Glycogen synthase kinase-3 (Gsk-3) plays a fundamental role in maintaining DNA methylation at imprinted loci in mouse embryonic stem cells.
Glycogen synthase kinase-3 (Gsk-3) plays a fundamental role in maintaining DNA methylation at imprinted loci in mouse embryonic stem cells.
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DOI：
10.1091/mbc.e15-01-0013
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发表时间：
2015-06-01
影响因子：
3.3
通讯作者：
Phiel CJ
中科院分区：
文献类型：
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作者：
Meredith GD;D'Ippolito A;Dudas M;Zeidner LC;Hostetter L;Faulds K;Arnold TH;Popkie AP;Doble BW;Marnellos G;Adams C;Wang Y;Phiel CJ
对Gsk-3α和Gsk-3β基因缺失的小鼠胚胎干细胞进行了全基因组DNA甲基化分析。将DNA甲基化模式与野生型细胞的DNA甲基化模式进行比较。超过75%的已知印迹基因座在Gsk-3敲除细胞中具有降低的DNA甲基化。糖原合成酶激酶-3（Glycogen synthase kinase-3，GSK-3）是多种信号转导途径的关键调节因子。最近，我们描述了一种新的作用，GSK-3在小鼠胚胎干细胞（ESCs）的印记位点的DNA甲基化的调节，这表明由GSK-3调节的表观遗传变化可能是一个未被认识的方面GSK-3信号。在这里，我们通过使用MethylMiner试剂盒富集甲基化DNA并在野生型和Gsk-3α−/−;Gsk-3β−/− ESC中进行下一代测序（MBD-Seq），将我们的初步观察扩展到整个小鼠基因组。与我们以前的数据一致，我们发现77%的已知印迹基因座在GSK-3缺陷的ESC中降低了DNA甲基化。更具体地说，我们明确地确定了已被证实作为印记控制区的区域内的DNA甲基化的变化。在许多情况下，Gsk-3α−/−;Gsk-3β−/− ESCs中印记位点的DNA甲基化降低也伴随着基因表达的变化。此外，许多GSK-3依赖的差异甲基化区域（DMR）与最近在单亲ESC中鉴定的DMR相同。我们的数据表明，GSK-3活性的重要性，在大多数胚胎干细胞的印记位点的DNA甲基化的维护和强调的重要性，GSK-3介导的信号转导的表观基因组。
A genome-wide analysis is given of DNA methylation in mouse embryonic stem cells in which both Gsk-3α and Gsk-3β have been genetically deleted. DNA methylation patterns are compared to those of wild-type cells. More than 75% of known imprinted loci have reduced DNA methylation in the Gsk-3–knockout cells. Glycogen synthase kinase-3 (Gsk-3) is a key regulator of multiple signal transduction pathways. Recently we described a novel role for Gsk-3 in the regulation of DNA methylation at imprinted loci in mouse embryonic stem cells (ESCs), suggesting that epigenetic changes regulated by Gsk-3 are likely an unrecognized facet of Gsk-3 signaling. Here we extend our initial observation to the entire mouse genome by enriching for methylated DNA with the MethylMiner kit and performing next-generation sequencing (MBD-Seq) in wild-type and Gsk-3α−/−;Gsk-3β−/− ESCs. Consistent with our previous data, we found that 77% of known imprinted loci have reduced DNA methylation in Gsk-3-deficient ESCs. More specifically, we unambiguously identified changes in DNA methylation within regions that have been confirmed to function as imprinting control regions. In many cases, the reduced DNA methylation at imprinted loci in Gsk-3α−/−;Gsk-3β−/− ESCs was accompanied by changes in gene expression as well. Furthermore, many of the Gsk-3–dependent, differentially methylated regions (DMRs) are identical to the DMRs recently identified in uniparental ESCs. Our data demonstrate the importance of Gsk-3 activity in the maintenance of DNA methylation at a majority of the imprinted loci in ESCs and emphasize the importance of Gsk-3–mediated signal transduction in the epigenome.