Identification of DNMT1 selective antagonists using a novel scintillation proximity assay

Jessica A. Kilgore, Xinlin Du, Lisa Melito, Shuguang Wei, Changguang Wang, Hang Gyeong Chin, Bruce Posner, Sriharsa Pradhan, Joseph M. Ready, Noelle S. Williams

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

A novel scintillation proximity high throughput assay (SPA) to identify inhibitors of DNA methyltransferases was developed and used to screen over 180,000 compounds. The majority of the validated hits shared a quinone core and several were found to generate the reactive oxygen species, H2O 2. Inhibition of the production ofH2O2 by the addition of catalase blocked the ability of this group of compounds to inhibit DNA methyltransferase (DNMT) activity. However, a related compound, SW155246, was identified that existed in an already reduced form of the quinone. This compound did not generate H2O2, and catalase did not block its ability to inhibit DNA methyltransferase. SW155246 showed a 30-fold preference for inhibition of human DNMT1 versus human or murine DNMT3A or -3B, inhibited global methylation in HeLa cells, and reactivated expression of the tumor suppressor gene RASSF1A in A549 cells. To our knowledge, this work represents the first description of selective chemical inhibitors of the DNMT1 enzyme.

Original languageEnglish (US)
Pages (from-to)19673-19684
Number of pages12
JournalJournal of Biological Chemistry
Volume288
Issue number27
DOIs
StatePublished - Jul 5 2013

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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