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Planarian MBD2/3 is required for adult stem cell pluripotency independently of DNA methylation.

Jaber-Hijazi F, Lo PJ, Mihaylova Y, Foster JM, Benner JS, Tejada Romero B, Chen C, Malla S, Solana J, Ruzov A, Aziz Aboobaker A - Dev. Biol. (2013)

Bottom Line: Here we show the planarian methyl-CpG Binding Domain 2/3 (mbd2/3) gene is required for pASC differentiation during regeneration and tissue homeostasis.The genome does not have detectable levels of 5-methylcytosine (5(m)C) and we find no role for a potential DNA methylase.We conclude that MBD proteins may have had an ancient role in broadly controlling animal stem cell pluripotency, but that DNA methylation is not involved in planarian stem cell differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Zoology, Tinbergen Building, South Parks Road, University of Oxford, Oxford OX1 3PS, United Kingdom.

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Lack of evidence for a role for DNA methylation in pASC biology Smed-dnmt-2 is expressed in the germ line and germ line stem cells (A) Scale bars, 500 µm. gDNA digestion with the methylation dependent restriction enzyme McrBC does not indicate any DNA cytosine methylation in the Schmidtea mediterranea genome as compared with that of humans. −: no enzyme; +: with enzyme (B). Control chromatogram after HPLC-MS displaying the sensitivity of nucleosides detection, in particular for 5mC. (C) A representative chromatogram of S. mediterranea nucleoside analysis indicating the lack of 5mC (D).
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f0035: Lack of evidence for a role for DNA methylation in pASC biology Smed-dnmt-2 is expressed in the germ line and germ line stem cells (A) Scale bars, 500 µm. gDNA digestion with the methylation dependent restriction enzyme McrBC does not indicate any DNA cytosine methylation in the Schmidtea mediterranea genome as compared with that of humans. −: no enzyme; +: with enzyme (B). Control chromatogram after HPLC-MS displaying the sensitivity of nucleosides detection, in particular for 5mC. (C) A representative chromatogram of S. mediterranea nucleoside analysis indicating the lack of 5mC (D).

Mentions: Some MBD proteins are thought to act through binding 5mC and affect gene expression (Hendrich and Bird, 1998; Jaenisch and Bird, 2003). We looked for evidence for the presence 5mC in genomic DNA (gDNA) and for genes encoding potential DNA methyltransferases. We identified a single gene with identity to known DNA methyltransferases, Smed-dnmt2 (Supplementary Fig. 5A and B). Neither the observed expression, which was confined to the germline (Wang et al., 2007) (Fig. 7A), nor RNAi experiments indicated any role in regeneration (Supplementary Fig. 5C and D). Another recent study reported a DMNT1 gene in S. mediterranea (Zeng et al., 2013), but we have found only a DMNT2 ortholog in line with a recent study that only found DMNT2 encoding genes across the platyhelminthes (Geyer et al., 2013).


Planarian MBD2/3 is required for adult stem cell pluripotency independently of DNA methylation.

Jaber-Hijazi F, Lo PJ, Mihaylova Y, Foster JM, Benner JS, Tejada Romero B, Chen C, Malla S, Solana J, Ruzov A, Aziz Aboobaker A - Dev. Biol. (2013)

Lack of evidence for a role for DNA methylation in pASC biology Smed-dnmt-2 is expressed in the germ line and germ line stem cells (A) Scale bars, 500 µm. gDNA digestion with the methylation dependent restriction enzyme McrBC does not indicate any DNA cytosine methylation in the Schmidtea mediterranea genome as compared with that of humans. −: no enzyme; +: with enzyme (B). Control chromatogram after HPLC-MS displaying the sensitivity of nucleosides detection, in particular for 5mC. (C) A representative chromatogram of S. mediterranea nucleoside analysis indicating the lack of 5mC (D).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3824064&req=5

f0035: Lack of evidence for a role for DNA methylation in pASC biology Smed-dnmt-2 is expressed in the germ line and germ line stem cells (A) Scale bars, 500 µm. gDNA digestion with the methylation dependent restriction enzyme McrBC does not indicate any DNA cytosine methylation in the Schmidtea mediterranea genome as compared with that of humans. −: no enzyme; +: with enzyme (B). Control chromatogram after HPLC-MS displaying the sensitivity of nucleosides detection, in particular for 5mC. (C) A representative chromatogram of S. mediterranea nucleoside analysis indicating the lack of 5mC (D).
Mentions: Some MBD proteins are thought to act through binding 5mC and affect gene expression (Hendrich and Bird, 1998; Jaenisch and Bird, 2003). We looked for evidence for the presence 5mC in genomic DNA (gDNA) and for genes encoding potential DNA methyltransferases. We identified a single gene with identity to known DNA methyltransferases, Smed-dnmt2 (Supplementary Fig. 5A and B). Neither the observed expression, which was confined to the germline (Wang et al., 2007) (Fig. 7A), nor RNAi experiments indicated any role in regeneration (Supplementary Fig. 5C and D). Another recent study reported a DMNT1 gene in S. mediterranea (Zeng et al., 2013), but we have found only a DMNT2 ortholog in line with a recent study that only found DMNT2 encoding genes across the platyhelminthes (Geyer et al., 2013).

Bottom Line: Here we show the planarian methyl-CpG Binding Domain 2/3 (mbd2/3) gene is required for pASC differentiation during regeneration and tissue homeostasis.The genome does not have detectable levels of 5-methylcytosine (5(m)C) and we find no role for a potential DNA methylase.We conclude that MBD proteins may have had an ancient role in broadly controlling animal stem cell pluripotency, but that DNA methylation is not involved in planarian stem cell differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Zoology, Tinbergen Building, South Parks Road, University of Oxford, Oxford OX1 3PS, United Kingdom.

Show MeSH