Letter | Published:

Structural basis for redox regulation of Yap1 transcription factor localization

Nature volume 430, pages 917921 (19 August 2004) | Download Citation



The ability of organisms to alter their gene expression patterns in response to environmental changes is essential for viability. A central regulator of the response to oxidative stress in Saccharomyces cerevisiae is the Yap1 transcription factor. Upon activation by increased levels of reactive oxygen species, Yap1 rapidly redistributes to the nucleus where it regulates the expression of up to 70 genes1,2,3. Here we identify a redox-regulated domain of Yap1 and determine its high-resolution solution structure. In the active oxidized form, a nuclear export signal (NES) in the carboxy-terminal cysteine-rich domain is masked by disulphide-bond-mediated interactions with a conserved amino-terminal α-helix. Point mutations that weaken the hydrophobic interactions between the N-terminal α-helix and the C-terminal NES-containing domain abolished redox-regulated changes in subcellular localization of Yap1. Upon reduction of the disulphide bonds, Yap1 undergoes a change to an unstructured conformation that exposes the NES and allows redistribution to the cytoplasm. These results reveal the structural basis of redox-dependent Yap1 localization and provide a previously unknown mechanism of transcription factor regulation by reversible intramolecular disulphide bond formation.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1.

    , & Regulation of yAP-1 nuclear localization in response to oxidative stress. EMBO J. 16, 1710–1720 (1997)

  2. 2.

    et al. The H2O2 stimulon in Saccharomyces cerevisiae. J. Biol. Chem. 273, 22480–22489 (1998)

  3. 3.

    et al. Genomic expression programs in the response of yeast cells to environmental changes. Mol. Biol. Cell 11, 4241–4257 (2000)

  4. 4.

    , & Crm1p mediates regulated nuclear export of a yeast AP-1-like transcription factor. EMBO J. 17, 7416–7429 (1998)

  5. 5.

    , , & Nuclear import of the yeast AP-1-like transcription factor Yap1p is mediated by transport receptor Pse1p, and this import step is not affected by oxidative stress. J. Biol. Chem. 276, 21863–21869 (2001)

  6. 6.

    , , & Yap1p activates gene transcription in an oxidant-specific fashion. Mol. Cell. Biol. 19, 8302–8313 (1999)

  7. 7.

    , & The redox domain of the Yap1p transcription factor contains two disulfide bonds. Biochemistry 42, 11982–11991 (2003)

  8. 8.

    , & H2O2 sensing through oxidation of the Yap1 transcription factor. EMBO J. 19, 5157–5166 (2000)

  9. 9.

    , , , & A thiol peroxidase is an H2O2 receptor and redox-transducer in gene activation. Cell 111, 471–481 (2002)

  10. 10.

    et al. Mutual synergistic folding in recruitment of CBP/p300 by p160 nuclear receptor coactivators. Nature 415, 549–553 (2002)

  11. 11.

    et al. Structural basis of the redox switch in the OxyR transcription factor. Cell 105, 103–113 (2001)

  12. 12.

    et al. Activation of the redox-regulated molecular chaperone Hsp33–a two-step mechanism. Structure (Camb) 9, 377–387 (2001)

  13. 13.

    , , , & The 2.2 A crystal structure of Hsp33: a heat shock protein with redox-regulated chaperone activity. Structure (Camb) 9, 367–375 (2001)

  14. 14.

    & Regulation of nuclear localization: a key to a door. Annu. Rev. Cell Dev. Biol. 15, 291–339 (1999)

  15. 15.

    , & Nuclear transport and cancer: from mechanism to intervention. Nature Rev. Cancer 4, 106–117 (2004)

  16. 16.

    et al. Intramolecular masking of nuclear import signal on NF-AT4 by casein kinase I and MEKK1. Cell 93, 851–861 (1998)

  17. 17.

    et al. High-resolution structure of the oligomerization domain of p53 by multidimensional NMR. Science 265, 386–391 (1994)

  18. 18.

    et al. A leucine-rich nuclear export signal in the p53 tetramerization domain: regulation of subcellular localization and p53 activity by NES masking. EMBO J. 18, 1660–1672 (1999)

  19. 19.

    , , , & A zinc clasp structure tethers Lck to T cell coreceptors CD4 and CD8. Science 301, 1725–1728 (2003)

  20. 20.

    & A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122, 19–27 (1989)

  21. 21.

    , & Structure of Bax: coregulation of dimer formation and intracellular localization. Cell 103, 645–654 (2000)

  22. 22.

    et al. NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J. Biomol. NMR 6, 277–293 (1995)

  23. 23.

    , , & A common-sense approach to peak picking in 2-dimensional, 3-dimensional, and 4-dimensional spectra using automatic computer-analysis of contour diagrams. J. Magn. Reson. 95, 214–220 (1991)

  24. 24.

    & Multidimensional heteronuclear nuclear magnetic resonance of proteins. Methods Enzymol. 239, 349–363 (1994)

  25. 25.

    , , & Protein NMR Spectroscopy: Principles and Practice (Academic, San Diego, 1995)

  26. 26.

    , & Protein backbone angle restraints from searching a database for chemical shift and sequence homology. J. Biomol. NMR 13, 289–302 (1999)

  27. 27.

    , & Tunable alignment of macromolecules by filamentous phage yields dipolar coupling interactions. Nature Struct. Biol. 5, 1065–1074 (1998)

  28. 28.

    , , , & Use of dipolar 1H–15N and 1H–13C couplings in the structure determination of magnetically oriented macromolecules in solution. Nature Struct. Biol. 4, 732–738 (1997)

  29. 29.

    , , & The Xplor-NIH NMR molecular structure determination package. J. Magn. Reson. 160, 65–73 (2003)

Download references


We would like to thank E. Andrade for preparing the Yap1-RD expression construct, S. Moye-Rowley for providing reagents, C. Jackson for suggestions, C. Wu for use of the mass spectrometer and C. A. Combs for his expertise and advice regarding microscopy-related experiments. We also thank A. Gronenborn, R. Hegde, E. Komives, E. Korn, S. Moye-Rowley and W. Outten for critical reading of this manuscript. M.J.W. is supported by a Research Associateship from the National Research Council.

Author information


  1. Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892-5430, USA

    • Matthew J. Wood
    •  & Gisela Storz
  2. Laboratory of Biophysical Chemistry, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892-8013, USA

    • Nico Tjandra


  1. Search for Matthew J. Wood in:

  2. Search for Gisela Storz in:

  3. Search for Nico Tjandra in:

Competing interests

The authors declare that they have no competing financial interests.

Corresponding authors

Correspondence to Gisela Storz or Nico Tjandra.

Supplementary information

Word documents

  1. 1.

    Supplementary Figure 1

    In vivo analysis of full length wild-type GFP-Yap1 and Phe302A, Met306A and Val309A mutant derivatives.

  2. 2.

    Supplementary Table 1

    Yap1-RD NMR structure determination statistics.

About this article

Publication history






Further reading


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.