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.

  • Letter
  • Published:

Redox-mediated substrate recognition by Sdp1 defines a new group of tyrosine phosphatases

Nature volume 447, pages 487–492 (2007)Cite this article

Abstract

Reactive oxygen species trigger cellular responses by activation of stress-responsive mitogen-activated protein kinase (MAPK) signalling pathways1,2. Reversal of MAPK activation requires the transcriptional induction of specialized cysteine-based phosphatases that mediate MAPK dephosphorylation3. Paradoxically, oxidative stresses generally inactivate cysteine-based phosphatases by thiol modification and thus could lead to sustained or uncontrolled MAPK activation4,5. Here we describe how the stress-inducible MAPK phosphatase, Sdp1, presents an unusual solution to this apparent paradox by acquiring enhanced catalytic activity under oxidative conditions. Structural and biochemical evidence reveals that Sdp1 employs an intramolecular disulphide bridge and an invariant histidine side chain to selectively recognize a tyrosine-phosphorylated MAPK substrate. Optimal activity critically requires the disulphide bridge, and thus, to the best of our knowledge, Sdp1 is the first example of a cysteine-dependent phosphatase that couples oxidative stress with substrate recognition. We show that Sdp1, and its paralogue Msg5, have similar properties and belong to a new group of phosphatases unique to yeast and fungal taxa.

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: Sdp1 activity is sensitive to reducing agents.
Figure 2: Sdp1 activity requires an intramolecular Cys 47–Cys 142 disulphide bridge.
Figure 3: Phosphotyrosine recognition by His 111 and the Cys 47–Cys 142 disulphide bridge.
Figure 4: Sdp1 and Msg5 are prototypic members of the WH phosphatase family.

Similar content being viewed by others

References

  1. Ikner, A. & Shiozaki, K. Yeast signaling pathways in the oxidative stress response. Mutat. Res. 569, 13–27 (2005)

    Article  CAS  Google Scholar 

  2. Chang, L. & Karin, M. Mammalian MAP kinase signalling cascades. Nature 410, 37–40 (2001)

    Article  ADS  CAS  Google Scholar 

  3. Keyse, S. M. Protein phosphatases and the regulation of mitogen-activated protein kinase signalling. Curr. Opin. Cell Biol. 12, 186–192 (2000)

    Article  CAS  Google Scholar 

  4. Tonks, N. K. Redox redux: revisiting PTPs and the control of cell signaling. Cell 121, 667–670 (2005)

    Article  CAS  Google Scholar 

  5. Salmeen, A. & Barford, D. Functions and mechanisms of redox regulation of cysteine-based phosphatases. Antioxid. Redox Signal. 7, 560–577 (2005)

    Article  CAS  Google Scholar 

  6. Barford, D., Das, A. K. & Egloff, M. P. The structure and mechanism of protein phosphatases: insights into catalysis and regulation. Annu. Rev. Biophys. Biomol. Struct. 27, 133–164 (1998)

    Article  CAS  Google Scholar 

  7. Alonso, A. et al. Protein tyrosine phosphatases in the human genome. Cell 117, 699–711 (2004)

    Article  CAS  Google Scholar 

  8. Pulido, R., Zuniga, A. & Ullrich, A. PTP-SL and STEP protein tyrosine phosphatases regulate the activation of the extracellular signal-regulated kinases ERK1 and ERK2 by association through a kinase interaction motif. EMBO J. 17, 7337–7350 (1998)

    Article  CAS  Google Scholar 

  9. Mattison, C. P., Spencer, S. S., Kresge, K. A., Lee, J. & Ota, I. M. Differential regulation of the cell wall integrity mitogen-activated protein kinase pathway in budding yeast by the protein tyrosine phosphatases Ptp2 and Ptp3. Mol. Cell. Biol. 19, 7651–7660 (1999)

    Article  CAS  Google Scholar 

  10. Rhee, S. G., Bae, Y. S., Lee, S. R. & Kwon, J. Hydrogen peroxide: a key messenger that modulates protein phosphorylation through cysteine oxidation. Sci. STKE 1 doi: 10.1126/stke.2000.53.pe1 (2000)

  11. Salmeen, A. et al. Redox regulation of protein tyrosine phosphatase 1B involves a sulphenyl-amide intermediate. Nature 423, 769–773 (2003)

    Article  ADS  CAS  Google Scholar 

  12. Seth, D. & Rudolph, J. Redox regulation of MAP kinase phosphatase 3. Biochemistry. 45,–8476–8487 (2006)

  13. Martin, H., Flandez, M., Nombela, C. & Molina, M. Protein phosphatases in MAPK signalling: we keep learning from yeast. Mol. Microbiol. 58, 6–16 (2005)

    Article  CAS  Google Scholar 

  14. Staleva, L., Hall, A. & Orlow, S. J. Oxidative stress activates FUS1 and RLM1 transcription in the yeast Saccharomyces cerevisiae in an oxidant-dependent manner. Mol. Biol. Cell 15, 5574–5582 (2004)

    Article  CAS  Google Scholar 

  15. Alic, N., Higgins, V. J., Pichova, A., Breitenbach, M. & Dawes, I. W. Lipid hydroperoxides activate the mitogen-activated protein kinase Mpk1p in Saccharomyces cerevisiae. J. Biol. Chem. 278, 41849–41855 (2003)

    Article  CAS  Google Scholar 

  16. Vilella, F., Herrero, E., Torres, J. & de la Torre-Ruiz, M. A. Pkc1 and the upstream elements of the cell integrity pathway in Saccharomyces cerevisiae, Rom2 and Mtl1, are required for cellular responses to oxidative stress. J. Biol. Chem. 280, 9149–9159 (2005)

    Article  CAS  Google Scholar 

  17. Krasley, E., Cooper, K. F., Mallory, M. J., Dunbrack, R. & Strich, R. Regulation of the oxidative stress response through Slt2p-dependent destruction of cyclin C in Saccharomyces cerevisiae. Genetics 172, 1477–1486 (2006)

    Article  CAS  Google Scholar 

  18. Levin, D. E. Cell wall integrity signaling in Saccharomyces cerevisiae. Microbiol. Mol. Biol. Rev. 69, 262–291 (2005)

    Article  CAS  Google Scholar 

  19. Hahn, J. S. & Thiele, D. J. Regulation of the Saccharomyces cerevisiae Slt2 kinase pathway by the stress-inducible Sdp1 dual specificity phosphatase. J. Biol. Chem. 277, 21278–21284 (2002)

    Article  CAS  Google Scholar 

  20. Collister, M. et al. YIL113w encodes a functional dual-specificity protein phosphatase which specifically interacts with and inactivates the Slt2/Mpk1p MAP kinase in S. cerevisiae. FEBS Lett. 527, 186–192 (2002)

    Article  CAS  Google Scholar 

  21. Groom, L. A., Sneddon, A. A., Alessi, D. R., Dowd, S. & Keyse, S. M. Differential regulation of the MAP, SAP and RK/p38 kinases by Pyst1, a novel cytosolic dual-specificity phosphatase. EMBO J. 15, 3621–3632 (1996)

    Article  CAS  Google Scholar 

  22. Todd, J. L., Tanner, K. G. & Denu, J. M. Extracellular regulated kinases (ERK) 1 and ERK2 are authentic substrates for the dual-specificity protein-tyrosine phosphatase VHR. A novel role in down-regulating the ERK pathway. J. Biol. Chem. 274, 13271–13280 (1999)

    Article  CAS  Google Scholar 

  23. Delauney, 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  Google Scholar 

  24. Jia, Z., Barford, D., Flint, A. J. & Tonks, N. K. Structural basis for phosphotyrosine peptide recognition by protein tyrosine phosphatase 1B. Science 268, 1754–1758 (1995)

    Article  ADS  CAS  Google Scholar 

  25. Stewart, A. E., Dowd, S., Keyse, S. M. & McDonald, N. Q. Crystal structure of the MAPK phosphatase Pyst1 catalytic domain and implications for regulated activation. Nature Struct. Biol. 6, 174–181 (1999)

    Article  CAS  Google Scholar 

  26. Kellis, M., Birren, B. W. & Lander, E. S. Proof and evolutionary analysis of ancient genome duplication in the yeast Saccharomyces cerevisiae. Nature 428, 617–624 (2004)

    Article  ADS  CAS  Google Scholar 

  27. Flandez, M., Cosano, I. C., Nombela, C., Martin, H. & Molina, M. Reciprocal regulation between Slt2 MAPK and isoforms of Msg5 dual-specificity protein phosphatase modulates the yeast cell integrity pathway. J. Biol. Chem. 279, 11027–11034 (2004)

    Article  CAS  Google Scholar 

  28. Kamata, H. et al. Reactive oxygen species promote TNFα-induced death and sustained JNK activation by inhibiting MAP kinase phosphatases. Cell 120, 649–661 (2005)

    Article  CAS  Google Scholar 

  29. Barford, D. The role of cysteine residues as redox-sensitive regulatory switches. Curr. Opin. Struct. Biol. 14, 679–686 (2004)

    Article  CAS  Google Scholar 

  30. Veal, E. A. et al. A 2-Cys peroxiredoxin regulates peroxide-induced oxidation and activation of a stress-activated MAP kinase. Mol. Cell 15, 129–139 (2004)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

G.C.F. was supported by a studentship from GlaxoSmithKline. N.Q.M. and S.M.K. are funded by CR-UK. M.J.R.S. is supported by BBSRC. We thank M. Toledano for experimental advice, T. Kuno for the Pmp1 plasmid and M. Way for critical comments on the manuscript. We also thank M. Cobb for the gift of the ERKpTpY expression vector, D. Thiele and J.-S. Hahn for yeast strains and Sdp1 expression plasmids, and A. Stewart, S. Tilley and J. Denu for assistance with early experiments.

Author Contributions G.C.F. and N.Q.M. determined the apo and sulphate-bound structures. M.S. determined the phosphotyrosine structure and carried out pNPP and ERK2 assays and blots. D.C.B. and N.Q.M. refined the structures. P.P.K. measured the free -SH content and reproduced all kinetic experiments. S.M.K. and M.J.R.S. designed the in vivo assays. M.C, M.J.D, V.M., and R.J.D. carried out in vivo assays using Myc-tagged Sdp1 proteins. S.H. and N.T carried out the mass spectrometry. S.M.K. and N.Q.M planned the project and designed the experiments. N.Q.M., S.M.K. and D.C.B. wrote the paper.

Coordinates have been deposited at the PDB with accession codes 2j16 (apo/sulphate-bound Sdp1) and 2j17 (phosphotyrosine–Sdp1 complex)

Author information

Author notes

  1. G. C. Fox

    Present address: Present address: ESRF, 6 Rue Jules Horowitz, 38043 Grenoble, Cedex 9, France.,

  2. G. C. Fox and M. Shafiq: These authors contributed equally to this work.

Authors and Affiliations

  1. Structural Biology Laboratory and,,

    G. C. Fox, M. Shafiq, D. C. Briggs, P. P. Knowles & N. Q. McDonald

  2. Protein Analysis Laboratory, The London Research Institute, Cancer Research UK, 44 Lincoln’s Inn Fields, London WC2A 3PX, UK,

    S. Hanrahan & N. Totty

  3. School of Crystallography, Birkbeck College, Malet Street, London WC1E 7HX, UK,

    N. Q. McDonald

  4. Cancer Research UK, Stress Response Laboratory, Biomedical Research Centre, Ninewells Hospital, Dundee DD1 9SY, UK,

    M. Collister, M. J. Didmon, R. J. Dickinson & S. M. Keyse

  5. Division of Gene Regulation and Expression, College of Life Sciences, MSI/WTB Complex, University of Dundee, Dundee DD1 5EH, UK,

    V. Makrantoni & M. J. R. Stark

Authors
  1. G. C. Fox
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Shafiq
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. D. C. Briggs
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. P. P. Knowles
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. Collister
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M. J. Didmon
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. V. Makrantoni
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. R. J. Dickinson
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. S. Hanrahan
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. N. Totty
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. M. J. R. Stark
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. S. M. Keyse
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. N. Q. McDonald
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to N. Q. McDonald.

Ethics declarations

Competing interests

Reprints and permissions information is available at www.nature.com/reprints. . The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Methods, Supplementary Discussion, Supplementary Figures S1-S3 with Legends and additional references. (PDF 1628 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fox, G., Shafiq, M., Briggs, D. et al. Redox-mediated substrate recognition by Sdp1 defines a new group of tyrosine phosphatases. Nature 447, 487–492 (2007). https://doi.org/10.1038/nature05804

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature05804

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

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