De novo DNA methylation drives 5hmC accumulation in mouse zygotes

Rachel Amouroux; Buhe Nashun; Kenjiro Shirane; Shoma Nakagawa; Peter Ws Hill; Zelpha D'Souza; Manabu Nakayama; Masashi Matsuda; Aleksandra Turp; Elodie Ndjetehe; Vesela Encheva; Nobuaki R Kudo; Haruhiko Koseki; Hiroyuki Sasaki; Petra Hajkova

doi:10.1038/ncb3296

De novo DNA methylation drives 5hmC accumulation in mouse zygotes

Nat Cell Biol. 2016 Feb;18(2):225-233. doi: 10.1038/ncb3296. Epub 2016 Jan 11.

Authors

Affiliations

¹ Medical Research Council Clinical Sciences Centre, Imperial College London, London W12 0NN, United Kingdom.
² Division of Epigenomics and Development, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
³ Institute of Reproductive and Developmental Biology, Imperial College London, London W12 0NN, United Kingdom.
⁴ Chromosome Engineering Team, Department of Technology Development, Kazusa DNA Research Institute, 2-6-7 Kazusa-Kamatari, Kisarazu, Japan.
⁵ Laboratory for Developmental Genetics, RIKEN Center for Integrative Medical Sciences, 1-7-22 Suehiro, Tsurumi-ku, Yokohama, Japan.

^# Contributed equally.

PMID: 26751286
PMCID: PMC4765106
DOI: 10.1038/ncb3296

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.

Publication types

Letter

MeSH terms

5-Methylcytosine / metabolism*
Animals
Biomarkers / metabolism
Cellular Reprogramming*
Chromatography, Liquid
Cytosine / analogs & derivatives*
Cytosine / metabolism
DNA (Cytosine-5-)-Methyltransferase 1
DNA (Cytosine-5-)-Methyltransferases / genetics
DNA (Cytosine-5-)-Methyltransferases / metabolism
DNA Methylation*
DNA Methyltransferase 3A
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism
Dioxygenases
Embryo Culture Techniques
Epigenesis, Genetic*
Fertilization in Vitro
Fluorescent Antibody Technique
Gene Expression Regulation, Developmental
Kinetics
Mass Spectrometry
Mice
Mice, Knockout
Proto-Oncogene Proteins / genetics
Proto-Oncogene Proteins / metabolism
Zygote / metabolism*

Substances

Biomarkers
DNA-Binding Proteins
Dnmt3a protein, mouse
Proto-Oncogene Proteins
5-hydroxymethylcytosine
5-Methylcytosine
Cytosine
Dioxygenases
Tet3 protein, mouse
DNA (Cytosine-5-)-Methyltransferase 1
DNA (Cytosine-5-)-Methyltransferases
DNA Methyltransferase 3A
Dnmt1 protein, mouse

Abstract

Publication types

MeSH terms

Substances

Grants and funding