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      The role and mechanisms of DNA methylation in the oocyte

      review-article
      ,
      Essays in Biochemistry
      Portland Press Ltd.
      chromatin, imprinting, methylation, oocytes

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          Abstract

          Epigenetic information in the mammalian oocyte has the potential to be transmitted to the next generation and influence gene expression; this occurs naturally in the case of imprinted genes. Therefore, it is important to understand how epigenetic information is patterned during oocyte development and growth. Here, we review the current state of knowledge of de novo DNA methylation mechanisms in the oocyte: how a distinctive gene-body methylation pattern is created, and the extent to which the DNA methylation machinery reads chromatin states. Recent epigenomic studies building on advances in ultra-low input chromatin profiling methods, coupled with genetic studies, have started to allow a detailed interrogation of the interplay between DNA methylation establishment and chromatin states; however, a full mechanistic description awaits.

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          DNMT3L connects unmethylated lysine 4 of histone H3 to de novo methylation of DNA.

          Mammals use DNA methylation for the heritable silencing of retrotransposons and imprinted genes and for the inactivation of the X chromosome in females. The establishment of patterns of DNA methylation during gametogenesis depends in part on DNMT3L, an enzymatically inactive regulatory factor that is related in sequence to the DNA methyltransferases DNMT3A and DNMT3B. The main proteins that interact in vivo with the product of an epitope-tagged allele of the endogenous Dnmt3L gene were identified by mass spectrometry as DNMT3A2, DNMT3B and the four core histones. Peptide interaction assays showed that DNMT3L specifically interacts with the extreme amino terminus of histone H3; this interaction was strongly inhibited by methylation at lysine 4 of histone H3 but was insensitive to modifications at other positions. Crystallographic studies of human DNMT3L showed that the protein has a carboxy-terminal methyltransferase-like domain and an N-terminal cysteine-rich domain. Cocrystallization of DNMT3L with the tail of histone H3 revealed that the tail bound to the cysteine-rich domain of DNMT3L, and substitution of key residues in the binding site eliminated the H3 tail-DNMT3L interaction. These data indicate that DNMT3L recognizes histone H3 tails that are unmethylated at lysine 4 and induces de novo DNA methylation by recruitment or activation of DNMT3A2.
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            Essential role for de novo DNA methyltransferase Dnmt3a in paternal and maternal imprinting.

            Imprinted genes are epigenetically marked during gametogenesis so that they are exclusively expressed from either the paternal or the maternal allele in offspring. Imprinting prevents parthenogenesis in mammals and is often disrupted in congenital malformation syndromes, tumours and cloned animals. Although de novo DNA methyltransferases of the Dnmt3 family are implicated in maternal imprinting, the lethality of Dnmt3a and Dnmt3b knockout mice has precluded further studies. We here report the disruption of Dnmt3a and Dnmt3b in germ cells, with their preservation in somatic cells, by conditional knockout technology. Offspring from Dnmt3a conditional mutant females die in utero and lack methylation and allele-specific expression at all maternally imprinted loci examined. Dnmt3a conditional mutant males show impaired spermatogenesis and lack methylation at two of three paternally imprinted loci examined in spermatogonia. By contrast, Dnmt3b conditional mutants and their offspring show no apparent phenotype. The phenotype of Dnmt3a conditional mutants is indistinguishable from that of Dnmt3L knockout mice, except for the discrepancy in methylation at one locus. These results indicate that both Dnmt3a and Dnmt3L are required for methylation of most imprinted loci in germ cells, but also suggest the involvement of other factors.
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              Targeted mutation of the DNA methyltransferase gene results in embryonic lethality

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                Author and article information

                Contributors
                Journal
                Essays Biochem
                Essays Biochem
                ebc
                Essays in Biochemistry
                Portland Press Ltd.
                0071-1365
                1744-1358
                December 2019
                29 November 2019
                : 63
                : 6 , DNA Methylation
                : 691-705
                Affiliations
                [1 ]Epigenetics Programme, Babraham Institute, Cambridge CB22 3AT, U.K.
                [2 ]Centre for Trophoblast Research, University of Cambridge, Cambridge CB2 3EG, U.K.
                Author notes
                Correspondence: Gintarė Sendžikaitė ( gintare.sendzikaite@ 123456babraham.ac.uk ) or Gavin Kelsey ( gavin.kelsey@ 123456babraham.ac.uk )
                Author information
                https://orcid.org/0000-0003-4569-3472
                http://orcid.org/0000-0002-9762-5634
                Article
                EBC20190043
                10.1042/EBC20190043
                6923320
                31782490
                78f6cb82-2064-48c0-a4ed-4ca25e4f3b9a
                © 2019 The Author(s).

                This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY).

                History
                : 17 September 2019
                : 29 October 2019
                : 29 October 2019
                Page count
                Pages: 15
                Categories
                DNA, Chromosomes & Chromosomal Structure
                Developmental Biology
                Gene Expression & Regulation
                Epigenetics
                Genomics
                Review Articles

                chromatin,imprinting,methylation,oocytes
                chromatin, imprinting, methylation, oocytes

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