Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Peroxiredoxin-2 and STAT3 form a redox relay for H2O2 signaling

Nature Chemical Biology volume 11, pages 64–70 (2015)Cite this article

Subjects

Abstract

Hydrogen peroxide (H2O2) acts as a signaling messenger by oxidatively modifying distinct cysteinyl thiols in distinct target proteins. However, it remains unclear how redox-regulated proteins, which often have low intrinsic reactivity towards H2O2 (kapp ∼1–10 M−1 s−1), can be specifically and efficiently oxidized by H2O2. Moreover, cellular thiol peroxidases, which are highly abundant and efficient H2O2 scavengers, should effectively eliminate virtually all of the H2O2 produced in the cell. Here, we show that the thiol peroxidase peroxiredoxin-2 (Prx2), one of the most H2O2-reactive proteins in the cell (kapp ∼107–108 M−1 s−1), acts as a H2O2 signal receptor and transmitter in transcription factor redox regulation. Prx2 forms a redox relay with the transcription factor STAT3 in which oxidative equivalents flow from Prx2 to STAT3. The redox relay generates disulfide-linked STAT3 oligomers with attenuated transcriptional activity. Cytokine-induced STAT3 signaling is accompanied by Prx2 and STAT3 oxidation and is modulated by Prx2 expression levels.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Prx2 forms disulfide exchange intermediates with STAT3.
Figure 2: Prx2 oxidizes STAT3.
Figure 3: Prx2 targets the DNA binding and trans-activating domains of STAT3.
Figure 4: Independent oxidation pathways lead to covalent dimers and tetramers.
Figure 5: IL6-type cytokines induce Prx2 and STAT3 oxidation.
Figure 6: Generalized model of peroxidase-mediated redox signaling.

Similar content being viewed by others

References

  1. D'Autréaux, B. & Toledano, M.B. ROS as signalling molecules: mechanisms that generate specificity in ROS homeostasis. Nat. Rev. Mol. Cell Biol. 8, 813–824 (2007).

    Article  Google Scholar 

  2. Finkel, T. Signal transduction by reactive oxygen species. J. Cell Biol. 194, 7–15 (2011).

    Article  CAS  Google Scholar 

  3. Winterbourn, C.C. Reconciling the chemistry and biology of reactive oxygen species. Nat. Chem. Biol. 4, 278–286 (2008).

    Article  CAS  Google Scholar 

  4. Winterbourn, C.C. & Hampton, M.B. Thiol chemistry and specificity in redox signaling. Free Radic. Biol. Med. 45, 549–561 (2008).

    Article  CAS  Google Scholar 

  5. Stone, J.R. & Yang, S. Hydrogen peroxide: a signaling messenger. Antioxid. Redox Signal. 8, 243–270 (2006).

    Article  CAS  Google Scholar 

  6. Woo, H.A. et al. Inactivation of peroxiredoxin I by phosphorylation allows localized H2O2 accumulation for cell signaling. Cell 140, 517–528 (2010).

    Article  CAS  Google Scholar 

  7. Delaunay, A., Pflieger, D., Barrault, M.B., Vinh, J. & Toledano, M.B. A thiol peroxidase is an H2O2 receptor and redox-transducer in gene activation. Cell 111, 471–481 (2002).

    Article  CAS  Google Scholar 

  8. Gutscher, M. et al. Proximity-based protein thiol oxidation by H2O2-scavenging peroxidases. J. Biol. Chem. 284, 31532–31540 (2009).

    Article  CAS  Google Scholar 

  9. Fomenko, D.E. et al. Thiol peroxidases mediate specific genome-wide regulation of gene expression in response to hydrogen peroxide. Proc. Natl. Acad. Sci. USA 108, 2729–2734 (2011).

    Article  CAS  Google Scholar 

  10. Zito, E. et al. Oxidative protein folding by an endoplasmic reticulum–localized peroxiredoxin. Mol. Cell 40, 787–797 (2010).

    Article  CAS  Google Scholar 

  11. Tavender, T.J., Springate, J.J. & Bulleid, N.J. Recycling of peroxiredoxin IV provides a novel pathway for disulphide formation in the endoplasmic reticulum. EMBO J. 29, 4185–4197 (2010).

    Article  CAS  Google Scholar 

  12. Wei, P.C. et al. Loss of the oxidative stress sensor NPGPx compromises GRP78 chaperone activity and induces systemic disease. Mol. Cell 48, 747–759 (2012).

    Article  CAS  Google Scholar 

  13. Jarvis, R.M., Hughes, S.M. & Ledgerwood, E.C. Peroxiredoxin 1 functions as a signal peroxidase to receive, transduce, and transmit peroxide signals in mammalian cells. Free Radic. Biol. Med. 53, 1522–1530 (2012).

    Article  CAS  Google Scholar 

  14. Sobotta, M.C. et al. Exposing cells to H2O2: a quantitative comparison between continuous low-dose and one-time high-dose treatments. Free Radic. Biol. Med. 60, 325–335 (2013).

    Article  CAS  Google Scholar 

  15. Lee, W. et al. Human peroxiredoxin 1 and 2 are not duplicate proteins: the unique presence of CYS83 in Prx1 underscores the structural and functional differences between Prx1 and Prx2. J. Biol. Chem. 282, 22011–22022 (2007).

    Article  CAS  Google Scholar 

  16. Li, L. & Shaw, P.E.A. STAT3 dimer formed by inter-chain disulphide bridging during oxidative stress. Biochem. Biophys. Res. Commun. 322, 1005–1011 (2004).

    Article  CAS  Google Scholar 

  17. Li, L., Cheung, S.H., Evans, E.L. & Shaw, P.E. Modulation of gene expression and tumor cell growth by redox modification of STAT3. Cancer Res. 70, 8222–8232 (2010).

    Article  CAS  Google Scholar 

  18. Peskin, A.V. et al. The high reactivity of peroxiredoxin 2 with H2O2 is not reflected in its reaction with other oxidants and thiol reagents. J. Biol. Chem. 282, 11885–11892 (2007).

    Article  CAS  Google Scholar 

  19. Ushio-Fukai, M. Localizing NADPH oxidase-derived ROS. Sci. STKE 2006, re8 (2006).

    PubMed  Google Scholar 

  20. Veal, E.A., Ross, S.J., Malakasi, P., Peacock, E. & Morgan, B.A. Ybp1 is required for the hydrogen peroxide–induced oxidation of the Yap1 transcription factor. J. Biol. Chem. 278, 30896–30904 (2003).

    Article  CAS  Google Scholar 

  21. Calvo, I.A. et al. Dissection of a redox relay: H2O2-dependent activation of the transcription factor Pap1 through the peroxidatic Tpx1-thioredoxin cycle. Cell Reports 5, 1413–1424 (2013).

    Article  CAS  Google Scholar 

  22. Choi, M.H. et al. Regulation of PDGF signalling and vascular remodelling by peroxiredoxin II. Nature 435, 347–353 (2005).

    Article  CAS  Google Scholar 

  23. Ng, D.C. et al. Stat3 regulates microtubules by antagonizing the depolymerization activity of stathmin. J. Cell Biol. 172, 245–257 (2006).

    Article  CAS  Google Scholar 

  24. Wegrzyn, J. et al. Function of mitochondrial Stat3 in cellular respiration. Science 323, 793–797 (2009).

    Article  CAS  Google Scholar 

  25. Tammineni, P. et al. The import of the transcription factor STAT3 into mitochondria depends on GRIM-19, a component of the electron transport chain. J. Biol. Chem. 288, 4723–4732 (2013).

    Article  CAS  Google Scholar 

  26. Okazaki, S., Naganuma, A. & Kuge, S. Peroxiredoxin-mediated redox regulation of the nuclear localization of Yap1, a transcription factor in budding yeast. Antioxid. Redox Signal. 7, 327–334 (2005).

    Article  CAS  Google Scholar 

  27. Nadeau, P.J., Charette, S.J., Toledano, M.B. & Landry, J. Disulfide bond-mediated multimerization of Ask1 and its reduction by thioredoxin-1 regulate H2O2-induced c-Jun NH2-terminal kinase activation and apoptosis. Mol. Biol. Cell 18, 3903–3913 (2007).

    Article  CAS  Google Scholar 

  28. Putker, M. et al. Redox-dependent control of FOXO/DAF-16 by transportin-1. Mol. Cell 49, 730–742 (2013).

    Article  CAS  Google Scholar 

  29. Myers, M.P. et al. TYK2 and JAK2 are substrates of protein-tyrosine phosphatase 1B. J. Biol. Chem. 276, 47771–47774 (2001).

    Article  CAS  Google Scholar 

  30. Mamoon, N.M. et al. Multiple cysteine residues are implicated in janus kinase 2-mediated catalysis. Biochemistry 46, 14810–14818 (2007).

    Article  CAS  Google Scholar 

  31. Godoy, J.R. et al. Redox atlas of the mouse. Immunohistochemical detection of glutaredoxin-, peroxiredoxin-, and thioredoxin-family proteins in various tissues of the laboratory mouse. Biochim. Biophys. Acta 1810, 2–92 (2011).

    Article  CAS  Google Scholar 

  32. Xu, G. et al. Thioredoxin-interacting protein regulates insulin transcription through microRNA-204. Nat. Med. 19, 1141–1146 (2013).

    Article  CAS  Google Scholar 

  33. Morgan, B., Sobotta, M.C. & Dick, T.P. Measuring EGSH and H2O2 with roGFP2-based redox probes. Free Radic. Biol. Med. 51, 1943–1951 (2011).

    Article  CAS  Google Scholar 

  34. Wang, X., Li, X. & Li, Y. A modified Coomassie Brilliant Blue staining method at nanogram sensitivity compatible with proteomic analysis. Biotechnol. Lett. 29, 1599–1603 (2007).

    Article  CAS  Google Scholar 

  35. Boersema, P.J., Raijmakers, R., Lemeer, S., Mohammed, S. & Heck, A.J. Multiplex peptide stable isotope dimethyl labeling for quantitative proteomics. Nat. Protoc. 4, 484–494 (2009).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

T.P.D. is supported by the Deutsche Forschungsgemeinschaft (SFB 1036, SFB 938 and SPP 1710). M.C.S. was supported by a PhD fellowship from the Boehringer Ingelheim Fonds. We thank B. Morgan and R. Jarvis for critical and helpful comments on the manuscript.

Author information

Authors and Affiliations

  1. Division of Redox Regulation, German Cancer Research Center (DKFZ), DKFZ-ZMBH Alliance, Heidelberg, Germany

    Mirko C Sobotta, Willy Liou, Sarah Stöcker, Deepti Talwar, Michael Oehler & Tobias P Dick

  2. Core facility for Mass Spectrometry and Proteomics, Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ-ZMBH Alliance, Heidelberg, Germany

    Thomas Ruppert & Annette N D Scharf

Authors
  1. Mirko C Sobotta
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Willy Liou
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sarah Stöcker
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Deepti Talwar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Michael Oehler
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Thomas Ruppert
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Annette N D Scharf
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Tobias P Dick
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

M.C.S. and T.P.D. conceived the project, designed the experiments, analyzed the data and wrote the manuscript. M.C.S. performed most experiments; W.L. generated and analyzed cysteine mutants; S.S. performed luciferase reporter assays; S.S. and D.T. analyzed cytokine-induced protein oxidation; M.O. performed experiments relating to the role of thioredoxin; T.R. and A.N.D.S. performed MS experiments.

Corresponding author

Correspondence to Tobias P Dick.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Results and Supplementary Figures 1–13. (PDF 2329 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sobotta, M., Liou, W., Stöcker, S. et al. Peroxiredoxin-2 and STAT3 form a redox relay for H2O2 signaling. Nat Chem Biol 11, 64–70 (2015). https://doi.org/10.1038/nchembio.1695

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nchembio.1695

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing