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© Research
Publication : Nucleic acids research

Development of a universal radioactive DNA methyltransferase inhibition test for high-throughput screening and mechanistic studies

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Nucleic acids research - 25 Aug 2013

Gros C, Chauvigné L, Poulet A, Menon Y, Ausseil F, Dufau I, Arimondo PB

Link to Pubmed [PMID] – 23980028

Nucleic Acids Res. 2013 Oct;41(19):e185

DNA methylation is an important epigenetic mark in eukaryotes, and aberrant pattern of this modification is involved in numerous diseases such as cancers. Interestingly, DNA methylation is reversible and thus is considered a promising therapeutic target. Therefore, there is a need for identifying new small inhibitors of C5 DNA methyltransferases (DNMTs). Despite the development of numerous in vitro DNMT assays, there is a lack of reliable tests suitable for high-throughput screening, which can also give insights into inhibitor mechanisms of action. We developed a new test based on scintillation proximity assay meeting these requirements. After optimizing our assay on human DNMT1 and calibrating it with two known inhibitors, we carried out S-Adenosyl-l-Methionine and DNA competition studies on three inhibitors and were able to determine each mechanism of action. Finally, we showed that our test was applicable to 3 other methyltransferases sources: human DNMT3A, bacterial M.SssI and cellular extracts as well.

https://www.ncbi.nlm.nih.gov/pubmed/23980028