Vitamin C induces Tet-dependent DNA demethylation and a blastocyst-like state in ES cells
Top Cited Papers
Open Access
- 30 June 2013
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature
- Vol. 500 (7461), 222-226
- https://doi.org/10.1038/nature12362
Abstract
Vitamin C is a direct regulator of Tet enzyme activity and DNA methylation fidelity in mouse ES cells; addition of vitamin C promotes Tet activity, increases 5-hydroxymethlycytosine (5hmC) and DNA demethylation of many gene promoters, upregulates demethylated germline genes, and induces a state that more closely approximates that of the inner cell mass of the blastocyst. The Tet enzymes regulate DNA methylation by converting 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) and other oxidized variants. Here vitamin C, a cofactor of enzymes of the same family as the Tets, is shown to be a direct regulator of Tet activity in mouse embryonic stem cells, which are typically cultured in the absence of vitamin C. Addition of vitamin C to the culture medium leads to increased 5hmC content and the demethylation of numerous gene promoters. The remodelled DNA methylation and gene expression patterns resemble the DNA demethylation that occurs in the inner cell mass of the early embryo. DNA methylation is a heritable epigenetic modification involved in gene silencing, imprinting, and the suppression of retrotransposons1. Global DNA demethylation occurs in the early embryo and the germ line2,3, and may be mediated by Tet (ten eleven translocation) enzymes4,5,6, which convert 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC)7. Tet enzymes have been studied extensively in mouse embryonic stem (ES) cells8,9,10,11,12, which are generally cultured in the absence of vitamin C, a potential cofactor for Fe(ii) 2-oxoglutarate dioxygenase enzymes such as Tet enzymes. Here we report that addition of vitamin C to mouse ES cells promotes Tet activity, leading to a rapid and global increase in 5hmC. This is followed by DNA demethylation of many gene promoters and upregulation of demethylated germline genes. Tet1 binding is enriched near the transcription start site of genes affected by vitamin C treatment. Importantly, vitamin C, but not other antioxidants, enhances the activity of recombinant Tet1 in a biochemical assay, and the vitamin-C-induced changes in 5hmC and 5mC are entirely suppressed in Tet1 and Tet2 double knockout ES cells. Vitamin C has a stronger effect on regions that gain methylation in cultured ES cells compared to blastocysts, and in vivo are methylated only after implantation. In contrast, imprinted regions and intracisternal A particle retroelements, which are resistant to demethylation in the early embryo2,13, are resistant to vitamin-C-induced DNA demethylation. Collectively, the results of this study establish vitamin C as a direct regulator of Tet activity and DNA methylation fidelity in ES cells.Keywords
This publication has 42 references indexed in Scilit:
- The Dynamics of Genome-wide DNA Methylation Reprogramming in Mouse Primordial Germ CellsMolecular Cell, 2012
- Tet1 controls meiosis by regulating meiotic gene expressionNature, 2012
- The Transcriptional and Epigenomic Foundations of Ground State PluripotencyCell, 2012
- Ascorbic acid prevents loss of Dlk1-Dio3 imprinting and facilitates generation of all–iPS cell mice from terminally differentiated B cellsNature Genetics, 2012
- Tet1 Is Dispensable for Maintaining Pluripotency and Its Loss Is Compatible with Embryonic and Postnatal DevelopmentCell Stem Cell, 2011
- Dual functions of Tet1 in transcriptional regulation in mouse embryonic stem cellsNature, 2011
- Tet1 and Tet2 Regulate 5-Hydroxymethylcytosine Production and Cell Lineage Specification in Mouse Embryonic Stem CellsCell Stem Cell, 2011
- Tracing the Derivation of Embryonic Stem Cells from the Inner Cell Mass by Single-Cell RNA-Seq AnalysisCell Stem Cell, 2010
- The ground state of embryonic stem cell self-renewalNature, 2008
- Promoter CpG Methylation Contributes to ES Cell Gene Regulation in Parallel with Oct4/Nanog, PcG Complex, and Histone H3 K4/K27 TrimethylationCell Stem Cell, 2008