scholarly journals Synergistic Function of DNA Methyltransferases Dnmt3a and Dnmt3b in the Methylation of Oct4 and Nanog

2007 ◽  
Vol 27 (24) ◽  
pp. 8748-8759 ◽  
Author(s):  
Jing-Yu Li ◽  
Min-Tie Pu ◽  
Ryutaro Hirasawa ◽  
Bin-Zhong Li ◽  
Yan-Nv Huang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Development ◽  
Gene Silencing ◽  
De Novo ◽  
Embryonic Stem ◽  
Dna Methyltransferases ◽  
Methylation Patterns ◽  
Genomic Methylation

ABSTRACT DNA methylation plays an important role in gene silencing in mammals. Two de novo methyltransferases, Dnmt3a and Dnmt3b, are required for the establishment of genomic methylation patterns in development. However, little is known about their coordinate function in the silencing of genes critical for embryonic development and how their activity is regulated. Here we show that Dnmt3a and Dnmt3b are the major components of a native complex purified from embryonic stem cells. The two enzymes directly interact and mutually stimulate each other both in vitro and in vivo. The stimulatory effect is independent of the catalytic activity of the enzyme. In differentiating embryonic carcinoma or embryonic stem cells and mouse postimplantation embryos, they function synergistically to methylate the promoters of the Oct4 and Nanog genes. Inadequate methylation caused by ablating Dnmt3a and Dnmt3b is associated with dysregulated expression of Oct4 and Nanog during the differentiation of pluripotent cells and mouse embryonic development. These results suggest that Dnmt3a and Dnmt3b form a complex through direct contact in living cells and cooperate in the methylation of the promoters of Oct4 and Nanog during cell differentiation. The physical and functional interaction between Dnmt3a and Dnmt3b represents a novel regulatory mechanism to ensure the proper establishment of genomic methylation patterns for gene silencing in development.

scholarly journals Establishment and Maintenance of Genomic Methylation Patterns in Mouse Embryonic Stem Cells by Dnmt3a and Dnmt3b

2003 ◽  
Vol 23 (16) ◽  
pp. 5594-5605 ◽  
Author(s):  
Taiping Chen ◽  
Yoshihide Ueda ◽  
Jonathan E. Dodge ◽  
Zhenjuan Wang ◽  
En Li
Keyword(s):  
Dna Methylation ◽  
De Novo ◽  
Es Cells ◽  
Embryonic Stem ◽  
Single Copy ◽  
Dna Methyltransferases ◽  
Methylation Patterns ◽  
Genomic Methylation ◽  
De Novo Methylation

ABSTRACT We have previously shown that the DNA methyltransferases Dnmt3a and Dnmt3b carry out de novo methylation of the mouse genome during early postimplantation development and of maternally imprinted genes in the oocyte. In the present study, we demonstrate that Dnmt3a and Dnmt3b are also essential for the stable inheritance, or “maintenance,” of DNA methylation patterns. Inactivation of both Dnmt3a and Dnmt3b in embryonic stem (ES) cells results in progressive loss of methylation in various repeats and single-copy genes. Interestingly, introduction of the Dnmt3a, Dnmt3a2, and Dnmt3b1 isoforms back into highly demethylated mutant ES cells restores genomic methylation patterns; these isoforms appear to have both common and distinct DNA targets, but they all fail to restore the maternal methylation imprints. In contrast, overexpression of Dnmt1 and Dnmt3b3 failed to restore DNA methylation patterns due to their inability to catalyze de novo methylation in vivo. We also show that hypermethylation of genomic DNA by Dnmt3a and Dnmt3b is necessary for ES cells to form teratomas in nude mice. These results indicate that genomic methylation patterns are determined partly through differential expression of different Dnmt3a and Dnmt3b isoforms.

scholarly journals A DNA signal from the Thy-1 gene defines de novo methylation patterns in embryonic stem cells.

1990 ◽  
Vol 10 (8) ◽  
pp. 4396-4400 ◽  
Author(s):  
M Szyf ◽  
G Tanigawa ◽  
P L McCarthy
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Dna Sequences ◽  
De Novo ◽  
Cpg Island ◽  
Embryonic Stem ◽  
Cpg Dinucleotides ◽  
Somatic Tissues ◽  
Methylation Patterns ◽  
De Novo Methylation

Although DNA can be extensively methylated de novo when introduced into pluripotent cells, the CpG island in the Thy-1 gene does not become methylated either in the mouse embryo or in embryonic stem cells. A 214-base-pair region near the promoter of the Thy-1 gene protects itself as well as heterologous DNA sequences from de novo methylation. We propose that this nucleotide sequence is representative of a class of important signals that limits de novo methylation in the embryo and establishes the pattern of hypomethylated CpG dinucleotides found in somatic tissues.

scholarly journals Derivation of sheep embryonic stem cells under optimized conditions

Reproduction ◽  
2020 ◽  
Vol 160 (5) ◽  
pp. 761-772 ◽  
Author(s):  
Marcela Vilarino ◽  
Delia Alba Soto ◽  
Yanina Soledad Bogliotti ◽  
Leqian Yu ◽  
Yanli Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
De Novo ◽  
Embryonic Stem ◽  
Culture Conditions ◽  
Livestock Species ◽  
Agricultural Applications ◽  
Wnt Inhibitor ◽  
Defined Culture

Until recently, it has been difficult to derive and maintain stable embryonic stem cells lines from livestock species. Sheep ESCs with characteristics similar to those described for rodents and primates have not been produced. We report the derivation of sheep ESCs under a chemically defined culture system containing fibroblast growth factor 2 (FGF2) and a tankyrase/Wnt inhibitor (IWR1). We also show that several culture conditions used for stabilizing naïve and intermediate pluripotency states in humans and mice were unsuitable to maintain ovine pluripotency in vitro. Sheep ESCs display a smooth dome-shaped colony morphology, and maintain an euploid karyotype and stable expression of pluripotency markers after more than 40 passages. We further demonstrate that IWR1 and FGF2 are essential for the maintenance of an undifferentiated state in de novo derived sheep ESCs. The derivation of stable pluripotent cell lines from sheep blastocysts represents a step forward toward understanding pluripotency regulation in livestock species and developing novel biomedical and agricultural applications.

scholarly journals Np95 interacts with de novo DNA methyltransferases, Dnmt3a and Dnmt3b, and mediates epigenetic silencing of the viral CMV promoter in embryonic stem cells

EMBO Reports ◽  
2009 ◽  
Vol 10 (11) ◽  
pp. 1259-1264 ◽  
Author(s):  
Daniela Meilinger ◽  
Karin Fellinger ◽  
Sebastian Bultmann ◽  
Ulrich Rothbauer ◽  
Ian Marc Bonapace ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
De Novo ◽  
Embryonic Stem ◽  
Dna Methyltransferases ◽  
Epigenetic Silencing ◽  
Cmv Promoter

A DNA signal from the Thy-1 gene defines de novo methylation patterns in embryonic stem cells

1990 ◽  
Vol 10 (8) ◽  
pp. 4396-4400
Author(s):  
M Szyf ◽  
G Tanigawa ◽  
P L McCarthy
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Dna Sequences ◽  
De Novo ◽  
Cpg Island ◽  
Embryonic Stem ◽  
Cpg Dinucleotides ◽  
Somatic Tissues ◽  
Methylation Patterns ◽  
De Novo Methylation

Although DNA can be extensively methylated de novo when introduced into pluripotent cells, the CpG island in the Thy-1 gene does not become methylated either in the mouse embryo or in embryonic stem cells. A 214-base-pair region near the promoter of the Thy-1 gene protects itself as well as heterologous DNA sequences from de novo methylation. We propose that this nucleotide sequence is representative of a class of important signals that limits de novo methylation in the embryo and establishes the pattern of hypomethylated CpG dinucleotides found in somatic tissues.

scholarly journals The distinct effects of MEK and GSK3 inhibition upon the methylome and transcriptome of mouse embryonic stem cells

2021 ◽  
Author(s):  
Julia Spindel ◽  
Christel Krueger ◽  
Felix Krueger ◽  
Evangelia K Papachristou ◽  
Kamal Kishore ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
De Novo ◽  
Embryonic Stem ◽  
Mek Inhibitor ◽  
Mouse Embryonic Stem Cells ◽  
Leukaemia Inhibitory Factor ◽  
Differentiation Markers ◽  
Chromatin Signature

Mouse embryonic stem cells (mESCs) were first cultured in vitro in serum-containing medium with leukaemia inhibitory factor, in which they exhibit heterogeneous expression of both pluripotency and some early differentiation markers. Over the last decade, however, it has become commonplace to grow mESCs with inhibitors of MEK and GSK3 signalling, which together elicit a more homogeneously 'naive' state of pluripotency. Whilst 2i/L-cultured mESCs have been shown to be globally hypomethylated, a comprehensive understanding of the distinct effects of these signalling inhibitors upon the DNA methylome is still lacking. Here we carried out whole genome bisulphite and RNA sequencing of mESCs grown with MEK or GSK3 inhibition alone, including different time points and concentrations of MEK inhibitor treatment. This demonstrated that MEK inhibition causes a dose-dependent impairment of maintenance methylation via loss of UHRF1 protein, as well as rapid impairment of de novo methylation. In contrast, GSK3 inhibition triggers impairment of de novo methylation alone, and consequent hypomethylation is enriched at enhancers with a 2i/L-specific chromatin signature and coincides with upregulation of nearby genes. Our study provides detailed insights into the epigenetic and transcriptional impacts of inhibiting MEK or GSK3 signalling in mouse pluripotent cells.

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