Meredith Gavin D, D'Ippolito Anthony, Dudas Miroslav, Zeidner Leigh C, Hostetter Logan, Faulds Kelsie, Arnold Thomas H, Popkie Anthony P, Doble Bradley W, Marnellos George, Adams Christopher, Wang Yulei, Phiel Christopher J
Thermo Fisher Scientific, Carlsbad, CA 92008.
Thermo Fisher Scientific, Carlsbad, CA 92008 Center for Human and Molecular Genetics, Nationwide Children's Hospital, Columbus, OH 43205.
Mol Biol Cell. 2015 Jun 1;26(11):2139-50. doi: 10.1091/mbc.E15-01-0013. Epub 2015 Apr 1.
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.
糖原合酶激酶-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Gsk-3在调控小鼠胚胎干细胞（ESC）印记基因座DNA甲基化方面的新作用，这表明由Gsk-3调控的表观遗传变化可能是Gsk-3信号传导中一个未被认识的方面。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（-/-）胚胎干细胞中进行下一代测序（MBD-Seq），将我们最初的观察扩展到整个小鼠基因组。与我们之前的数据一致，我们发现77%的已知印记基因座在Gsk-3缺陷的胚胎干细胞中DNA甲基化减少。更具体地说，我们明确鉴定了已被证实作为印记控制区域的区域内DNA甲基化的变化。在许多情况下，Gsk-3α（-/-）；Gsk-3β（-/-）胚胎干细胞中印记基因座处DNA甲基化的减少也伴随着基因表达的变化。此外，许多依赖Gsk-3的差异甲基化区域（DMR）与最近在单亲胚胎干细胞中鉴定出的DMR相同。我们的数据证明了Gsk-3活性在维持胚胎干细胞中大多数印记基因座的DNA甲基化方面的重要性，并强调了Gsk-3介导的信号转导在表观基因组中的重要性。
