Acetylation-induced transcription is required for active DNA demethylation in methylation-silenced genes.
A hallmark of vertebrate genes is that actively transcribed genes are hypomethylated in critical regulatory sequences. However, the mechanisms that link gene transcription and DNA hypomethylation are unclear. Using a trichostatin A (TSA)-induced replication-independent demethylation assay with HEK 293 cells, we show that RNA transcription is required for DNA demethylation. Histone acetylation precedes but is not sufficient to trigger DNA demethylation. Following histone acetylation, RNA polymerase II (RNAP II) interacts with the methylated promoter. Inhibition of RNAP II transcription with actinomycin D, alpha-amanitin, or CDK7-specific small interfering RNA inhibits DNA demethylation. H3 trimethyl lysine 4 methylation, a marker of actively transcribed genes, was associated with the cytomegalovirus promoter only after demethylation. TSA-induced demethylation of the endogenous cancer testis gene GAGE follows a similar sequence of events and is dependent on RNA transcription as well. These data suggest that DNA demethylation follows rather than precedes early transcription and point towards a novel function for DNA demethylation as a memory of actively transcribed genes.
D'Alessio
Ana C
AC
Department of Pharmacology and Therapeutics, McGill University, 3655 Promenade Sir William Osler, Montreal, Quebec H3G 1Y6, Canada.
Weaver
Ian C G
IC
Szyf
Moshe
M
D000107:Acetylation / Q000187:drug effects; D000951:Antigens, Neoplasm / Q000235:genetics; D002460:Cell Line; D018844:Cyclin-Dependent Kinases / Q000378:metabolism; D003587:Cytomegalovirus / Q000187:drug effects; D019175:DNA Methylation / Q000187:drug effects; D020868:Gene Silencing / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D008954:Models, Biological; D009363:Neoplasm Proteins / Q000235:genetics; D016366:Open Reading Frames / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012313:RNA / Q000235:genetics / Q000378:metabolism; D012319:RNA Polymerase II / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects; D000095043:Cyclin-Dependent Kinase-Activating Kinase
D000951:Antigens, Neoplasm; C095365:GAGE1 protein, human; D006657:Histones; D006877:Hydroxamic Acids; D009363:Neoplasm Proteins; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins; C012589:trichostatin A; D012313:RNA; D018844:Cyclin-Dependent Kinases; D012319:RNA Polymerase II; D000095043:Cyclin-Dependent Kinase-Activating Kinase
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