Home LiteratureArticle Details
PMID: 15923607 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Reduced genomic cytosine methylation and defective cellular differentiation in embryonic stem cells lacking CpG binding protein.

Molecular and cellular biology ·Vol. 25 ·No. 12 ·2005-06-00 ·Pages 4881-91

Carlone DL, Lee JH, Young SR, Dobrota E, Butler JS, Ruiz J, Skalnik DG

Abstract

Cytosine methylation at CpG dinucleotides is a critical epigenetic modification of mammalian genomes. CpG binding protein (CGBP) exhibits a unique DNA-binding specificity for unmethylated CpG motifs and is essential for early murine development. Embryonic stem cell lines deficient for CGBP were generated to further examine CGBP function. CGBP(-)(/)(-) cells are viable but show an increased rate of apoptosis and are unable to achieve in vitro differentiation following removal of leukemia inhibitory factor from the growth media. Instead, CGBP(-)(/)(-) embryonic stem cells remain undifferentiated as revealed by persistent expression of the pluripotent markers Oct4 and alkaline phosphatase. CGBP(-)(/)(-) cells exhibit a 60 to 80% decrease in global cytosine methylation, including hypo-methylation of repetitive elements, single-copy genes, and imprinted genes. Total DNA methyltransferase activity is reduced by 30 to 60% in CGBP(-)(/)(-) cells, and expression of the maintenance DNA methyltransferase 1 protein is similarly reduced. However, de novo DNA methyltransferase activity is normal. Nearly all aspects of the pleiotropic CGBP(-)(/)(-) phenotype are rescued by introduction of a CGBP expression vector. Hence, CGBP is essential for normal epigenetic modification of the genome by cytosine methylation and for cellular differentiation, consistent with the requirement for CGBP during early mammalian development.

MeSH Terms
Animals Apoptosis Cell Differentiation/physiology CpG Islands Cytosine/metabolism DNA (Cytosine-5-)-Methyltransferase 1 DNA (Cytosine-5-)-Methyltransferases/metabolism DNA Methylation DNA-Binding Proteins/genetics,metabolism Epigenesis, Genetic Female Fetal Viability Humans Male Mice Mice, Knockout Phenotype Pregnancy Stem Cells/physiology Trans-Activators/genetics,metabolism
Chemicals
Cxxc1 protein, mouse DNA-Binding Proteins Trans-Activators Cytosine DNA (Cytosine-5-)-Methyltransferase 1 DNA (Cytosine-5-)-Methyltransferases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Carlone Diana L
Herman B Wells Center for Pediatric Research, Section of Pediatric Hematology/Oncology, Department of Pediatrics, Indiana School of Medicine, Indianapolis, IN 46202, USA.
Lee Jeong-Heon
Young Suzanne R L
Dobrota Erika
Butler Jill Sergesketter
Ruiz Joseph
Skalnik David G
References (80)
80 references, click to expand
  1. Chromatin modification and epigenetic reprogramming in mammalian development.
    Nat Rev Genet. 2002 Sep;3(9):662-73 PMID: 12209141
  2. A unique developmental pattern of Oct-3/4 DNA methylation is controlled by a cis-demodification element.
    J Biol Chem. 2002 Sep 13;277(37):34521-30 PMID: 12110668
  3. Cloning and characterization of the gene encoding the mouse homologue of CpG binding protein.
    Gene. 2002 Jul 24;295(1):71-7 PMID: 12242013
  4. CpG-binding protein is a nuclear matrix- and euchromatin-associated protein localized to nuclear speckles containing human trithorax. Identification of nuclear matrix targeting signals.
    J Biol Chem. 2002 Nov 1;277(44):42259-67 PMID: 12200428
  5. ALL-1 is a histone methyltransferase that assembles a supercomplex of proteins involved in transcriptional regulation.
    Mol Cell. 2002 Nov;10(5):1119-28 PMID: 12453419
  6. High conservation of the Set1/Rad6 axis of histone 3 lysine 4 methylation in budding and fission yeasts.
    J Biol Chem. 2003 Mar 7;278(10):8487-93 PMID: 12488447
  7. Human Sin3 deacetylase and trithorax-related Set1/Ash2 histone H3-K4 methyltransferase are tethered together selectively by the cell-proliferation factor HCF-1.
    Genes Dev. 2003 Apr 1;17(7):896-911 PMID: 12670868
  8. Epigenomic stress response. Knockdown of DNA methyltransferase 1 triggers an intra-S-phase arrest of DNA replication and induction of stress response genes.
    J Biol Chem. 2003 Apr 25;278(17):14985-95 PMID: 12576480
  9. The DNA methyltransferases associate with HP1 and the SUV39H1 histone methyltransferase.
    Nucleic Acids Res. 2003 May 1;31(9):2305-12 PMID: 12711675
  10. Trimethylated lysine 9 of histone H3 is a mark for DNA methylation in Neurospora crassa.
    Nat Genet. 2003 May;34(1):75-9 PMID: 12679815
  11. The PHD finger of the chromatin-associated protein ING2 functions as a nuclear phosphoinositide receptor.
    Cell. 2003 Jul 11;114(1):99-111 PMID: 12859901
  12. Suv39h-mediated histone H3 lysine 9 methylation directs DNA methylation to major satellite repeats at pericentric heterochromatin.
    Curr Biol. 2003 Jul 15;13(14):1192-200 PMID: 12867029
  13. 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
  14. Histone deacetylase activity is required for embryonic stem cell differentiation.
    Genesis. 2004 Jan;38(1):32-8 PMID: 14755802
  15. Epigenetic control of mouse Oct-4 gene expression in embryonic stem cells and trophoblast stem cells.
    J Biol Chem. 2004 Apr 23;279(17):17063-9 PMID: 14761969
  16. Leukemia proto-oncoprotein MLL forms a SET1-like histone methyltransferase complex with menin to regulate Hox gene expression.
    Mol Cell Biol. 2004 Jul;24(13):5639-49 PMID: 15199122
  17. Human DNA methyltransferase 1 is required for maintenance of the histone H3 modification pattern.
    J Biol Chem. 2004 Aug 27;279(35):37175-84 PMID: 15220328
  18. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
    Nucleic Acids Res. 1983 Mar 11;11(5):1475-89 PMID: 6828386
  19. Temporal and regional changes in DNA methylation in the embryonic, extraembryonic and germ cell lineages during mouse embryo development.
    Development. 1987 Mar;99(3):371-82 PMID: 3653008
  20. Activation of mammalian DNA methyltransferase by cleavage of a Zn binding regulatory domain.
    EMBO J. 1992 Jul;11(7):2611-7 PMID: 1628623
  21. Involvement of a homolog of Drosophila trithorax by 11q23 chromosomal translocations in acute leukemias.
    Cell. 1992 Nov 13;71(4):691-700 PMID: 1423624
  22. The t(4;11) chromosome translocation of human acute leukemias fuses the ALL-1 gene, related to Drosophila trithorax, to the AF-4 gene.
    Cell. 1992 Nov 13;71(4):701-8 PMID: 1423625
  23. Hematopoietic commitment during embryonic stem cell differentiation in culture.
    Mol Cell Biol. 1993 Jan;13(1):473-86 PMID: 8417345
  24. Maternal-specific methylation of the imprinted mouse Igf2r locus identifies the expressed locus as carrying the imprinting signal.
    Cell. 1993 Apr 9;73(1):61-71 PMID: 8462104
  25. Hematopoietic growth factor receptor genes as markers of lineage commitment during in vitro development of hematopoietic cells.
    Blood. 1993 Jun 1;81(11):2903-15 PMID: 8499630
  26. DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development.
    Cell. 1999 Oct 29;99(3):247-57 PMID: 10555141
  27. DNA methyltransferase Dnmt1 associates with histone deacetylase activity.
    Nat Genet. 2000 Jan;24(1):88-91 PMID: 10615135
  28. Chromosome instability and immunodeficiency syndrome caused by mutations in a DNA methyltransferase gene.
    Nature. 1999 Nov 11;402(6758):187-91 PMID: 10647011
  29. Cloning of a mammalian transcriptional activator that binds unmethylated CpG motifs and shares a CXXC domain with DNA methyltransferase, human trithorax, and methyl-CpG binding domain protein 1.
    Mol Cell Biol. 2000 Mar;20(6):2108-21 PMID: 10688657
  30. Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells.
    Nat Genet. 2000 Apr;24(4):372-6 PMID: 10742100
  31. DNMT1 forms a complex with Rb, E2F1 and HDAC1 and represses transcription from E2F-responsive promoters.
    Nat Genet. 2000 Jul;25(3):338-42 PMID: 10888886
  32. Loss of genomic methylation causes p53-dependent apoptosis and epigenetic deregulation.
    Nat Genet. 2001 Jan;27(1):31-9 PMID: 11137995
  33. Loss of the maintenance methyltransferase, xDnmt1, induces apoptosis in Xenopus embryos.
    EMBO J. 2001 Apr 17;20(8):1963-73 PMID: 11296229
  34. 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
  35. Epigenetic reprogramming in mammalian development.
    Science. 2001 Aug 10;293(5532):1089-93 PMID: 11498579
  36. Reduced rates of gene loss, gene silencing, and gene mutation in Dnmt1-deficient embryonic stem cells.
    Mol Cell Biol. 2001 Nov;21(22):7587-600 PMID: 11604495
  37. CpG binding protein is crucial for early embryonic development.
    Mol Cell Biol. 2001 Nov;21(22):7601-6 PMID: 11604496
  38. COMPASS: a complex of proteins associated with a trithorax-related SET domain protein.
    Proc Natl Acad Sci U S A. 2001 Nov 6;98(23):12902-7 PMID: 11687631
  39. Polycomblike PHD fingers mediate conserved interaction with enhancer of zeste protein.
    J Biol Chem. 2001 Nov 16;276(46):43065-73 PMID: 11571280
  40. Lsh, a member of the SNF2 family, is required for genome-wide methylation.
    Genes Dev. 2001 Nov 15;15(22):2940-4 PMID: 11711429
  41. Identification and characterization of the DNA binding domain of CpG-binding protein.
    J Biol Chem. 2001 Nov 30;276(48):44669-76 PMID: 11572867
  42. Dnmt1 overexpression causes genomic hypermethylation, loss of imprinting, and embryonic lethality.
    Mol Cell Biol. 2002 Apr;22(7):2124-35 PMID: 11884600
  43. DNA methyltransferases get connected to chromatin.
    Trends Genet. 2002 Jun;18(6):275-7 PMID: 12044346
  44. LCX, leukemia-associated protein with a CXXC domain, is fused to MLL in acute myeloid leukemia with trilineage dysplasia having t(10;11)(q22;q23).
    Cancer Res. 2002 Jul 15;62(14):4075-80 PMID: 12124344
  45. DNA methylation in folate deficiency: use of CpG methylase.
    Biochem Biophys Res Commun. 1993 Jun 30;193(3):1184-90 PMID: 8323540
  46. Analysis of the murine All-1 gene reveals conserved domains with human ALL-1 and identifies a motif shared with DNA methyltransferases.
    Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6350-4 PMID: 8327517
  47. Acute mixed-lineage leukemia t(4;11)(q21;q23) generates an MLL-AF4 fusion product.
    Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7884-8 PMID: 7689231
  48. In vitro chamber specification during embryonic stem cell cardiogenesis. Expression of the ventricular myosin light chain-2 gene is independent of heart tube formation.
    J Biol Chem. 1993 Nov 25;268(33):25244-52 PMID: 8227090
  49. Developmental changes in methylation of spermatogenesis-specific genes include reprogramming in the epididymis.
    Nat Genet. 1994 May;7(1):59-63 PMID: 8075642
  50. DNA methyltransferases.
    Curr Opin Cell Biol. 1994 Jun;6(3):380-9 PMID: 7917329
  51. 11q23 translocations split the "AT-hook" cruciform DNA-binding region and the transcriptional repression domain from the activation domain of the mixed-lineage leukemia (MLL) gene.
    Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10610-4 PMID: 7938000
  52. The PHD finger: implications for chromatin-mediated transcriptional regulation.
    Trends Biochem Sci. 1995 Feb;20(2):56-9 PMID: 7701562
  53. Domains with transcriptional regulatory activity within the ALL1 and AF4 proteins involved in acute leukemia.
    Proc Natl Acad Sci U S A. 1995 Dec 19;92(26):12160-4 PMID: 8618864
  54. Germ-line passage is required for establishment of methylation and expression patterns of imprinted but not of nonimprinted genes.
    Genes Dev. 1996 Apr 15;10(8):1008-20 PMID: 8608936
  55. De novo DNA cytosine methyltransferase activities in mouse embryonic stem cells.
    Development. 1996 Oct;122(10):3195-205 PMID: 8898232
  56. Complementation of methylation deficiency in embryonic stem cells by a DNA methyltransferase minigene.
    Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):12920-5 PMID: 8917520
  57. A component of the transcriptional repressor MeCP1 shares a motif with DNA methyltransferase and HRX proteins.
    Nat Genet. 1997 Jul;16(3):256-9 PMID: 9207790
  58. Retinoic acid accelerates embryonic stem cell-derived cardiac differentiation and enhances development of ventricular cardiomyocytes.
    J Mol Cell Cardiol. 1997 Jun;29(6):1525-39 PMID: 9220339
  59. A 5' 2-kilobase-pair region of the imprinted mouse H19 gene exhibits exclusive paternal methylation throughout development.
    Mol Cell Biol. 1997 Aug;17(8):4322-9 PMID: 9234689
  60. Cytosine methylation and the ecology of intragenomic parasites.
    Trends Genet. 1997 Aug;13(8):335-40 PMID: 9260521
  61. Constitutive and inducible expression of megakaryocyte-specific genes in Friend erythroleukaemia cells.
    Br J Haematol. 1997 Dec;99(3):500-8 PMID: 9401056
  62. Embryonic germ cells induce epigenetic reprogramming of somatic nucleus in hybrid cells.
    EMBO J. 1997 Nov 3;16(21):6510-20 PMID: 9351832
  63. Specific arrest of cardiogenesis in cultured embryonic stem cells lacking Cripto-1.
    Dev Biol. 1998 Apr 15;196(2):237-47 PMID: 9576836
  64. The DNA methylation locus DDM1 is required for maintenance of gene silencing in Arabidopsis.
    Genes Dev. 1998 Jun 1;12(11):1714-25 PMID: 9620857
  65. Trichostatin A causes selective loss of DNA methylation in Neurospora.
    Proc Natl Acad Sci U S A. 1998 Aug 4;95(16):9430-5 PMID: 9689097
  66. Transcription of IAP endogenous retroviruses is constrained by cytosine methylation.
    Nat Genet. 1998 Oct;20(2):116-7 PMID: 9771701
  67. Identification and characterization of a family of mammalian methyl-CpG binding proteins.
    Mol Cell Biol. 1998 Nov;18(11):6538-47 PMID: 9774669
  68. Deletion of the H19 differentially methylated domain results in loss of imprinted expression of H19 and Igf2.
    Genes Dev. 1998 Dec 1;12(23):3693-702 PMID: 9851976
  69. The sins of the fathers and mothers: genomic imprinting in mammalian development.
    Cell. 1999 Jan 22;96(2):185-93 PMID: 9988214
  70. Identification and functional characterization of the murine Rac2 gene promoter.
    DNA Cell Biol. 1999 Mar;18(3):253-63 PMID: 10098607
  71. Maintenance of genomic methylation requires a SWI2/SNF2-like protein.
    Nat Genet. 1999 May;22(1):94-7 PMID: 10319870
  72. DNA methylation.
    Blood. 1999 Jun 15;93(12):4059-70 PMID: 10361102
  73. MLL2: A new mammalian member of the trx/MLL family of genes.
    Genomics. 1999 Jul 15;59(2):187-92 PMID: 10409430
  74. Analysis of the NuRD subunits reveals a histone deacetylase core complex and a connection with DNA methylation.
    Genes Dev. 1999 Aug 1;13(15):1924-35 PMID: 10444591
  75. MBD2 is a transcriptional repressor belonging to the MeCP1 histone deacetylase complex.
    Nat Genet. 1999 Sep;23(1):58-61 PMID: 10471499
  76. Rett syndrome is caused by mutations in X-linked MECP2, encoding methyl-CpG-binding protein 2.
    Nat Genet. 1999 Oct;23(2):185-8 PMID: 10508514
  77. Expression pattern of the mouse T gene and its role in mesoderm formation.
    Nature. 1990 Feb 15;343(6259):657-9 PMID: 1689462
  78. In vivo footprint and methylation analysis by PCR-aided genomic sequencing: comparison of active and inactive X chromosomal DNA at the CpG island and promoter of human PGK-1.
    Genes Dev. 1990 Aug;4(8):1277-87 PMID: 2227409
  79. Growth regulation of mouse DNA methyltransferase gene expression.
    J Biol Chem. 1991 Jun 5;266(16):10027-30 PMID: 2037559
  80. Targeted mutation of the DNA methyltransferase gene results in embryonic lethality.
    Cell. 1992 Jun 12;69(6):915-26 PMID: 1606615
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.25.12.4881-4891.2005
Published
2005-06-00
Pages
4881-91
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC1140577
Subset
IM
Grants
NHLBI NIH HHS · P01 HL069974 · United States
NHLBI NIH HHS · HL69974 · United States
External Links
PubMed source DOI: 10.1128/MCB.25.12.4881-4891.2005 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: [email protected]

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