Letter | Published:

The putative oncogene GASC1 demethylates tri- and dimethylated lysine 9 on histone H3

Nature volume 442, pages 307311 (20 July 2006) | Download Citation

Subjects

Abstract

Methylation of lysine and arginine residues on histone tails affects chromatin structure and gene transcription1,2,3. Tri- and dimethylation of lysine 9 on histone H3 (H3K9me3/me2) is required for the binding of the repressive protein HP1 and is associated with heterochromatin formation and transcriptional repression in a variety of species4,5,6. H3K9me3 has long been regarded as a ‘permanent’ epigenetic mark7,8. In a search for proteins and complexes interacting with H3K9me3, we identified the protein GASC1 (gene amplified in squamous cell carcinoma 1)9, which belongs to the JMJD2 (jumonji domain containing 2) subfamily of the jumonji family, and is also known as JMJD2C10. Here we show that three members of this subfamily of proteins demethylate H3K9me3/me2 in vitro through a hydroxylation reaction requiring iron and α-ketoglutarate as cofactors. Furthermore, we demonstrate that ectopic expression of GASC1 or other JMJD2 members markedly decreases H3K9me3/me2 levels, increases H3K9me1 levels, delocalizes HP1 and reduces heterochromatin in vivo. Previously, GASC1 was found to be amplified in several cell lines derived from oesophageal squamous carcinomas9,11,12, and in agreement with a contribution of GASC1 to tumour development, inhibition of GASC1 expression decreases cell proliferation. Thus, in addition to identifying GASC1 as a histone trimethyl demethylase, we suggest a model for how this enzyme might be involved in cancer development, and propose it as a target for anti-cancer therapy.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    , & Histone and chromatin cross-talk. Curr. Opin. Cell Biol. 15, 172–183 (2003)

  2. 2.

    , & The key to development: interpreting the histone code? Curr. Opin. Genet. Dev. 15, 163–176 (2005)

  3. 3.

    & Histones and histone modifications. Curr. Biol. 14, R546–R551 (2004)

  4. 4.

    et al. Selective recognition of methylated lysine 9 on histone H3 by the HP1 chromo domain. Nature 410, 120–124 (2001)

  5. 5.

    , , , & Methylation of histone H3 lysine 9 creates a binding site for HP1 proteins. Nature 410, 116–120 (2001)

  6. 6.

    & Heterochromatin: silence is golden. Curr. Biol. 13, R895–R898 (2003)

  7. 7.

    & A crack in histone lysine methylation. Cell 119, 903–906 (2004)

  8. 8.

    & Reversing histone methylation. Nature 436, 1103–1106 (2005)

  9. 9.

    et al. Identification of a novel gene, GASC1, within an amplicon at 9p23–24 frequently detected in esophageal cancer cell lines. Cancer Res. 60, 4735–4739 (2000)

  10. 10.

    & Identification and characterization of JMJD2 family genes in silico. Int. J. Oncol. 24, 1623–1628 (2004)

  11. 11.

    et al. A novel amplicon at 9p23-24 in squamous cell carcinoma of the esophagus that lies proximal to GASC1 and harbors NFIB. Jpn. J. Cancer Res. 92, 423–428 (2001)

  12. 12.

    et al. Genome-wide association study in esophageal cancer using GeneChip mapping 10K array. Cancer Res. 65, 2542–2546 (2005)

  13. 13.

    et al. Loss of the Suv39h histone methyltransferases impairs mammalian heterochromatin and genome stability. Cell 107, 323–337 (2001)

  14. 14.

    , & Methylation: lost in hydroxylation? EMBO Rep. 6, 315–320 (2005)

  15. 15.

    et al. Histone demethylation by a family of JmjC domain-containing proteins. Nature 439, 811–816 (2006)

  16. 16.

    et al. Structure of factor-inhibiting hypoxia-inducible factor (HIF) reveals mechanism of oxidative modification of HIF-1α. J. Biol. Chem. 278, 1802–1806 (2003)

  17. 17.

    , , , & Structure of human FIH-1 reveals a unique active site pocket and interaction sites for HIF-1 and von Hippel-Lindau. J. Biol. Chem. 278, 7558–7563 (2003)

  18. 18.

    & . The 2-His-1-carboxylate facial triad—an emerging structural motif in mononuclear non-heme iron(II) enzymes. Eur. J. Biochem. 250, 625–629 (1997)

  19. 19.

    , , & Effect of ascorbate on the activity of hypoxia-inducible factor in cancer cells. Cancer Res. 63, 1764–1768 (2003)

  20. 20.

    & Prolyl 4-hydroxylases and their protein disulfide isomerase subunit. Matrix Biol. 16, 357–368 (1998)

  21. 21.

    & The role of heterochromatin in centromere function. Phil. Trans. R. Soc. Lond. B 360, 569–579 (2005)

  22. 22.

    et al. Nucleosome binding by the bromodomain and PHD finger of the transcriptional cofactor p300. J. Mol. Biol. 337, 773–788 (2004)

  23. 23.

    et al. ONCOMINE: a cancer microarray database and integrated data-mining platform. Neoplasia 6, 1–6 (2004)

  24. 24.

    et al. Oncogene-induced senescence as an initial barrier in lymphoma development. Nature 436, 660–665 (2005)

  25. 25.

    et al. Crucial role of p53-dependent cellular senescence in suppression of Pten-deficient tumorigenesis. Nature 436, 725–730 (2005)

  26. 26.

    et al. Tumour biology: senescence in premalignant tumours. Nature 436, 642 (2005)

  27. 27.

    et al. BRAFE600-associated senescence-like cell cycle arrest of human naevi. Nature 436, 720–724 (2005)

  28. 28.

    et al. Regulation of LSD1 histone demethylase activity by its associated factors. Mol. Cell 19, 857–864 (2005)

  29. 29.

    et al. Gene expression profiling identifies clinically relevant subtypes of prostate cancer. Proc. Natl Acad. Sci. USA 101, 811–816 (2004)

  30. 30.

    et al. Gene expression alterations in prostate cancer predicting tumor aggression and preceding development of malignancy. J. Clin. Oncol. 22, 2790–2799 (2004)

Download references

Acknowledgements

We are grateful to U. Toftegaard and S. Keshtar for technical assistance. We thank A. Bracken, A. H. Lund and C. Storgaard Sørensen for critical reading of the manuscript. We thank members of the Helin laboratory and M. Salek for technical advice and support. This work was supported by grants from the Danish Cancer Society, the Novo Nordisk Foundation, the Danish Medical Research Council, the Danish Natural Science Research Council and the Danish Ministry of Science, Technology and Innovation. Author Contributions P.A.C.C. performed the in vitro binding experiments identifying GASC1 as an H3K9me3 interactor, suggested that GASC1 could be an H3K9me3-specific demethylase and co-wrote the paper. J.C. performed most DNA cloning steps and produced recombinant proteins. P.A.C.C. and J.C. established the in vitro demethylation assays, identified GASC1 as an H3K9me3-specific demethylase and performed various biochemical experiments. K.A. performed the in vivo experiments and immunofluorescence studies. K.H.H. designed the peptides and implemented the in vitro binding assays, which identified H3K9me3 interactors, and performed various biochemical experiments including preparation of samples for mass spectrometry. A.M. and J.R. performed mass spectrometry. T.A. performed in silico modelling studies. K.H. suggested the strategy to identify proteins binding to the modified histone tails, contributed to the planning of experiments and co-wrote the manuscript. All authors discussed the results and commented on the manuscript.

Author information

Author notes

    • Paul A. C. Cloos
    • , Jesper Christensen
    •  & Karl Agger

    *These authors contributed equally to this work

Affiliations

  1. Biotech Research & Innovation Centre, Fruebjergvej 3, 2100 Copenhagen, Denmark

    • Paul A. C. Cloos
    • , Jesper Christensen
    • , Karl Agger
    • , Torben Antal
    • , Klaus H. Hansen
    •  & Kristian Helin
  2. FIRC Institute of Molecular Oncology, Via Adamello 16, 20139 Milan, Italy

    • Alessio Maiolica
    •  & Juri Rappsilber
  3. Faculty of Health Sciences, University of Copenhagen, Blegdamsvej 3, 2200 Copenhagen, Denmark

    • Kristian Helin

Authors

  1. Search for Paul A. C. Cloos in:

  2. Search for Jesper Christensen in:

  3. Search for Karl Agger in:

  4. Search for Alessio Maiolica in:

  5. Search for Juri Rappsilber in:

  6. Search for Torben Antal in:

  7. Search for Klaus H. Hansen in:

  8. Search for Kristian Helin in:

Competing interests

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

Corresponding author

Correspondence to Kristian Helin.

Supplementary information

PDF files

  1. 1.

    Supplementary Notes

    This file contains the Supplementary Methods, additional references and Supplementary Figures 1–13.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nature04837

Further reading

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.