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A mammalian protein with specific demethylase activity for mCpG DNA

Nature volume 397, pages 579–583 (1999)Cite this article

Abstract

DNA-methylation patterns are important for regulating genome functions, and are determined by the enzymatic processes of methylation and demethylation. The demethylating enzyme has now been identified: a mammalian complementary DNA encodes a methyl-CpG-binding domain, bears a demethylase activity that transforms methylated cytosine bases to cytosine, and demethylates a plasmid when the cDNA is translated or transiently transfected into human embryonal kidney cells in vitro. The discovery of this DNA demethylase should provide a basis for the molecular and developmental analysis of the role of DNA methylation and demethylation.

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Figure 1: Human demethylase cDNA.
Figure 2: In vitro expression and demethylase activity of the human demethylase encoded by the cloned cDNA.
Figure 3: Expression of cloned demethylase cDNA in HEK cells.
Figure 4: DNA-demethylase activity purified from human cancer cells produces cytosine and exhibits no exonuclease or glycosylase activity.

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Acknowledgements

We thank B. ten Oever and H.-Y. Shiu for technical assistance and O. A. Mamer for help with mass spectrometry. This research was supported by the National Cancer Institute of Canada.

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Author notes

  1. Nadia Cervoni and Moshe Szyf: These authors contributed equally to this work.

  2. Moshe Szyf: Correspondence and requests for materials should be addressed to M.S. The DNA demethylase cDNA sequence has been deposited with Gen Bankunder accession number AF072242.

Authors and Affiliations

  1. Department of Pharmacology and Therapeutics, McGill University, 3655 Drummond Street, Montreal, H3G 1Y6, QC, Canada

    Sanjoy K. Bhattacharya & Shyam Ramchandani

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  1. Sanjoy K. Bhattacharya
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Bhattacharya, S., Ramchandani, S., Cervoni, N. et al. A mammalian protein with specific demethylase activity for mCpG DNA. Nature 397, 579–583 (1999). https://doi.org/10.1038/17533

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