargin in Jurkat T lymphocytes. Taken together, these data define a novel signaling function for intracellular RSS in inhibiting CaMKIV activity via S-polysulfidation of its Cys198 during the response to ER stress.
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August 2017
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In this issue of the Biochemical Journal, Zhu et al. (pages 2585–2599) report on the redox regulation of an SNF1-related protein kinase from Brassica napus. Their data suggest that it has potential role in signal transduction in B. napus guard cells.
Research Article|
July 18 2017
Reactive sulfur species inactivate Ca2+/calmodulin-dependent protein kinase IV via S-polysulfidation of its active-site cysteine residue
Tsuyoshi Takata;
Tsuyoshi Takata
1Department of Pharmacology, High Technology Research Center, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan
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Hideshi Ihara;
Hideshi Ihara
2Department of Biological Science, Graduate School of Science, Osaka Prefecture University, Sakai, Osaka 599-8531, Japan
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Naoya Hatano;
Naoya Hatano
3The Integrated Center for Mass Spectrometry, Graduate School of Medicine, Kobe University, Kobe 650-0017, Japan
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Yukihiro Tsuchiya;
Yukihiro Tsuchiya
1Department of Pharmacology, High Technology Research Center, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan
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Takaaki Akaike;
Takaaki Akaike
4Department of Environmental Health Sciences and Molecular Toxicology, Graduate School of Medicine, Tohoku University, Miyagi 980-8575, Japan
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Yasuo Watanabe
1Department of Pharmacology, High Technology Research Center, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan
Correspondence: Yasuo Watanabe (yasuwata@ac.shoyaku.ac.jp)
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Publisher: Portland Press Ltd
Received:
January 28 2017
Revision Received:
June 20 2017
Accepted:
June 21 2017
Accepted Manuscript online:
June 21 2017
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society
2017
Biochem J (2017) 474 (15): 2547–2562.
Article history
Received:
January 28 2017
Revision Received:
June 20 2017
Accepted:
June 21 2017
Accepted Manuscript online:
June 21 2017
Citation
Tsuyoshi Takata, Hideshi Ihara, Naoya Hatano, Yukihiro Tsuchiya, Takaaki Akaike, Yasuo Watanabe; Reactive sulfur species inactivate Ca2+/calmodulin-dependent protein kinase IV via S-polysulfidation of its active-site cysteine residue. Biochem J 1 August 2017; 474 (15): 2547–2562. doi: https://doi.org/10.1042/BCJ20170092
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