Molecular Biology Reports

, Volume 40, Issue 2, pp 797–801 | Cite as

The methylation of C/EBP β gene promoter and regulated by GATA-2 protein

Article

Abstract

Mammalian genomes are punctuated by DNA sequences containing an atypically high frequency of CpG sites (CpG islands; CGIs) that are associated with the majority of annotated gene promoters. Methylated C bases of CpG sites inhibit the expression of downstream genes. During the differentiation of 3T3-L1 preadipocytes, the CCAAT/enhancer-binding protein (C/EBP) β gene plays an important role. We studied the CpG island methylation status of the C/EBP β promoter and its relationship with the GATA-2 protein. We used computer analysis to determine that the C/EBP β promoter sequence is rich in CGIs, and observed that two of seven methylated C bases were demethylated during the preadipocyte differentiation using bisulfite sequencing PCR (BSP). This corresponded with the onset of notable C/EBP β gene expression. Immunofluorescence and molecular docking showed that the GATA-2 protein binds the C/EBP β promoter in front of the first demethylated CpG site. We also found that expression of GATA-2 and C/EBP β proteins is negatively correlated. These results indicate that the methylated C bases in the C/EBP β promoter relate to expression of the C/EBP β gene, and that its demethylation is linked with GATA-2 protein association.

Keywords

CpG island GATA-2 C/EBP β Preadipocyte Methylation 

References

  1. 1.
    Illingworth RS, Bird AP (2009) CpG islands—‘a rough guide’. FEBS Lett 583:1713–1720PubMedCrossRefGoogle Scholar
  2. 2.
    Pfeifer GP (2006) Mutagenesis at methylated CpG sequences. Curr Top Microbiol Immunol 301:259–281PubMedCrossRefGoogle Scholar
  3. 3.
    Zhang N, Lin C, Huang X, Kolbanovskiy A, Hingerty BE, Amin S, Broyde S, Geacintov NE, Patel DJ (2005) Methylation of cytosine at C5 in a CpG sequence context causes a conformational switch of a benzo-pyrene diolepoxide-N2-guanine adduct in DNA from a minor groove alignment to intercalation with base displacement. J Mol Biol 346:951–965PubMedCrossRefGoogle Scholar
  4. 4.
    Tang QQ, Lane MD (1999) Activation and centromeric localization of CCAAT/enhancer-binding proteins during the mitotic clonal expansion of adipocyte differentiation. Genes Dev 13:2231–2241PubMedCrossRefGoogle Scholar
  5. 5.
    Lane MD, Tang QQ, Jiang MS (1999) Role of the CCAAT enhancer binding proteins (C/EBPs) in adipocyte differentiation. Biochem Biophys Res Commun 266:677–683PubMedCrossRefGoogle Scholar
  6. 6.
    Yeh WC, Cao Z, Mcknight SL (1995) Cascade regulation of terminal adipocyte differentiation by three members of the C/EBP family of leucine zipper proteins. Genes Dev 15:168–181CrossRefGoogle Scholar
  7. 7.
    Tanaka T, Yoshida N, Kishimoto T, Akira (1997) Defective adipocyte differentiation in mice lacking the C/EBP beta and/or C/EBP delta gene. EMBO J 16:7432–7443PubMedCrossRefGoogle Scholar
  8. 8.
    Kornhauser JM, Leonard MW, Yamamoto M, LaVail JH, Mayo KE, Engel JD (1994) Temporal and spatial changes in GATA transcription factor expression are coincident with development of the chicken optic tectum. Brain Res Mol Brain Res 23:100–110PubMedCrossRefGoogle Scholar
  9. 9.
    Tong Q, Tsai J, Tan G, Dalgin G, Hotamisligil GS (2005) Interaction between GATA and the C/EBP family of transcription factors is critical in GATA-mediated suppression of adipocyte differentiation. Mol Cell Biol 25:706–715PubMedCrossRefGoogle Scholar
  10. 10.
    Tong Q, Dalgin G, Xu H, Ting CN, Leiden JM, Hotamisligil GS (2000) Function of GATA transcription factors in preadipocyte-adipocyte transition. Science 5489:134–138CrossRefGoogle Scholar
  11. 11.
    Linnemann AK, O’Geen H, Keles S, Farnham PJ, Bresnick EH (2011) Genetic framework for GATA factor function in vascular biology. Proc Natl Acad Sci USA 108:13641–13646PubMedCrossRefGoogle Scholar
  12. 12.
    Zheng R, Blobel GA (2010) GATA transcription factors and cancer. Genes Cancer 1:1178–1188PubMedCrossRefGoogle Scholar
  13. 13.
    Pedone PV, Omichinski JG, Nony P, Trainor C, Gronenborn AM, Clore GM, Felsenfeld G (1997) The N-terminal fingers of chicken GATA-2 and GATA-3 are independent sequence-specific DNA binding domains. EMBO J 16:2874–2882PubMedCrossRefGoogle Scholar
  14. 14.
    Visvader JE, Crossley M, Hill J, Orkin SH, Adams JM (1995) The C-terminal zinc finger of GATA-1 or GATA-2 is sufficient to induce megakaryocytic differentiation of an early myeloid cell line. Mol Cell Biol 15:634–641PubMedGoogle Scholar
  15. 15.
    Choi BH, Ahn IS, Kim YH, Park JW, Lee SY, Hyun CK, Do MS (2006) Berberine reduces the expression of adipogenic enzymes and inflammatory molecules of 3T3-L1 adipocyte. Exp Mol Med 38:599–605PubMedGoogle Scholar
  16. 16.
    Huang C, Zhang YB, Gong ZW, Sheng XY, Li ZM, Zhang Qin Y (2006) Berberine inhibits 3T3-L1 adipocyte differentiation through the PPARr pathway. Biochem Biophys Res Commun 348:571–578PubMedCrossRefGoogle Scholar
  17. 17.
    KO LJ, Engel JD (1993) DNA-binding specificities of the GATA transcription factor family. Mol Cell Biol 13:4011–4022PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  1. 1.Department of Molecular GeneticsShanghai Medical School, Fudan University, Key Laboratory of Molecular Medicine, Ministry of EducationShanghaiChina

Personalised recommendations