title: Mbd3/NURD complex regulates expression of 5-hydroxymethylcytosine marked genes in embryonic stem cells.
abstract: Numerous chromatin regulators are required for embryonic stem (ES) cell self-renewal and pluripotency, but few have been studied in detail. Here, we examine the roles of several chromatin regulators whose loss affects the pluripotent state of ES cells. We find that Mbd3 and Brg1 antagonistically regulate a common set of genes by regulating promoter nucleosome occupancy. Furthermore, both Mbd3 and Brg1 play key roles in the biology of 5-hydroxymethylcytosine (5hmC): Mbd3 colocalizes with Tet1 and 5hmC in vivo, Mbd3 knockdown preferentially affects expression of 5hmC-marked genes, Mbd3 localization is Tet1-dependent, and Mbd3 preferentially binds to 5hmC relative to 5-methylcytosine in vitro. Finally, both Mbd3 and Brg1 are themselves required for normal levels of 5hmC in vivo. Together, our results identify an effector for 5hmC, and reveal that control of gene expression by antagonistic chromatin regulators is a surprisingly common regulatory strategy in ES cells.
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605, USA.
authors.forename: Ozlem
authors.initials: O
authors.lastname: Yildirim
authors.forename: Ruowang
authors.initials: R
authors.lastname: Li
authors.forename: Jui-Hung
authors.initials: JH
authors.lastname: Hung
authors.forename: Poshen B
authors.initials: PB
authors.lastname: Chen
authors.forename: Xianjun
authors.initials: X
authors.lastname: Dong
authors.forename: Ly-Sha
authors.initials: LS
authors.lastname: Ee
authors.forename: Zhiping
authors.initials: Z
authors.lastname: Weng
authors.forename: Oliver J
authors.initials: OJ
authors.lastname: Rando
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Fazzio
mesh_terms: D044503:5-Methylcytosine / Q000031:analogs & derivatives; D000818:Animals; D042002:Chromatin Assembly and Disassembly; D003596:Cytosine / Q000031:analogs & derivatives* / Q000378:metabolism; D004265:DNA Helicases / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D006801:Humans; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex / Q000378:metabolism*; D051379:Mice; D009687:Nuclear Proteins / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D012319:RNA Polymerase II / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; C115369:MBD3 protein, human; C492013:Mbd3 protein, mouse; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; C540839:TET1 protein, mouse; D014157:Transcription Factors; C011865:5-hydroxymethylcytosine; D044503:5-Methylcytosine; D003596:Cytosine; D012319:RNA Polymerase II; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex; C513194:Smarca4 protein, mouse; D004265:DNA Helicases