Epigenetics Protocols pp 157-177

Part of the Methods in Molecular Biology book series (MIMB, volume 791)

DNA Methyltransferase Assays

  • Renata Z. Jurkowska
  • Alexandre Ceccaldi
  • Yingying Zhang
  • Paola B. Arimondo
  • Albert Jeltsch
Protocol

Abstract

DNA methyltransferases are important enzymes and their inhibition has many potential applications. The investigation of DNA methyltransferases as well as screening for potential inhibitors requires specialized enzyme assays. In this chapter, we describe three DNA methyltransferase assays, each of them based on a different method: (1) An assay using radioactively labeled AdoMet and biotinylated DNA substrates that is ideal for enzymatic characterization of these enzymes. (2) An assay using bisulfite conversion of in vitro methylated DNA that is ideal to determine details of the methylation pattern introduced by DNA-(cytosine C5)-methyltransferases. (3) A novel fluorescence-coupled, restriction-based assay suitable for high-throughput screening of DNA methyltransferase inhibitors.

Key words

DNA methylation DNA methyltransferase Enzyme assay Bisulfite sequencing High-throughput screen Methyltransferase inhibitors 

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Renata Z. Jurkowska
    • 1
  • Alexandre Ceccaldi
    • 2
  • Yingying Zhang
    • 1
  • Paola B. Arimondo
    • 3
  • Albert Jeltsch
    • 1
  1. 1.Biochemistry Lab, School of Engineering and ScienceJacobs University BremenBremenGermany
  2. 2.Muséum National d’Histoire NaturelleParis CedexFrance
  3. 3.Muséum National d’Histoire NaturelleParisFrance

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