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The role of Tet3 DNA dioxygenase in epigenetic reprogramming by oocytes

Nature volume 477, pages 606–610 (2011)Cite this article

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Abstract

Sperm and eggs carry distinctive epigenetic modifications that are adjusted by reprogramming after fertilization1. The paternal genome in a zygote undergoes active DNA demethylation before the first mitosis2,3. The biological significance and mechanisms of this paternal epigenome remodelling have remained unclear4. Here we report that, within mouse zygotes, oxidation of 5-methylcytosine (5mC) occurs on the paternal genome, changing 5mC into 5-hydroxymethylcytosine (5hmC). Furthermore, we demonstrate that the dioxygenase Tet3 (ref. 5) is enriched specifically in the male pronucleus. In Tet3-deficient zygotes from conditional knockout mice, paternal-genome conversion of 5mC into 5hmC fails to occur and the level of 5mC remains constant. Deficiency of Tet3 also impedes the demethylation process of the paternal Oct4 and Nanog genes and delays the subsequent activation of a paternally derived Oct4 transgene in early embryos. Female mice depleted of Tet3 in the germ line show severely reduced fecundity and their heterozygous mutant offspring lacking maternal Tet3 suffer an increased incidence of developmental failure. Oocytes lacking Tet3 also seem to have a reduced ability to reprogram the injected nuclei from somatic cells. Therefore, Tet3-mediated DNA hydroxylation is involved in epigenetic reprogramming of the zygotic paternal DNA following natural fertilization and may also contribute to somatic cell nuclear reprogramming during animal cloning.

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Figure 1: Specific oxidation of methylcytosine and Tet3 distribution in the zygotic male pronucleus.
Figure 2: The role of Tet3 in 5mC oxidation, demethylation of paternal DNA, and activation of the paternal Oct4 allele.
Figure 3: Maternal Tet3 deficiency compromises embryonic development.
Figure 4: Tet3 contributes to reprogram the somatic nucleus transferred into oocytes.

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Acknowledgements

We thank C. Walsh and M. Rots for critical reading of the manuscript, J. Walter for discussions, H. Qi for providing cDNA of mouse oocytes, R. Zhang & Q. Cui for Tet3 cDNA, L. Li for help with 5hmCMP synthesis, Shanghai Research Center for Model Organisms for blastocyst injection, and J. Gao for mouse work. This study was supported by grants from the Ministry of Science and Technology China (2007CB947503 to G.-L.X., 2007CB947101 to J.L., and 2009CB941101 to G.-L.X. and J.L.), National Science Foundation of China (30730059 to G.-L.X. and 30871430 to J.L.), the Chinese Academy of Sciences (XDA01010301 to G.-L.X.; XDA01010403 and KSCX2-YW-R-110 to J.L.) and the NIH (GM078458 to Y.G.S.).

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Author notes

  1. Hai-Ping Wu

    Present address: Present address: Novartis Institutes for BioMedical Research Co., Shanghai 201203, China.,

  2. Tian-Peng Gu, Fan Guo and Hui Yang: These authors contributed equally to this work.

Authors and Affiliations

  1. Group of DNA Metabolism, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China

    Tian-Peng Gu, Fan Guo, Hai-Ping Wu, Gui-Fang Xu, Wei Liu, Zhi-Guo Xie & Guo-Liang Xu

  2. The State Key Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China

    Hui Yang, Linyu Shi & Jinsong Li

  3. The State Key Laboratory of Microbial Metabolism, School of Life Science and Biotechnology, Shanghai Jiaotong University, Shanghai, 200030, China

    Xinyi He & Zixin Deng

  4. Department of Cancer Biology, Beckman Research Institute of the City of Hope, Duarte, 91010, California, USA

    Seung-gi Jin & Gerd P. Pfeifer

  5. Department of Molecular and Cellular Biology, Beckman Research Institute of the City of Hope, Duarte, 91010, California, USA

    Khursheed Iqbal & Piroska E. Szabó

  6. Division of Endocrinology, Department of Medicine and BCMP, Diabetes, and Hypertension, Brigham and Women’s Hospital and Harvard Medical School, Boston, 02115, Massachusetts, USA

    Yujiang Geno Shi

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Contributions

G.-L.X. and J.L. conceived the projects. Y.G.S., H.-P.W. and G.-L.X. contributed to the knockout design. F.G., T.-P.G., H.-P.W., G.-F.X., and W.L. performed the experiments on early embryos. X.H. and Z.D. contributed to the synthesis of the 5hmC hapten. H.Y. and L.S. performed the nuclear transfer and embryo transfer experiments. S.-g.J., K.I., P.E.S., G.P.P. and Z.-G.X. characterized Tet3 expression in PGCs and ovaries. G.-L.X. wrote and G.P.P. revised the manuscript.

Corresponding authors

Correspondence to Jinsong Li or Guo-Liang Xu.

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Gu, TP., Guo, F., Yang, H. et al. The role of Tet3 DNA dioxygenase in epigenetic reprogramming by oocytes. Nature 477, 606–610 (2011). https://doi.org/10.1038/nature10443

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