title: Cell cycle-dependent turnover of 5-hydroxymethyl cytosine in mouse embryonic stem cells.
abstract: Hydroxymethylcytosine in the genome is reported to be an intermediate of demethylation. In the present study, we demonstrated that maintenance methyltransferase Dnmt1 scarcely catalyzed hemi-hydroxymethylated DNA and that the hemi-hydroxymethylated DNA was not selectively recognized by the SRA domain of Uhrf1, indicating that hydroxymethylcytosine is diluted in a replication-dependent manner. A high level of 5-hydroxymethylcytosine in mouse embryonic stem cells was produced from the methylcytosine supplied mainly by de novo-type DNA methyltransferases Dnmt3a and Dnmt3b. The promoter regions of the HoxA gene cluster showed a high hydroxymethylation level whilst the methylcytosine level was quite low, suggesting that methylated CpG is actively hydroxylated during proliferation. All the results indicate that removal and production of hydroxymethylcytosine are regulated in replication-dependent manners in mouse embryonic stem cells.
authors: Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto, Japan.
authors: Junji
authors: J
authors: Otani
authors: Hironobu
authors: H
authors: Kimura
authors: Jafar
authors: J
authors: Sharif
authors: Takaho A
authors: TA
authors: Endo
authors: Yuichi
authors: Y
authors: Mishima
authors: Toru
authors: T
authors: Kawakami
authors: Haruhiko
authors: H
authors: Koseki
authors: Masahiro
authors: M
authors: Shirakawa
authors: Isao
authors: I
authors: Suetake
authors: Shoji
authors: S
authors: Tajima
mesh_terms: D044503:5-Methylcytosine / Q000031:analogs & derivatives; D000818:Animals; D022762:CCAAT-Enhancer-Binding Proteins; D002453:Cell Cycle; D049109:Cell Proliferation; D002469:Cell Separation; D018899:CpG Islands; D003596:Cytosine / Q000031:analogs & derivatives* / Q000737:chemistry; D004247:DNA / Q000032:analysis; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics; D019175:DNA Methylation; D000090205:DNA Methyltransferase 3A; D004261:DNA Replication; D053595:Embryonic Stem Cells / Q000166:cytology*; D005434:Flow Cytometry; D051379:Mice; D018345:Mice, Knockout; D005810:Multigene Family; D009687:Nuclear Proteins / Q000235:genetics; D011401:Promoter Regions, Genetic; D017434:Protein Structure, Tertiary; D044767:Ubiquitin-Protein Ligases; D000097584:DNA Methyltransferase 3B
chemical_list: D044503:5-Methylcytosine; D022762:CCAAT-Enhancer-Binding Proteins; D003596:Cytosine; D004247:DNA; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A; D009687:Nuclear Proteins; D044767:Ubiquitin-Protein Ligases; D000097584:DNA Methyltransferase 3B; C000715535:Dnmt3a protein, mouse; C011865:5-hydroxymethylcytosine; C000619381:Dnmt1 protein, mouse; C117054:Uhrf1 protein, mouse