title: Glycogen synthase kinase-3 (Gsk-3) plays a fundamental role in maintaining DNA methylation at imprinted loci in mouse embryonic stem cells.
abstract: 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.
authors.affiliation: Thermo Fisher Scientific, Carlsbad, CA 92008.
authors.forename: Gavin D
authors.initials: GD
authors.lastname: Meredith
authors.affiliation: Thermo Fisher Scientific, Carlsbad, CA 92008 Center for Human and Molecular Genetics, Nationwide Children's Hospital, Columbus, OH 43205.
authors.forename: Anthony
authors.initials: A
authors.lastname: D'Ippolito
authors.affiliation: Thermo Fisher Scientific, Carlsbad, CA 92008.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Dudas
authors.affiliation: Center for Human and Molecular Genetics, Nationwide Children's Hospital, Columbus, OH 43205.
authors.forename: Leigh C
authors.initials: LC
authors.lastname: Zeidner
authors.affiliation: Department of Integrative Biology, University of Colorado Denver, Denver, CO 80204.
authors.forename: Logan
authors.initials: L
authors.lastname: Hostetter
authors.affiliation: Department of Integrative Biology, University of Colorado Denver, Denver, CO 80204.
authors.forename: Kelsie
authors.initials: K
authors.lastname: Faulds
authors.affiliation: Department of Integrative Biology, University of Colorado Denver, Denver, CO 80204.
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Arnold
authors.affiliation: Graduate Program in Molecular, Cellular and Developmental Biology, Ohio State University, Columbus, OH 43210.
authors.forename: Anthony P
authors.initials: AP
authors.lastname: Popkie
authors.affiliation: McMaster Stem Cell and Cancer Research Institute, McMaster University, Hamilton, ON L8N 3Z5, Canada.
authors.forename: Bradley W
authors.initials: BW
authors.lastname: Doble
authors.affiliation: Thermo Fisher Scientific, Carlsbad, CA 92008.
authors.forename: George
authors.initials: G
authors.lastname: Marnellos
authors.affiliation: Thermo Fisher Scientific, Carlsbad, CA 92008.
authors.forename: Christopher
authors.initials: C
authors.lastname: Adams
authors.affiliation: Thermo Fisher Scientific, Foster City, CA 94404.
authors.forename: Yulei
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Center for Human and Molecular Genetics, Nationwide Children's Hospital, Columbus, OH 43205 Department of Integrative Biology, University of Colorado Denver, Denver, CO 80204 christopher.phiel@ucdenver.edu.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Phiel
mesh_terms: D000818:Animals; D019175:DNA Methylation; D053595:Embryonic Stem Cells / Q000378:metabolism*; D056426:Genetic Loci; D018392:Genomic Imprinting; D038362:Glycogen Synthase Kinase 3 / Q000235:genetics / Q000378:metabolism*; D000071679:Glycogen Synthase Kinase 3 beta; D059014:High-Throughput Nucleotide Sequencing; D051379:Mice; D018345:Mice, Knockout; D015398:Signal Transduction
chemical_list: C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; C000605507:Gsk3b protein, mouse; D038362:Glycogen Synthase Kinase 3; C448021:glycogen synthase kinase 3 alpha
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