title: Gadd45a promotes DNA demethylation through TDG.
abstract: Growth arrest and DNA-damage-inducible protein 45 (Gadd45) family members have been implicated in DNA demethylation in vertebrates. However, it remained unclear how they contribute to the demethylation process. Here, we demonstrate that Gadd45a promotes active DNA demethylation through thymine DNA glycosylase (TDG) which has recently been shown to excise 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC) generated in Ten-eleven-translocation (Tet)-initiated oxidative demethylation. The connection of Gadd45a with oxidative demethylation is evidenced by the enhanced activation of a methylated reporter gene in HEK293T cells expressing Gadd45a in combination with catalytically active TDG and Tet. Gadd45a interacts with TDG physically and increases the removal of 5fC and 5caC from genomic and transfected plasmid DNA by TDG. Knockout of both Gadd45a and Gadd45b from mouse ES cells leads to hypermethylation of specific genomic loci most of which are also targets of TDG and show 5fC enrichment in TDG-deficient cells. These observations indicate that the demethylation effect of Gadd45a is mediated by TDG activity. This finding thus unites Gadd45a with the recently defined Tet-initiated demethylation pathway.
authors.affiliation: Group of DNA Metabolism, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Li
authors.affiliation: Group of DNA Metabolism, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Tian-Peng
authors.initials: TP
authors.lastname: Gu
authors.affiliation: Department of Biomedicine, University of Basel, Basel 4048, Switzerland.
authors.forename: Alain R
authors.initials: AR
authors.lastname: Weber
authors.affiliation: Group of DNA Metabolism, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Jia-Zhen
authors.initials: JZ
authors.lastname: Shen
authors.affiliation: Group of DNA Metabolism, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Bin-Zhong
authors.initials: BZ
authors.lastname: Li
authors.affiliation: Group of DNA Metabolism, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Zhi-Guo
authors.initials: ZG
authors.lastname: Xie
authors.affiliation: The State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
authors.forename: Ruichuan
authors.initials: R
authors.lastname: Yin
authors.affiliation: Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Guo
authors.affiliation: Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: Liu
authors.affiliation: Biodynamic Optical Imaging Center, College of Life Sciences, Peking University, Beijing 100871, China Ministry of Education Key Laboratory of Cell Proliferation and Differentiation, Peking University, Beijing 100871, China.
authors.forename: Fuchou
authors.initials: F
authors.lastname: Tang
authors.affiliation: The State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
authors.forename: Hailin
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Biomedicine, University of Basel, Basel 4048, Switzerland.
authors.forename: Primo
authors.initials: P
authors.lastname: Schär
authors.affiliation: Group of DNA Metabolism, The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China School of Life Science and Technology, ShanghaiTech University, 319 Yue Yang Road, Shanghai 200031, China glxu@sibcb.ac.cn.
authors.forename: Guo-Liang
authors.initials: GL
authors.lastname: Xu
mesh_terms: D000818:Animals; D018797:Cell Cycle Proteins / Q000235:genetics / Q000502:physiology*; D003596:Cytosine / Q000031:analogs & derivatives / Q000378:metabolism; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000378:metabolism; D049308:Dioxygenases; D053595:Embryonic Stem Cells / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D018345:Mice, Knockout; D009687:Nuclear Proteins / Q000235:genetics / Q000502:physiology*; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D044502:Thymine DNA Glycosylase / Q000378:metabolism*; D015533:Transcriptional Activation; D000097642:GADD45 Proteins
chemical_list: D018797:Cell Cycle Proteins; D003596:Cytosine; D004268:DNA-Binding Proteins; D049308:Dioxygenases; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; D044502:Thymine DNA Glycosylase; C560974:5-carboxylcytosine; C560973:5-formylcytosine; C499072:Gadd45a protein, mouse; D000097642:GADD45 Proteins; C541081:TET2 protein, human
references.citation: Nature. 2000 Feb 3;403(6769):501-2
references.pmid: 10676950
references.citation: Nat Protoc. 2013 Nov;8(11):2281-308
references.pmid: 24157548
references.citation: Mech Dev. 2002 Sep;117(1-2):15-23
references.pmid: 12204247
references.citation: Nat Genet. 2003 Mar;33 Suppl:245-54
references.pmid: 12610534
references.citation: Nat Cell Biol. 2004 Oct;6(10):984-90
references.pmid: 15448701
references.citation: Proc Natl Acad Sci U S A. 1988 Dec;85(23):8800-4
references.pmid: 3194391
references.citation: Annu Rev Biochem. 2005;74:481-514
references.pmid: 15952895
references.citation: Nat Methods. 2006 Feb;3(2):135-9
references.pmid: 16432524
references.citation: Curr Top Microbiol Immunol. 2006;301:283-315
references.pmid: 16570853
references.citation: Nature. 2007 Feb 8;445(7128):671-5
references.pmid: 17268471
references.citation: Epigenetics. 2006 Jul-Sep;1(3):127-30
references.pmid: 17965610
references.citation: Nature. 2008 Mar 6;452(7183):112-5
references.pmid: 18322535
references.citation: PLoS Genet. 2008 Mar;4(3):e1000013
references.pmid: 18369439
references.citation: Cell. 2008 Jun 27;133(7):1145-8
references.pmid: 18585349
references.citation: Cell. 2008 Dec 26;135(7):1201-12
references.pmid: 19109892
references.citation: Mol Cell. 2009 Feb 13;33(3):344-53
references.pmid: 19217408
references.citation: Science. 2009 Feb 20;323(5917):1074-7
references.pmid: 19119186
references.citation: PLoS Biol. 2009 Apr 28;7(4):e91
references.pmid: 19402749
references.citation: Science. 2009 May 15;324(5929):930-5
references.pmid: 19372391
references.citation: BMC Bioinformatics. 2010;11:230
references.pmid: 20459626
references.citation: Nature. 2011 Feb 17;470(7334):419-23
references.pmid: 21278727
references.citation: Nat Commun. 2011;2:241
references.pmid: 21407207
references.citation: Cell. 2011 Jul 8;146(1):67-79
references.pmid: 21722948
references.citation: Science. 2011 Sep 2;333(6047):1300-3
references.pmid: 21778364
references.citation: Science. 2011 Sep 2;333(6047):1303-7
references.pmid: 21817016
references.citation: Nature. 2011 Sep 29;477(7366):606-10
references.pmid: 21892189
references.citation: J Biol Chem. 2011 Oct 14;286(41):35334-8
references.pmid: 21862836
references.citation: Chromosoma. 2012 Feb;121(1):1-20
references.pmid: 22048164
references.citation: Trends Cell Biol. 2012 Apr;22(4):220-7
references.pmid: 22341196
references.citation: Development. 2012 Jun;139(11):1895-902
references.pmid: 22569552
references.citation: Cell. 2012 Jun 8;149(6):1368-80
references.pmid: 22608086
references.citation: Nucleic Acids Res. 2012 Jul;40(Web Server issue):W117-22
references.pmid: 22693217
references.citation: Philos Trans R Soc Lond B Biol Sci. 2013 Jan 5;368(1609):20110330
references.pmid: 23166394
references.citation: Cell. 2013 Apr 25;153(3):692-706
references.pmid: 23602152
references.citation: Cell. 2013 Apr 25;153(3):678-91
references.pmid: 23602153
references.citation: Nat Rev Mol Cell Biol. 2013 Jun;14(6):341-56
references.pmid: 23698584
references.citation: J Am Chem Soc. 2013 Jul 17;135(28):10396-403
references.pmid: 23768208
references.citation: Nature. 2013 Oct 24;502(7472):472-9
references.pmid: 24153300
references.citation: Cell. 2013 Dec 19;155(7):1545-55
references.pmid: 24315485
references.citation: Cell. 2014 Jan 16;156(1-2):45-68
references.pmid: 24439369
references.citation: Cell Stem Cell. 2014 Apr 3;14(4):512-22
references.pmid: 24529596
references.citation: Mol Cell. 2014 Aug 21;55(4):604-14
references.pmid: 25087872
references.citation: Cell Stem Cell. 2014 Oct 2;15(4):447-58
references.pmid: 25220291
references.citation: BMB Rep. 2014 Nov;47(11):609-18
references.pmid: 25341925
references.citation: Curr Biol. 2000 Apr 20;10(8):475-8
references.pmid: 10801417