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Structural basis for redox regulation of Yap1 transcription factor localization

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

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Abstract

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.

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Acknowledgements

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

Affiliations

  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

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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.

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DOI

https://doi.org/10.1038/nature02790

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