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NO-inducible nitrosothionein mediates NO removal in tandem with thioredoxin

Nature Chemical Biology volume 9, pages 657–663 (2013)Cite this article

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Abstract

Nitric oxide (NO) is a toxic reactive nitrogen species that induces microbial adaption mechanisms. Screening a genomic DNA library identified a new gene, ntpA, that conferred growth tolerance upon Aspergillus nidulans against exogenous NO. The gene encoded a cysteine-rich 23-amino-acid peptide that reacted with NO and S-nitrosoglutathione to generate an S-nitrosated peptide. Disrupting ntpA increased amounts of cellular S-nitrosothiol and NO susceptibility. Thioredoxin and its reductase denitrosated the S-nitrosated peptide, decreased cellular S-nitrosothiol and conferred tolerance against NO, indicating peptide-mediated catalytic NO removal. The peptide binds copper(I) in vitro but is dispensable for metal tolerance in vivo. NO but not metal ions induced production of the peptide and ntpA transcripts. We discovered that the thionein family of peptides has NO-related functions and propose that the new peptide be named NO-inducible nitrosothionein (iNT). The ubiquitous distribution of iNT-like polypeptides constitutes a potent NO-detoxifying mechanism that is conserved among various organisms.

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Figure 1: Identification of NO-tolerating ntpA.
Figure 2: S-nitrosothionein and iNT have similar properties.
Figure 3: Denitrosation of iNT-SNO by Trx system.
Figure 4: Defense against heavy metals does not require iNT or MTLs.
Figure 5: Biochemical mechanism of iNT-dependent NO detoxification.

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Acknowledgements

We thank H. Shoun and S. Fushinobu for help with fluorometry and S. Osmani for his valuable suggestions in fungal transformation. We thank N. Foster for helpful discussion and critical reading of the manuscript. This study was supported by Grants-in-Aid for Scientific Research, Japan Society for the Promotion of Science (23-01090 to S.Z. and 21380055 to N.T.).

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  1. Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan

    Shengmin Zhou, Toshiaki Narukami, Shunsuke Masuo, Motoyuki Shimizu, Tomoya Fujita, Yuki Doi, Yosuke Kamimura & Naoki Takaya

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Contributions

N.T. and S.Z. planned the studies and prepared the manuscript. S.Z., T.N. and Y.K. designed and performed experiments. S.M., T.N., M.S., T.F., Y.D. and Y.K. performed the experiments.

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Correspondence to Naoki Takaya.

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Zhou, S., Narukami, T., Masuo, S. et al. NO-inducible nitrosothionein mediates NO removal in tandem with thioredoxin. Nat Chem Biol 9, 657–663 (2013). https://doi.org/10.1038/nchembio.1316

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