title: De novo DNA methylation drives 5hmC accumulation in mouse zygotes.
abstract: Zygotic epigenetic reprogramming entails genome-wide DNA demethylation that is accompanied by Tet methylcytosine dioxygenase 3 (Tet3)-driven oxidation of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC; refs 1-4). Here we demonstrate using detailed immunofluorescence analysis and ultrasensitive LC-MS-based quantitative measurements that the initial loss of paternal 5mC does not require 5hmC formation. Small-molecule inhibition of Tet3 activity, as well as genetic ablation, impedes 5hmC accumulation in zygotes without affecting the early loss of paternal 5mC. Instead, 5hmC accumulation is dependent on the activity of zygotic Dnmt3a and Dnmt1, documenting a role for Tet3-driven hydroxylation in targeting de novo methylation activities present in the early embryo. Our data thus provide further insights into the dynamics of zygotic reprogramming, revealing an intricate interplay between DNA demethylation, de novo methylation and Tet3-driven hydroxylation.
authors.affiliation: Medical Research Council Clinical Sciences Centre, Imperial College London, London W12 0NN, United Kingdom.
authors.forename: Rachel
authors.initials: R
authors.lastname: Amouroux
authors.affiliation: Medical Research Council Clinical Sciences Centre, Imperial College London, London W12 0NN, United Kingdom.
authors.forename: Buhe
authors.initials: B
authors.lastname: Nashun
authors.affiliation: Division of Epigenomics and Development, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Shirane
authors.affiliation: Institute of Reproductive and Developmental Biology, Imperial College London, London W12 0NN, United Kingdom.
authors.forename: Shoma
authors.initials: S
authors.lastname: Nakagawa
authors.affiliation: Medical Research Council Clinical Sciences Centre, Imperial College London, London W12 0NN, United Kingdom.
authors.forename: Peter Ws
authors.initials: PW
authors.lastname: Hill
authors.affiliation: Medical Research Council Clinical Sciences Centre, Imperial College London, London W12 0NN, United Kingdom.
authors.forename: Zelpha
authors.initials: Z
authors.lastname: D'Souza
authors.affiliation: Chromosome Engineering Team, Department of Technology Development, Kazusa DNA Research Institute, 2-6-7 Kazusa-Kamatari, Kisarazu, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Nakayama
authors.affiliation: Laboratory for Developmental Genetics, RIKEN Center for Integrative Medical Sciences, 1-7-22 Suehiro, Tsurumi-ku, Yokohama, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Matsuda
authors.affiliation: Medical Research Council Clinical Sciences Centre, Imperial College London, London W12 0NN, United Kingdom.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Turp
authors.affiliation: Medical Research Council Clinical Sciences Centre, Imperial College London, London W12 0NN, United Kingdom.
authors.forename: Elodie
authors.initials: E
authors.lastname: Ndjetehe
authors.affiliation: Medical Research Council Clinical Sciences Centre, Imperial College London, London W12 0NN, United Kingdom.
authors.forename: Vesela
authors.initials: V
authors.lastname: Encheva
authors.affiliation: Institute of Reproductive and Developmental Biology, Imperial College London, London W12 0NN, United Kingdom.
authors.forename: Nobuaki R
authors.initials: NR
authors.lastname: Kudo
authors.affiliation: Laboratory for Developmental Genetics, RIKEN Center for Integrative Medical Sciences, 1-7-22 Suehiro, Tsurumi-ku, Yokohama, Japan.
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Koseki
authors.affiliation: Division of Epigenomics and Development, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Sasaki
authors.affiliation: Medical Research Council Clinical Sciences Centre, Imperial College London, London W12 0NN, United Kingdom.
authors.forename: Petra
authors.initials: P
authors.lastname: Hajkova
mesh_terms: D044503:5-Methylcytosine / Q000378:metabolism* / Q000031:analogs & derivatives; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D065150:Cellular Reprogramming; D002853:Chromatography, Liquid; D003596:Cytosine / Q000031:analogs & derivatives* / Q000378:metabolism; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D000090205:DNA Methyltransferase 3A; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D049308:Dioxygenases; D046149:Embryo Culture Techniques; D044127:Epigenesis, Genetic; D005307:Fertilization in Vitro; D005455:Fluorescent Antibody Technique; D018507:Gene Expression Regulation, Developmental; D007700:Kinetics; D013058:Mass Spectrometry; D051379:Mice; D018345:Mice, Knockout; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D015053:Zygote / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D004268:DNA-Binding Proteins; C000715535:Dnmt3a protein, mouse; D011518:Proto-Oncogene Proteins; C011865:5-hydroxymethylcytosine; D044503:5-Methylcytosine; D003596:Cytosine; D049308:Dioxygenases; C551915:Tet3 protein, mouse; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A; C000619381:Dnmt1 protein, mouse