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The PerR transcription factor senses H2O2 by metal-catalysed histidine oxidation

Abstract

The sensing of reactive oxygen species is essential for cellular responses to oxidative stress1. The sensing of peroxides is typically mediated by redox-active cysteines in sensors such as the bacterial OxyR, OhrR, and Hsp33 proteins2,3. Bacillus subtilis PerR is the prototype for a widespread family of metal-dependent peroxide sensors that regulate inducible peroxide-defence genes4. Here we show that PerR senses peroxides by metal-catalysed oxidation. PerR contains two metal-binding sites: a structural Zn2+ site and a regulatory divalent metal ion site that preferentially binds Fe2+ or Mn2+ (ref. 5). Protein oxidation, catalysed by a bound ferrous ion, leads to the rapid and direct incorporation of one oxygen atom into histidine 37 (H37) or H91, two of the residues that coordinate the bound Fe2+. This mechanism accounts for the ability of PerR to sense low levels of hydrogen peroxide in vivo. The reduction of hydrogen peroxide by metal ions to generate highly reactive hydroxyl radicals underlies the genotoxic effects of peroxides1, and has been shown to contribute to enzyme inactivation, but has not previously been shown to provide a regulatory mechanism for peroxide sensing.

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Figure 1: PerR contains two metal-binding sites.
Figure 2: Metal-catalysed oxidation of PerR.
Figure 3: Oxidative inactivation of PerR:Zn,Fe and PerR:Zn,Mn.
Figure 4: Oxidation of PerR is localized to H37 and H91.

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Acknowledgements

We thank M. Fuangthong and S. Soonsanga for their initial analyses of functionally altered perR variants, E. Madsen for help with the anaerobic experiments, S. Zhang and R. Sherwood for assistance with mass spectrometry, and G. Storz, P. Kiley, J. Imlay, and C. M. Moore for comments. This work was supported by grants from the NSF and NIH. Author Contributions: J.-W.L. performed all experimental work, J.-W.L. and J.D.H. analysed results and co-wrote the paper.

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Lee, JW., Helmann, J. The PerR transcription factor senses H2O2 by metal-catalysed histidine oxidation. Nature 440, 363–367 (2006). https://doi.org/10.1038/nature04537

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