title: Sirt1 Regulates DNA Methylation and Differentiation Potential of Embryonic Stem Cells by Antagonizing Dnmt3l.
abstract: Embryonic stem cell (ESC) abnormalities in genome methylation hamper the utility of their therapeutic derivatives; however, the underlying mechanisms are unknown. Here, we show that the nicotinamide adenine dinucleotide (NAD)-dependent deacetylase, Sirt1, selectively prevents abnormal DNA methylation of some developmental genes in murine ESCs by antagonizing Dnmt3l. Transcriptome and DNA methylome analyses demonstrated that Sirt1-null (Sirt1-/-) ESCs repress expression of a subset of imprinted and germline genes concomitant with increased DNA methylation of regulatory elements. Dnmt3l was highly expressed in Sirt1-/-ESCs, and knockdown partially rescued abnormal DNA methylation of the Sirt1 target genes. The Sirt1 protein suppressed transcription of Dnmt3l and physically interacted with the Dnmt3l protein, deacetylating and destabilizing Dnmt3l protein. Sirt1 deficiency delayed neurogenesis and spermatogenesis. These differentiation delays were significantly or partially abolished by reintroduction of Sirt1 cDNA or Dnmt3l knockdown. This study sheds light on mechanisms that restrain DNA methylation of developmentally vital genes operating in ESCs.
authors.affiliation: Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Korea; Department of Physiology, University of Ulsan College of Medicine, Seoul 05505, Korea.
authors.forename: Jinbeom
authors.initials: J
authors.lastname: Heo
authors.affiliation: Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Korea; Department of Physiology, University of Ulsan College of Medicine, Seoul 05505, Korea; Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul 03080, Korea.
authors.forename: Jisun
authors.initials: J
authors.lastname: Lim
authors.affiliation: Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Korea; Department of Physiology, University of Ulsan College of Medicine, Seoul 05505, Korea.
authors.forename: Seungun
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Korea; Department of Physiology, University of Ulsan College of Medicine, Seoul 05505, Korea.
authors.forename: Jaeho
authors.initials: J
authors.lastname: Jeong
authors.affiliation: Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Korea; Department of Physiology, University of Ulsan College of Medicine, Seoul 05505, Korea.
authors.forename: Hyunsook
authors.initials: H
authors.lastname: Kang
authors.affiliation: Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Korea; Department of Physiology, University of Ulsan College of Medicine, Seoul 05505, Korea.
authors.forename: YongHwan
authors.initials: Y
authors.lastname: Kim
authors.affiliation: Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Korea; Department of Physiology, University of Ulsan College of Medicine, Seoul 05505, Korea.
authors.forename: Jeong Wook
authors.initials: JW
authors.lastname: Kang
authors.affiliation: Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Korea.
authors.forename: Hwan Yeul
authors.initials: HY
authors.lastname: Yu
authors.affiliation: Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul 03080, Korea.
authors.forename: Eui Man
authors.initials: EM
authors.lastname: Jeong
authors.affiliation: Department of Convergence Medicine, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Korea; Clinical Proteomics Core Lab, Asan Institute for Life Sciences, Seoul 05505, Korea.
authors.forename: Kyunggon
authors.initials: K
authors.lastname: Kim
authors.affiliation: Stem Cell Institute at the James Graham Brown Cancer Center, University of Louisville, Louisville, KY 40202, USA.
authors.forename: Magda
authors.initials: M
authors.lastname: Kucia
authors.affiliation: Department of Medicine and Department of Pharmacology and Toxicology, Genomics Facility, University of Louisville, Louisville, KY 40202, USA.
authors.forename: Sabine J
authors.initials: SJ
authors.lastname: Waigel
authors.affiliation: Department of Medicine and Department of Pharmacology and Toxicology, Genomics Facility, University of Louisville, Louisville, KY 40202, USA.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Zacharias
authors.affiliation: Department of Medicine and Department of Pharmacology and Toxicology, Genomics Facility, University of Louisville, Louisville, KY 40202, USA.
authors.forename: Yinlu
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul 03080, Korea.
authors.forename: In-Gyu
authors.initials: IG
authors.lastname: Kim
authors.affiliation: Stem Cell Institute at the James Graham Brown Cancer Center, University of Louisville, Louisville, KY 40202, USA; Department of Regenerative Medicine, Warsaw Medical University, Warsaw 02-091, Poland. Electronic address: mzrata01@louisville.edu.
authors.forename: Mariusz Z
authors.initials: MZ
authors.lastname: Ratajczak
authors.affiliation: Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Korea; Department of Physiology, University of Ulsan College of Medicine, Seoul 05505, Korea. Electronic address: d0shin03@amc.seoul.kr.
authors.forename: Dong-Myung
authors.initials: DM
authors.lastname: Shin
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000037:antagonists & inhibitors*; D019175:DNA Methylation / Q000502:physiology*; D053595:Embryonic Stem Cells / Q000378:metabolism* / Q000502:physiology*; D005786:Gene Expression Regulation / Q000502:physiology; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009243:NAD / Q000378:metabolism; D055495:Neurogenesis / Q000502:physiology; D056564:Sirtuin 1 / Q000378:metabolism*; D013091:Spermatogenesis / Q000502:physiology; D014158:Transcription, Genetic / Q000502:physiology
chemical_list: D009243:NAD; C498796:Dnmt3l protein, mouse; D004248:DNA (Cytosine-5-)-Methyltransferases; C470545:Sirt1 protein, mouse; D056564:Sirtuin 1
keywords: DNA methylation; Dnmt3l; Sirt1; embryonic stem cell; epigenetic stability; genomic imprinting