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Research article

Comparative analysis of DNA methylation patterns in transgenic Drosophila overexpressing mouse DNA methyltransferases

Cora MUND, Tanja MUSCH, Martin STRÖDICKE, Birte ASSMANN, En LI, Frank LYKO
Biochemical Journal Mar 15, 2004, 378 (3) 763-768; DOI: 10.1042/bj20031567
Cora MUND
Research Group Epigenetics, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 580, 69120 Heidelberg, Germany
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Tanja MUSCH
Research Group Epigenetics, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 580, 69120 Heidelberg, Germany
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Martin STRÖDICKE
Max-Planck-Institut für Molekulare Genetik, Ihnestrasse 73, 14195 Berlin, Germany
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Birte ASSMANN
Max-Planck-Institut für Molekulare Genetik, Ihnestrasse 73, 14195 Berlin, Germany
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En LI
Cardiovascular Research Center, Massachusetts General Hospital, 149 13th Street, Charlestown, MA 02129, U.S.A.
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Frank LYKO
Research Group Epigenetics, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 580, 69120 Heidelberg, Germany
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Abstract

DNA methyltransferases (Dnmts) mediate the epigenetic modification of eukaryotic genomes. Mammalian DNA methylation patterns are established and maintained by co-operative interactions among the Dnmt proteins Dnmt1, Dnmt3a and Dnmt3b. Owing to their simultaneous presence in mammalian cells, the activities of individual Dnmt have not yet been determined. This includes a fourth putative Dnmt, namely Dnmt2, which has failed to reveal any activity in previous assays. We have now established transgenic Drosophila strains that allow for individual overexpression of all known mouse Dnmts. Quantitative analysis of genomic cytosine methylation levels demonstrated a robust Dnmt activity for the de novo methyltransferases Dnmt3a and Dnmt3b. In addition, we also detected a weak but significant activity for Dnmt2. Subsequent methylation tract analysis by genomic bisulphite sequencing revealed that Dnmt3 enzymes preferentially methylated CpG dinucleotides in a processive manner, whereas Dnmt2 methylated isolated cytosine residues in a non-CpG dinucleotide context. Our results allow a direct comparison of the activities of mammalian Dnmts and suggest a significant functional specialization of these enzymes.

  • DNA methyltransferase
  • Dnmt1
  • Dnmt2
  • Dnmt3a
  • Dnmt3b
  • Drosophila

Footnotes

  • ↵1 Present address: Max-Delbrueck Center for Molecular Medicine, Robert-Roessle-Street 10, 13125 Berlin-Buch, Germany.

  • Abbreviations used: Dnmt, DNA methyltransferase; UAS, upstream activating sequence.

  • The Biochemical Society, London ©2004
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March 2004

Volume: 378 Issue: 3

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Comparative analysis of DNA methylation patterns in transgenic Drosophila overexpressing mouse DNA methyltransferases
Cora MUND, Tanja MUSCH, Martin STRÖDICKE, Birte ASSMANN, En LI, Frank LYKO
Biochemical Journal Mar 2004, 378 (3) 763-768; DOI: 10.1042/bj20031567
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Comparative analysis of DNA methylation patterns in transgenic Drosophila overexpressing mouse DNA methyltransferases
Cora MUND, Tanja MUSCH, Martin STRÖDICKE, Birte ASSMANN, En LI, Frank LYKO
Biochemical Journal Mar 2004, 378 (3) 763-768; DOI: 10.1042/bj20031567

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Keywords

DNA methyltransferase
Dnmt1
Dnmt2
Dnmt3a
Dnmt3b
Drosophila

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