Skip to main content

Thank you for visiting 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.

Tyrosine dephosphorylation of H2AX modulates apoptosis and survival decisions


Life and death fate decisions allow cells to avoid massive apoptotic death in response to genotoxic stress. Although the regulatory mechanisms and signalling pathways controlling DNA repair and apoptosis are well characterized, the precise molecular strategies that determine the ultimate choice of DNA repair and survival or apoptotic cell death remain incompletely understood. Here we report that a protein tyrosine phosphatase, EYA, is involved in promoting efficient DNA repair rather than apoptosis in response to genotoxic stress in mammalian embryonic kidney cells by executing a damage-signal-dependent dephosphorylation of an H2AX carboxy-terminal tyrosine phosphate (Y142). This post-translational modification determines the relative recruitment of either DNA repair or pro-apoptotic factors to the tail of serine phosphorylated histone H2AX (γ-H2AX) and allows it to function as an active determinant of repair/survival versus apoptotic responses to DNA damage, revealing an additional phosphorylation-dependent mechanism that modulates survival/apoptotic decisions during mammalian organogenesis.

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

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: Loss of EYA leads to increased γ-H2AX-posititve apoptotic cells.
Figure 2: EYA interacts with H2AX in a DNA-damage-dependent manner.
Figure 3: EYA3 phosphorylation by ATM/ATR DNA-damage-dependent kinases regulates the interaction between EYA and H2AX.
Figure 4: Tyrosine phosphorylated H2AX is a substrate for EYA phosphatase.
Figure 5: H2AX Y142 phosphorylation discriminates between apoptotic and repair responses to DNA damage.


  1. Kumar, J. P. Signalling pathways in Drosophila and vertebrate retinal development. Nature Rev. Genet. 2, 846–857 (2001)

    CAS  Article  Google Scholar 

  2. Silver, S. J. & Rebay, I. Signaling circuitries in development: insights from the retinal determination gene network. Development 132, 3–13 (2005)

    CAS  Article  Google Scholar 

  3. Pignoni, F. et al. The eye-specification proteins So and Eya form a complex and regulate multiple steps in Drosophila eye development. Cell 91, 881–891 (1997)

    CAS  Article  Google Scholar 

  4. Bonini, N. M., Leiserson, W. M. & Benzer, S. The eyes absent gene: genetic control of cell survival and differentiation in the developing Drosophila eye. Cell 72, 379–395 (1993)

    CAS  Article  Google Scholar 

  5. Xu, P. X. et al. Eya1-deficient mice lack ears and kidneys and show abnormal apoptosis of organ primordia. Nature Genet. 23, 113–117 (1999)

    CAS  Article  Google Scholar 

  6. Li, X. et al. Eya protein phosphatase activity regulates Six1–Dach–Eya transcriptional effects in mammalian organogenesis. Nature 426, 247–254 (2003)

    ADS  CAS  Article  Google Scholar 

  7. Rayapureddi, J. P. et al. Eyes absent represents a class of protein tyrosine phosphatases. Nature 426, 295–298 (2003)

    ADS  CAS  Article  Google Scholar 

  8. Tootle, T. L. et al. The transcription factor Eyes absent is a protein tyrosine phosphatase. Nature 426, 299–302 (2003)

    ADS  CAS  Article  Google Scholar 

  9. Rayapureddi, J. P. et al. Characterization of a plant, tyrosine-specific phosphatase of the aspartyl class. Biochemistry 44, 751–758 (2005)

    CAS  Article  Google Scholar 

  10. Lu, C. et al. Cell apoptosis: requirement of H2AX in DNA ladder formation, but not for the activation of caspase-3. Mol. Cell 23, 121–132 (2006)

    CAS  Article  Google Scholar 

  11. Bassing, C. H. & Alt, F. W. The cellular response to general and programmed DNA double strand breaks. DNA Repair 3, 781–796 (2004)

    CAS  Article  Google Scholar 

  12. Bassing, C. H. et al. Increased ionizing radiation sensitivity and genomic instability in the absence of histone H2AX. Proc. Natl Acad. Sci. USA 99, 8173–8178 (2002)

    ADS  CAS  Article  Google Scholar 

  13. Karagiannis, T. C. & El-Osta, A. Chromatin modifications and DNA double-strand breaks: the current state of play. Leukemia 21, 195–200 (2007)

    CAS  Article  Google Scholar 

  14. van Attikum, H. & Gasser, S. M. The histone code at DNA breaks: a guide to repair? Nature Rev. Mol. Cell Biol. 6, 757–765 (2005)

    CAS  Article  Google Scholar 

  15. Lee, Y. M. et al. Determination of hypoxic region by hypoxia marker in developing mouse embryos in vivo: a possible signal for vessel development. Dev. Dyn. 220, 175–186 (2001)

    CAS  Article  Google Scholar 

  16. Haase, V. H. Hypoxia-inducible factors in the kidney. Am. J. Physiol. Renal Physiol. 291, F271–F281 (2006)

    ADS  CAS  Article  Google Scholar 

  17. Fernandez-Capetillo, O. et al. H2AX: the histone guardian of the genome. DNA Repair 3, 959–967 (2004)

    CAS  Article  Google Scholar 

  18. Rogakou, E. P. et al. Megabase chromatin domains involved in DNA double-strand breaks in vivo . J. Cell Biol. 146, 905–916 (1999)

    CAS  Article  Google Scholar 

  19. Berkovich, E., Monnat, R. J. & Kastan, M. B. Roles of ATM and NBS1 in chromatin structure modulation and DNA double-strand break repair. Nature Cell Biol. 9, 683–690 (2007)

    CAS  Article  Google Scholar 

  20. Berkovich, E., Monnat, R. J. & Kastan, M. B. Assessment of protein dynamics and DNA repair following generation of DNA double-strand breaks at defined genomic sites. Nature Protocols 3, 915–922 (2008)

    CAS  Article  Google Scholar 

  21. Matsuoka, S. et al. ATM and ATR substrate analysis reveals extensive protein networks responsive to DNA damage. Science 316, 1160–1166 (2007)

    ADS  CAS  Article  Google Scholar 

  22. Stokes, M. P. et al. Profiling of UV-induced ATM/ATR signaling pathways. Proc. Natl Acad. Sci. USA 104, 19855–19860 (2007)

    ADS  CAS  Article  Google Scholar 

  23. Lavin, M. F. & Kozlov, S. ATM activation and DNA damage response. Cell Cycle 6, 931–942 (2007)

    CAS  Article  Google Scholar 

  24. Stucki, M. et al. MDC1 directly binds phosphorylated histone H2AX to regulate cellular responses to DNA double-strand breaks. Cell 123, 1213–1226 (2005)

    CAS  Article  Google Scholar 

  25. Lee, M. S. et al. Structure of the BRCT repeat domain of MDC1 and its specificity for the free COOH-terminal end of the γ-H2AX histone tail. J. Biol. Chem. 280, 32053–32056 (2005)

    CAS  Article  Google Scholar 

  26. Kim, J. E., Minter-Dykhouse, K. & Chen, J. Signaling networks controlled by the MRN complex and MDC1 during early DNA damage responses. Mol. Carcinog. 45, 403–408 (2006)

    CAS  Article  Google Scholar 

  27. Wu, X. et al. ATM phosphorylation of Nijmegen breakage syndrome protein is required in a DNA damage response. Nature 405, 477–482 (2000)

    ADS  CAS  Article  Google Scholar 

  28. Celeste, A. et al. Histone H2AX phosphorylation is dispensable for the initial recognition of DNA breaks. Nature Cell Biol. 5, 675–679 (2003)

    CAS  Article  Google Scholar 

  29. Duilio, A. et al. A rat brain mRNA encoding a transcriptional activator homologous to the DNA binding domain of retroviral integrases. Nucleic Acids Res. 19, 5269–5274 (1991)

    CAS  Article  Google Scholar 

  30. Minopoli, G. et al. Essential roles for Fe65, Alzheimer amyloid precursor-binding protein, in the cellular response to DNA damage. J. Biol. Chem. 282, 831–835 (2007)

    CAS  Article  Google Scholar 

  31. Nakaya, T., Kawai, T. & Suzuki, T. Regulation of FE65 nuclear translocation and function by amyloid β-protein precursor in osmotically stressed cells. J. Biol. Chem. 283, 19119–19131 (2008)

    CAS  Article  Google Scholar 

  32. Celeste, A. et al. Genomic instability in mice lacking histone H2AX. Science 296, 922–927 (2002)

    ADS  CAS  Article  Google Scholar 

  33. Jemc, J. & Rebay, I. Identification of transcriptional targets of the dual-function transcription factor/phosphatase eyes absent. Dev. Biol. 310, 416–429 (2007)

    CAS  Article  Google Scholar 

  34. Xiao, A. et al. WSTF regulates the H2A.X DNA damage response via a novel tyrosine kinase activity. Nature 457, 57–62 (2009)

    ADS  CAS  Article  Google Scholar 

  35. Sambrook, J. & Russell, D. W. Molecular Cloning: a Laboratory Manual 3rd edn (Cold Spring Harbor Laboratory Press, 2001)

    Google Scholar 

Download references


We thank M. Kastan for providing reagents/technical assistance for the I-PpoI system. We thank V. Lunyak, J. Dixon and R. Koladner for review and discussions. We thank the laboratory of R. S. Johnson for use of equipment and advice on hypoxia incubations, as well as H. Taylor for animal care assistance and C. Nelson for cell culture assistance. We thank A. Nussenzweig, Y. Xu and H. Song for H2ax-/- MEFs. We thank J. Hightower and M. Fisher for assistance with figure and manuscript preparation. We additionally thank X. Li and W. Liu. M.G.R. is an HHMI Investigator. This work was supported by grants from NIH and NCI to M.G.R. and C.K.G. This work also was supported by the Sogang University Research Grant of 2008 to B.G.J and PCF and USAMRAA grants to M.G.R.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Michael G. Rosenfeld.

Supplementary information

Supplementary Information

This file contains Supplementary Methods, Supplementary Figures 1-12 with Legends and Supplementary Table 1. (PDF 497 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Cook, P., Ju, B., Telese, F. et al. Tyrosine dephosphorylation of H2AX modulates apoptosis and survival decisions. Nature 458, 591–596 (2009).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

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.


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