Home LiteratureArticle Details
PMID: 17938196 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Synergistic function of DNA methyltransferases Dnmt3a and Dnmt3b in the methylation of Oct4 and Nanog.

Molecular and cellular biology ·Vol. 27 ·No. 24 ·2007-12-00 ·Pages 8748-59

Li JY, Pu MT, Hirasawa R, Li BZ, Huang YN, Zeng R, Jing NH, Chen T, Li E, Sasaki H, Xu GL

Abstract

DNA methylation plays an important role in gene silencing in mammals. Two de novo methyltransferases, Dnmt3a and Dnmt3b, are required for the establishment of genomic methylation patterns in development. However, little is known about their coordinate function in the silencing of genes critical for embryonic development and how their activity is regulated. Here we show that Dnmt3a and Dnmt3b are the major components of a native complex purified from embryonic stem cells. The two enzymes directly interact and mutually stimulate each other both in vitro and in vivo. The stimulatory effect is independent of the catalytic activity of the enzyme. In differentiating embryonic carcinoma or embryonic stem cells and mouse postimplantation embryos, they function synergistically to methylate the promoters of the Oct4 and Nanog genes. Inadequate methylation caused by ablating Dnmt3a and Dnmt3b is associated with dysregulated expression of Oct4 and Nanog during the differentiation of pluripotent cells and mouse embryonic development. These results suggest that Dnmt3a and Dnmt3b form a complex through direct contact in living cells and cooperate in the methylation of the promoters of Oct4 and Nanog during cell differentiation. The physical and functional interaction between Dnmt3a and Dnmt3b represents a novel regulatory mechanism to ensure the proper establishment of genomic methylation patterns for gene silencing in development.

MeSH Terms
Animals Cell Differentiation/drug effects Cell Line, Tumor Chromatography, Affinity DNA (Cytosine-5-)-Methyltransferases/deficiency,isolation & purification,metabolism DNA Methylation/drug effects DNA Methyltransferase 3A DNA-Binding Proteins/genetics Embryonic Development/drug effects Embryonic Stem Cells/cytology,drug effects,enzymology,metabolism Homeodomain Proteins/genetics Mice Nanog Homeobox Protein Octamer Transcription Factor-3/genetics Promoter Regions, Genetic/genetics Protein Binding/drug effects Tretinoin/pharmacology
Chemicals
DNA-Binding Proteins Dnmt3a protein, mouse Homeodomain Proteins Nanog Homeobox Protein Nanog protein, mouse Octamer Transcription Factor-3 Pou5f1 protein, mouse Tretinoin DNA (Cytosine-5-)-Methyltransferases DNA Methyltransferase 3A DNA methyltransferase 3B
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Li Jing-Yu
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai 200031, People's Republic of China.
Pu Min-Tie
Hirasawa Ryutaro
Li Bin-Zhong
Huang Yan-Nv
Zeng Rong
Jing Nai-He
Chen Taiping
Li En
Sasaki Hiroyuki
Xu Guo-Liang
References (50)
50 references, click to expand
  1. The fundamental role of epigenetic events in cancer.
    Nat Rev Genet. 2002 Jun;3(6):415-28 PMID: 12042769
  2. Chromatin modification and epigenetic reprogramming in mammalian development.
    Nat Rev Genet. 2002 Sep;3(9):662-73 PMID: 12209141
  3. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors.
    Cell. 2006 Aug 25;126(4):663-76 PMID: 16904174
  4. Structure of Dnmt3a bound to Dnmt3L suggests a model for de novo DNA methylation.
    Nature. 2007 Sep 13;449(7159):248-51 PMID: 17713477
  5. Transposon silencing and imprint establishment in mammalian germ cells.
    Cold Spring Harb Symp Quant Biol. 2004;69:381-7 PMID: 16117671
  6. DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development.
    Cell. 1999 Oct 29;99(3):247-57 PMID: 10555141
  7. Dnmt3a and Dnmt3b are transcriptional repressors that exhibit unique localization properties to heterochromatin.
    J Biol Chem. 2001 Aug 24;276(34):32282-7 PMID: 11427539
  8. Roles for Dnmt3b in mammalian development: a mouse model for the ICF syndrome.
    Development. 2006 Mar;133(6):1183-92 PMID: 16501171
  9. Biotin-avidin microplate assay for the quantitative analysis of enzymatic methylation of DNA by DNA methyltransferases.
    Biol Chem. 2000 Mar;381(3):269-72 PMID: 10782999
  10. Maintenance and regulation of DNA methylation patterns in mammals.
    Biochem Cell Biol. 2005 Aug;83(4):438-48 PMID: 16094447
  11. Stage- and cell-specific expression of Dnmt3a and Dnmt3b during embryogenesis.
    Mech Dev. 2002 Oct;118(1-2):187-90 PMID: 12351185
  12. Physical and functional interactions between the human DNMT3L protein and members of the de novo methyltransferase family.
    J Cell Biochem. 2005 Aug 1;95(5):902-17 PMID: 15861382
  13. Oct4 is required for primordial germ cell survival.
    EMBO Rep. 2004 Nov;5(11):1078-83 PMID: 15486564
  14. De novo DNA methyltransferases Dnmt3a and Dnmt3b primarily mediate the cytotoxic effect of 5-aza-2'-deoxycytidine.
    Oncogene. 2005 Apr 28;24(19):3091-9 PMID: 15735669
  15. Targeted mutation of the DNA methyltransferase gene results in embryonic lethality.
    Cell. 1992 Jun 12;69(6):915-26 PMID: 1606615
  16. The DNA methyltransferases associate with HP1 and the SUV39H1 histone methyltransferase.
    Nucleic Acids Res. 2003 May 1;31(9):2305-12 PMID: 12711675
  17. Incomplete reactivation of Oct4-related genes in mouse embryos cloned from somatic nuclei.
    Development. 2003 Apr;130(8):1673-80 PMID: 12620990
  18. DNMT3L connects unmethylated lysine 4 of histone H3 to de novo methylation of DNA.
    Nature. 2007 Aug 9;448(7154):714-7 PMID: 17687327
  19. A novel Dnmt3a isoform produced from an alternative promoter localizes to euchromatin and its expression correlates with active de novo methylation.
    J Biol Chem. 2002 Oct 11;277(41):38746-54 PMID: 12138111
  20. Dnmt3L cooperates with the Dnmt3 family of de novo DNA methyltransferases to establish maternal imprints in mice.
    Development. 2002 Apr;129(8):1983-93 PMID: 11934864
  21. Essential role for de novo DNA methyltransferase Dnmt3a in paternal and maternal imprinting.
    Nature. 2004 Jun 24;429(6994):900-3 PMID: 15215868
  22. DNA demethylation is necessary for the epigenetic reprogramming of somatic cell nuclei.
    Nat Cell Biol. 2004 Oct;6(10):984-90 PMID: 15448701
  23. Cloning and sequencing of a cDNA encoding DNA methyltransferase of mouse cells. The carboxyl-terminal domain of the mammalian enzymes is related to bacterial restriction methyltransferases.
    J Mol Biol. 1988 Oct 20;203(4):971-83 PMID: 3210246
  24. Mutations in DNA methyltransferase DNMT3B in ICF syndrome affect its regulation by DNMT3L.
    Hum Mol Genet. 2006 May 1;15(9):1375-85 PMID: 16543361
  25. Mouse germline restriction of Oct4 expression by germ cell nuclear factor.
    Dev Cell. 2001 Sep;1(3):377-87 PMID: 11702949
  26. Chromosome instability and immunodeficiency syndrome caused by mutations in a DNA methyltransferase gene.
    Nature. 1999 Nov 11;402(6758):187-91 PMID: 10647011
  27. A modified and improved method for bisulphite based cytosine methylation analysis.
    Nucleic Acids Res. 1996 Dec 15;24(24):5064-6 PMID: 9016686
  28. Oct4 distribution and level in mouse clones: consequences for pluripotency.
    Genes Dev. 2002 May 15;16(10):1209-19 PMID: 12023300
  29. Imprinting regulator DNMT3L is a transcriptional repressor associated with histone deacetylase activity.
    Nucleic Acids Res. 2002 Aug 15;30(16):3602-8 PMID: 12177302
  30. Co-operation and communication between the human maintenance and de novo DNA (cytosine-5) methyltransferases.
    EMBO J. 2002 Aug 1;21(15):4183-95 PMID: 12145218
  31. The PWWP domain of Dnmt3a and Dnmt3b is required for directing DNA methylation to the major satellite repeats at pericentric heterochromatin.
    Mol Cell Biol. 2004 Oct;24(20):9048-58 PMID: 15456878
  32. A targeting sequence directs DNA methyltransferase to sites of DNA replication in mammalian nuclei.
    Cell. 1992 Nov 27;71(5):865-73 PMID: 1423634
  33. Core filaments of the nuclear matrix.
    J Cell Biol. 1990 Mar;110(3):569-80 PMID: 2307700
  34. G9a-mediated irreversible epigenetic inactivation of Oct-3/4 during early embryogenesis.
    Nat Cell Biol. 2006 Feb;8(2):188-94 PMID: 16415856
  35. Human DNA-(cytosine-5) methyltransferase-PCNA complex as a target for p21WAF1.
    Science. 1997 Sep 26;277(5334):1996-2000 PMID: 9302295
  36. Mechanism of stimulation of catalytic activity of Dnmt3A and Dnmt3B DNA-(cytosine-C5)-methyltransferases by Dnmt3L.
    J Biol Chem. 2005 Apr 8;280(14):13341-8 PMID: 15671018
  37. Extinction of Oct-3/4 gene expression in embryonal carcinoma x fibroblast somatic cell hybrids is accompanied by changes in the methylation status, chromatin structure, and transcriptional activity of the Oct-3/4 upstream region.
    Mol Cell Biol. 1993 Feb;13(2):891-901 PMID: 7678695
  38. Imaging molecular interactions in living cells.
    Mol Endocrinol. 2005 Jul;19(7):1675-86 PMID: 15761028
  39. The DNA methyltransferases of mammals.
    Hum Mol Genet. 2000 Oct;9(16):2395-402 PMID: 11005794
  40. The DNA methyltransferase-like protein DNMT3L stimulates de novo methylation by Dnmt3a.
    Proc Natl Acad Sci U S A. 2002 Dec 24;99(26):16916-21 PMID: 12481029
  41. Six3, a medaka homologue of the Drosophila homeobox gene sine oculis is expressed in the anterior embryonic shield and the developing eye.
    Mech Dev. 1998 Jun;74(1-2):159-64 PMID: 9651515
  42. Dnmt3L and the establishment of maternal genomic imprints.
    Science. 2001 Dec 21;294(5551):2536-9 PMID: 11719692
  43. Differential recruitment of methylated CpG binding domains by the orphan receptor GCNF initiates the repression and silencing of Oct4 expression.
    Mol Cell Biol. 2006 Dec;26(24):9471-83 PMID: 17030610
  44. De novo DNA cytosine methyltransferase activities in mouse embryonic stem cells.
    Development. 1996 Oct;122(10):3195-205 PMID: 8898232
  45. Establishment and maintenance of genomic methylation patterns in mouse embryonic stem cells by Dnmt3a and Dnmt3b.
    Mol Cell Biol. 2003 Aug;23(16):5594-605 PMID: 12897133
  46. Chromatin targeting of de novo DNA methyltransferases by the PWWP domain.
    J Biol Chem. 2004 Jun 11;279(24):25447-54 PMID: 14998998
  47. Cloning and characterization of a family of novel mammalian DNA (cytosine-5) methyltransferases.
    Nat Genet. 1998 Jul;19(3):219-20 PMID: 9662389
  48. DNA methylation patterns and epigenetic memory.
    Genes Dev. 2002 Jan 1;16(1):6-21 PMID: 11782440
  49. A generic protein purification method for protein complex characterization and proteome exploration.
    Nat Biotechnol. 1999 Oct;17(10):1030-2 PMID: 10504710
  50. Dnmt3a binds deacetylases and is recruited by a sequence-specific repressor to silence transcription.
    EMBO J. 2001 May 15;20(10):2536-44 PMID: 11350943
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
1098-5549
DOI
10.1128/MCB.01380-07
Published
2007-12-00
Epub
2007-00-15
Pages
8748-59
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC2169413
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.01380-07 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2