De novo DNA cytosine methyltransferase activities in mouse embryonic stem cells
Abstract
It has been a controversial issue as to how many DNA cytosine methyltransferase mammalian cells have and whether de novo methylation and maintenance methylation activities are encoded by a single gene or two different genes. To address these questions, we have generated a null mutation of the only known mammalian DNA methyl-transferase gene through homologous recombination in mouse embryonic stem cells and found that the development of the homozygous embryos is arrested prior to the 8-somite stage. Surprisingly, the null mutant embryonic stem cells are viable and contain low but stable levels of methyl cytosine and methyltransferase activity, suggesting the existence of a second DNA methyltransferase in mammalian cells. Further studies indicate that de novo methylation activity is not impaired by the mutation as integrated provirus DNA in MoMuLV-infected homozygous embryonic stem cells become methylated at a similar rate as in wild-type cells. Differentiation of mutant cells results in further reduction of methyl cytosine levels, consistent with the de novo methylation activity being down regulated in differentiated cells. These results provide the first evidence that an independently encoded DNA methyl-transferase is present in mammalian cells which is capable of de novo methylating cellular and viral DNA in vivo.
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Global deletion of mouse Dnmt1, Dnmt3b, or both Dnmt3a + Dnmt3b results in midgestation lethality (GD 9.5-11.5), while deletion of only Dnmt3a produces severe growth retardation and lethality by 4 weeks of age.
“Global deletion of mouse Dnmt1, Dnmt3b, or both Dnmt3a + Dnmt3b results in midgestation lethality [gestational day (GD) 9.5–11.5], while deletion of only Dnmt3a produces severe growth retardation and lethality by 4 weeks of age [ 70–72 ].”
From DNA methylation: a mechanism linking environmental chemical exposures to risk of autism spectrum disorders? (2016) · cites this paper as [71]
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- Authors
- Lei, H., Oh, S. P., Okano, M., Jüttermann, R., Goss, K. A., Jaenisch, R., & Li, E.
- Affiliations
- Massachusetts General Hospital (US)
- Harvard University (US)
- Whitehead Institute for Biomedical Research (US)
- Venue
- Development
- Publisher
- The Company of Biologists
- Published
- 1996-10-01
- Volume
- 122
- Issue
- 10
- Pages
- 3195-3205
- ISSN
- 0950-1991 (ISSN-L)
- 1477-9129
- OpenAlex
- W2157896232
- Cited by
- 816
- References
- 44
- Citations by year
- 2026: 11 · 2025: 11 · 2024: 13 · 2023: 9 · 2022: 21 · 2021: 17 · 2020: 27 · 2019: 22 · 2018: 23 · 2017: 28 · 2016: 39 · 2015: 32 · 2014: 36 · 2013: 28 · 2012: 25
- FWCI
- 10.69
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Development (publishedVersion)
- PubMed (publishedVersion)
- HighWire Press Open Archive (submittedVersion)
- Funders
- National Institutes of Health (5r35ca044339-13, 2r01gm052106-06)
- National Cancer Institute (R-35-CA44339)
- National Institute of General Medical Sciences (GM52106)
- Eunice Kennedy Shriver National Institute of Child Health and Human Development (F32HD007945, F32 HD07945-01)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic Syndromes and Imprinting
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Keywords
- DNA methyltransferases, de novo methylation, maintenance methylation, DNMT1, DNMT3, mouse embryonic stem cells, gene knockout, cell differentiation, epigenetic inheritance
- MeSH terms
- Animals, Cell Differentiation, Cell Line, Cell Survival, DNA, Viral, Female, Moloney murine leukemia virus, Pregnancy, Proviruses, DNA-Cytosine Methylases, Gene Deletion, DNA Methylation, Embryonic Development, Mice
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