Zscan4 Inhibits Maintenance DNA Methylation to Facilitate Telomere Elongation in Mouse Embryonic Stem Cells

Dan, Jiameng; Rousseau, Philippe; Hardikar, Swanand; Veland, Nicolás; Wong, Jiemin; Autexier, Chantal; Chen, Taiping

doi:10.1016/j.celrep.2017.07.070

Cited by 92 publications

(72 citation statements)

References 50 publications

(104 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Elevated DUX level in turn increases bipotent 2C-like cells in ESC culture through global transcriptional activation of 2C-specific genes, including genome caretaker genes, Zscan4 and Tcstv1/3 8,38 . In addition to their role in telomere elongation through activation of telomere sister chromatid exchange 7,8 , Zscan4 genes can also promote DNA repair by facilitating heterochromatin decondensation and DNA demethylation 6,15,39 . Therefore, ATR-mediated activation of Zscan4 genes likely contributes to ESCs genomic integrity in response to RS.…”

Section: Discussionmentioning

confidence: 99%

ATR expands embryonic stem cell fate potential in response to replication stress

Atashpaz

Shams

González

et al. 2020

Preprint

View full text Add to dashboard Cite

Unrepaired DNA damage during embryonic development can be potentially inherited by a large population of cells. However, the quality control mechanisms that minimize the contribution of damaged cells to developing embryos remain poorly understood.Here, we uncovered an ATR-and CHK1-mediated transcriptional response to replication stress (RS) in ESCs that induces genes expressed in totipotent two-cell (2C) stage embryos and 2C-like cells. This response is mediated by Dux, a multicopy retrogene defining the cleavage-specific transcriptional program in placental mammals. In response to RS, DUX triggers the transcription of 2C-like markers such as murine endogenous retrovirus-like elements (MERVL) and Zscan4. This response can also be elicited by ETAA1-mediated ATR activation in the absence of RS. ATR-mediated activation of DUX requires GSRF1 dependent post-transcriptional regulation of Dux mRNA. Strikingly, activation of ATR expands ESCs fate potential by extending their contribution to both embryonic and extraembryonic tissues. These findings define a novel ATR dependent pathway involved in maintaining genome stability in developing embryos by controlling ESCs fate in response to RS. BackgroundESCs are characterized by self-renewal and the ability to propagate for several cycles in vitro and in vivo 1 . Even if ESCs exhibit several markers of RS 2 , they are able to maintain genome integrity more efficiently than differentiated cells 1 . The mechanisms underlying such distinctive feature are largely unknown. ESC colonies harbor a small transient subpopulation of cells (2C-like cells) with functional andtranscriptional features of totipotent 2C-stage embryos 3-5 . Transition to 2C-like cells has been shown to promote maintenance of genome integrity and survival of ESCs in long-term culture 6-8 . In addition, several studies have demonstrated that transition to the 2C-like state confers expanded developmental potential to ESCs, making them capable of contributing to both embryonic and extra-embryonic tissues (also referred to as bidirectional cell fate potential) 3-5 . However, the molecular players underlying the transition to the 2C-like state in ESC culture and its possible physiological relevance in vivo in maintaining genome integrity and expanding cell fate potential in a developing embryo are not fully understood.Here we provide several lines of evidence that ATR and CHK1-mediated response to RS triggers the activation of 2C-specific genes in ESCs and mouse embryos. This transition is hampered in ESCs derived from ATR-deficient Seckel and CHK1 haploinsufficient mouse models and following ATR or CHK1 inhibition. Significantly, we show that ETAA1-mediated ATR activation is sufficient to trigger the formation of 2C-like cells in the absence of RS.Mechanistically, ATR-induced transition to 2C-like state is mediated by post-transcriptional regulation of the Dux gene, which shapes the transcriptional signature of 2C-like cells and totipotent 2C-stage embryos in placental mammals 9-11 . ATR-dependent regulation of Du...

show abstract

Section: Discussionmentioning

confidence: 99%

ATR expands embryonic stem cell fate potential in response to replication stress

Atashpaz

Shams

González

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…mESCs culture and generation of stable clones were described previously (Dan et al, 2017). Human cancer cell lines were cultured according to instructions of American Type Culture Collection.…”

Section: Methodsmentioning

confidence: 99%

“…Southern blot, dot blot and IF analyses of DNA methylation were performed as described previously (Dan et al, 2017). For bisulfite sequencing analysis, bisulfite conversion was performed using EZ DNA Methylation Kit (Zymo), PCR products cloned using NEB PCR cloning kit, and individual clones sequenced.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

The Arginine Methyltransferase PRMT6 Regulates DNA Methylation and Contributes to Global DNA Hypomethylation in Cancer

et al. 2017

Self Cite

View full text Add to dashboard Cite

SUMMARY DNA methylation plays crucial roles in chromatin structure and gene expression. Aberrant DNA methylation patterns, including global hypomethylation and regional hypermethylation, are associated with cancer and implicated in oncogenic events. How DNA methylation is regulated in developmental and cellular processes and dysregulated in cancer is poorly understood. Here, we show that PRMT6, a protein arginine methyltransferase responsible for asymmetric dimethylation of histone H3 arginine 2 (H3R2me2a), negatively regulates DNA methylation and that PRMT6 upregulation contributes to global DNA hypomethylation in cancer. Mechanistically, PRMT6 overexpression impairs chromatin association of UHRF1, an accessory factor of DNMT1, resulting in passive DNA demethylation. The effect is likely due to elevated H3R2me2a, which inhibits the interaction between UHRF1 and histone H3. Our work identifies a mechanistic link between protein arginine methylation and DNA methylation, which is disrupted in cancer.

show abstract

“…To circumvent this, we took advantage of a spontaneously occurring rare subpopulation of primed mouse embryonic stem cells (ESCs) that express transcripts usually restricted to ZGA, including the MERVL endogenous retrovirus and Zscan4 cluster (Macfarlan et al 2012;Zalzman et al 2010;Ishiuchi et al 2015;Eckersley-Maslin et al 2016). These "2C-like" ESCs also share several epigenetic features with the two-cell embryo, including global DNA hypomethylation (Eckersley-Maslin et al 2016;Dan et al 2017), decondensed chromatin (Eckersley-Maslin et al 2016;Ishiuchi et al 2015;Akiyama et al 2015) and increased histone mobility (Boskovic et al 2014).…”

Section: Candidate-based Screen For Epigenetic and Chromatin Regulatomentioning

confidence: 99%

Dppa2 and Dppa4 directly regulate the Dux driven zygotic transcriptional programme

Eckersley-Maslin

Alda-Catalinas

Blotenburg

et al. 2018

Preprint

View full text Add to dashboard Cite

The molecular regulation of zygotic genome activation (ZGA) in mammals remains poorly understood. Primed mouse embryonic stem cells contain a rare subset of "2C-like" cells that are epigenetically and transcriptionally similar to the two cell embryo and thus represent an ideal system for studying ZGA transcription regulation. Recently, the transcription factor Dux, expressed exclusively in the minor wave of ZGA, was described to activate many downstream ZGA transcripts. However, it remains unknown what upstream maternal factors initiate ZGA either in a Dux dependent or independent manner. Here we performed a candidate-based overexpression screen, identifying, amongst others, Developmental Pluripotency Associated 2 (Dppa2) and 4 (Dppa4) as positive regulators of 2C-like cells and ZGA transcription. In the germ line, promoter DNA demethylation coincides with upregulation of Dppa2 and Dppa4 which remain expressed until E7.5 when their promoters are remethylated. Furthermore, Dppa2 and Dppa4 are also expressed during iPSC reprogramming at the time 2C-like ZGA transcription transiently peaks. Through a combination of overexpression, knockdown, knockout and rescue experiments, together with transcriptional analyses, we show that Dppa2 and Dppa4 directly regulate the 2C-like cell population and associated transcripts, including Dux and the Zscan4 cluster. Importantly, we tease apart the molecular hierarchy in which the 2C-like transcriptional programme is initiated and stabilised. Dppa2 and Dppa4 require Dux to initiate 2C-like ZGA transcription, suggesting they act upstream by directly regulating Dux. Supporting this, ChIP-seq analysis revealed Dppa2 and Dppa4 bind to the Dux promoter and gene body and drive its expression. Zscan4c is also able to induce 2C-like cells in wild type cells, but, in contrast to Dux, can no longer do so in Dppa2/4 double knockout cells, suggesting it may act to stabilise rather than drive the transcriptional network. Our findings suggest a model in which Dppa2/4 binding to the Dux promoter leads to Dux upregulation and activation of the 2C-like transcriptional programme which is subsequently reinforced by Zscan4c.2

show abstract

Zscan4 Inhibits Maintenance DNA Methylation to Facilitate Telomere Elongation in Mouse Embryonic Stem Cells

Cited by 92 publications

References 50 publications

ATR expands embryonic stem cell fate potential in response to replication stress

ATR expands embryonic stem cell fate potential in response to replication stress

The Arginine Methyltransferase PRMT6 Regulates DNA Methylation and Contributes to Global DNA Hypomethylation in Cancer

Dppa2 and Dppa4 directly regulate the Dux driven zygotic transcriptional programme

Contact Info

Product

Resources

About