title: Inhibition of murine DNA methyltransferase Dnmt3a by DNA duplexes containing pyrimidine-2(1H)-one.
abstract: Here we studied the inhibition of the catalytic domain of Dnmt3a methyltransferase (Dnmt3a-CD) by DNA duplexes containing the mechanism-based inhibitor pyrimidine-2(1H)-one (P) instead of the target cytosine. It has been shown that conjugates of Dnmt3a-CD with P-DNA (DNA containing pyrimidine-2(1H)-one) are not stable to heating at 65°C in 0.1% SDS. The yield of covalent intermediate increases in the presence of the regulatory factor Dnmt3L. The importance of the DNA minor groove for covalent intermediate formation during the methylation reaction catalyzed by Dnmt3a-CD has been revealed. P-DNA was shown to inhibit Dnmt3a-CD; the IC(50) is 830 nM. The competitive mechanism of inhibition of Dnmt3a-CD by P-DNA has been elucidated. It is suggested that therapeutic effect of zebularine could be achieved by inhibition of not only Dnmt1 but also Dnmt3a.
authors.affiliation: Faculty of Chemistry, Lomonosov Moscow State University, Moscow, 119991, Russia. cherepanova@genebee.msu.ru
authors.forename: N A
authors.initials: NA
authors.lastname: Cherepanova
authors.forename: A L
authors.initials: AL
authors.lastname: Zhuze
authors.forename: E S
authors.initials: ES
authors.lastname: Gromova
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D020134:Catalytic Domain; D004247:DNA / Q000378:metabolism; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D019175:DNA Methylation; D000090205:DNA Methyltransferase 3A; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D051379:Mice; D011744:Pyrimidinones / Q000737:chemistry* / Q000494:pharmacology; D044366:Transition Temperature
chemical_list: C000715535:Dnmt3a protein, mouse; D004791:Enzyme Inhibitors; D011744:Pyrimidinones; D004247:DNA; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A