Maintenance of self‐renewal ability of mouse embryonic stem cells in the absence of DNA methyltransferases Dnmt1, Dnmt3a and Dnmt3b

Tsumura, Akiko; Hayakawa, Tomohiro; Kumaki, Yuichi; Takebayashi, Shin‐ichiro; Sakaue, Masaki; Matsuoka, Chisa; Shimotohno, Kunitada; Ishikawa, Fuyuki; Li, En; Ueda, Hiroki R.; Nakayama, Jun‐ichi; Okano, Masaki

doi:10.1111/j.1365-2443.2006.00984.x

Cited by 497 publications

(453 citation statements)

References 44 publications

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“…Moreover, the primacy of TF binding relative to changes in DNA methylation has also been proposed during the Esrrb‐induced reprogramming of EpiSCs to a naïve state (Adachi et al , 2018). The ability of ESC self‐renewal to withstand complete loss of the DNA methylation machinery, with cell lethality of DNMT triple‐knockout ESCs only becoming apparent during differentiation (Tsumura et al , 2006), suggests that the increase in DNA methylation levels in committed cells reflects a change in the mode of gene regulation, from a dynamic process chiefly directed by TFs in pluripotent cells to one stabilized by DNA methylation (Festuccia et al , 2017). …”

Section: Discussionmentioning

confidence: 99%

Esrrb extinction triggers dismantling of naïve pluripotency and marks commitment to differentiation

et al. 2018

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Self‐renewal of embryonic stem cells (ESCs) cultured in LIF/fetal calf serum (FCS) is incomplete with some cells initiating differentiation. While this is reflected in heterogeneous expression of naive pluripotency transcription factors (TFs), the link between TF heterogeneity and differentiation is not fully understood. Here, we purify ESCs with distinct TF expression levels from LIF/FCS cultures to uncover early events during commitment from naïve pluripotency. ESCs carrying fluorescent Nanog and Esrrb reporters show Esrrb downregulation only in Nanoglow cells. Independent Esrrb reporter lines demonstrate that Esrrbnegative ESCs cannot effectively self‐renew. Upon Esrrb loss, pre‐implantation pluripotency gene expression collapses. ChIP‐Seq identifies different regulatory element classes that bind both OCT4 and NANOG in Esrrbpositive cells. Class I elements lose NANOG and OCT4 binding in Esrrbnegative ESCs and associate with genes expressed preferentially in naïve ESCs. In contrast, Class II elements retain OCT4 but not NANOG binding in ESRRB‐negative cells and associate with more broadly expressed genes. Therefore, mechanistic differences in TF function act cumulatively to restrict potency during exit from naïve pluripotency.

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Section: Discussionmentioning

confidence: 99%

Esrrb extinction triggers dismantling of naïve pluripotency and marks commitment to differentiation

et al. 2018

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“…We next determined the physiological importance of allosteric regulation of Dnmt3a by examining the impact of interface mutations on DNA methylation in ES cells. Stable cell lines expressing exogenous Dnmt3a at similar levels (Supplementary information, Figure S6A) were established from triple knockout (TKO) ES cells deficient in Dnmt1, Dnmt3a and Dnmt3b [24]. Cellular fractionation assay (Supplementary information, Data S1) confirmed that the mutant and wild-type proteins were similarly detected in the S1 and S2 fractions rich in euchromatin and heterochromatin, respectively (Supplementary information, Figure S6B).…”

Section: Mutations Affecting Allosteric Activation By Histone H3 Do Nmentioning

confidence: 91%

“…TKO (Dnmt31, Dnmt3a, Dnmt3b TKO [24]) stable ES cells lines ectopically expressing Dnmt3a were established by infection with lentivirus carrying a Dnmt3a expression cassette linked to the EGFP gene. ES clones expressing EGFP were picked and expanded.…”

Section: Establishment Of Stable Es Cell Lines and In Vitro Differentmentioning

confidence: 99%

Histone tails regulate DNA methylation by allosterically activating de novo methyltransferase

et al. 2011

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Cytosine methylation of genomic DNA controls gene expression and maintains genome stability. How a specific DNA sequence is targeted for methylation by a methyltransferase is largely unknown. Here, we show that histone H3 tails lacking lysine 4 (K4) methylation function as an allosteric activator for methyltransferase Dnmt3a by binding to its plant homeodomain (PHD). In vitro, histone H3 peptides stimulated the methylation activity of Dnmt3a up to 8-fold, in a manner reversely correlated with the level of K4 methylation. The biological significance of allosteric regulation was manifested by molecular modeling and identification of key residues in both the PHD and the catalytic domain of Dnmt3a whose mutations impaired the stimulation of methylation activity by H3 peptides but not the binding of H3 peptides. Significantly, these mutant Dnmt3a proteins were almost inactive in DNA methylation when expressed in mouse embryonic stem cells while their recruitment to genomic targets was unaltered. We therefore propose a two-step mechanism for de novo DNA methylation -first recruitment of the methyltransferase probably assisted by a chromatin-or DNA-binding factor, and then allosteric activation depending on the interaction between Dnmt3a and the histone tails -the latter might serve as a checkpoint for the methylation activity.

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“…Interestingly, ESCs from Dnmt1-deficeint mice or from Dnmt3a/Dnmt3b double knockout mice proliferate as undifferentiated stem cells [37], indicating that the self-renewal capability is preserved in ESCs with hypomethylated genomes. In addition, these mutant ESCs fail to undergo differentiation in vitro, possibly owing to a failure to repress pluripotency-related genes such as Oct4 and Nanog [38].…”

Section: Discussionmentioning

confidence: 99%

Promoter hypomethylation regulates CD133 expression in human gliomas

et al. 2008

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Maintenance of self‐renewal ability of mouse embryonic stem cells in the absence of DNA methyltransferases Dnmt1, Dnmt3a and Dnmt3b

Abstract: DNA methyltransferases

Cited by 497 publications

References 44 publications

Esrrb extinction triggers dismantling of naïve pluripotency and marks commitment to differentiation

Esrrb extinction triggers dismantling of naïve pluripotency and marks commitment to differentiation

Histone tails regulate DNA methylation by allosterically activating de novo methyltransferase

Promoter hypomethylation regulates CD133 expression in human gliomas

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