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Nucleic Acids Res. Oct 25, 1991; 19(20): 5771–5775.
PMCID: PMC328989

Estradiol down regulates the binding activity of an avian vitellogenin gene repressor (MDBP-2) and triggers a gradual demethylation of the mCpG pair of its DNA binding site.


A negative regulating protein (MDBP-2) from rooster liver nuclear extracts binds preferentially to a methylated promoter region 5'TTCACCTTmCGCTATGAGGGGGATCATACTGG3' of the avian vitellogenin II gene (Nucleic Acids Res. 19, 1029-1034, 1991). Treatment of adult and immature roosters with estradiol results in a 90% decrease in the binding activity of MDBP-2 within three days. This corresponds to the level found in egg laying hens. The decrease in the binding activity of MDBP-2 precedes the onset of vitellogenin gene transcription. At the same time, there is a two-fold increase in the binding activity of NHP-1 (tested with the same oligonucleotide as for MDBP-2), a protein thought to be involved in the active demethylation of DNA. The methylated oligonucleotide binds either MDBP-2 or NHP-1 and there is no complex formation between the two proteins and DNA. Estradiol treatment does not change the equilibrium binding constant of MDBP-2 which is about 10(-9)M for the methylated oligonucleotide. The early kinetics of demethylation of the mCpG pair in the binding site of MDBP-2 was studied by means of genomic sequencing. A low level of demethylation of mCpG starts gradually on both DNA strands already 4 hours after estradiol treatment during the lag phase of vitellogenin mRNA synthesis. It is concluded that the lowering of the binding activity of MDBP-2 may have a stronger effect on the derepression of the gene than the slow demethylation of MDBP-2 DNA binding site. The role of the methylated CpG is to assure a high binding affinity of the repressor to DNA.

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Selected References

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  • Wilks AF, Cozens PJ, Mattaj IW, Jost JP. Estrogen induces a demethylation at the 5' end region of the chicken vitellogenin gene. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4252–4255. [PMC free article] [PubMed]
  • Meijlink FC, Philipsen JN, Gruber M, Ab G. Methylation of the chicken vitellogenin gene: influence of estradiol administration. Nucleic Acids Res. 1983 Mar 11;11(5):1361–1373. [PMC free article] [PubMed]
  • Saluz HP, Jiricny J, Jost JP. Genomic sequencing reveals a positive correlation between the kinetics of strand-specific DNA demethylation of the overlapping estradiol/glucocorticoid-receptor binding sites and the rate of avian vitellogenin mRNA synthesis. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7167–7171. [PMC free article] [PubMed]
  • Saluz HP, Feavers IM, Jiricny J, Jost JP. Genomic sequencing and in vivo footprinting of an expression-specific DNase I-hypersensitive site of avian vitellogenin II promoter reveal a demethylation of a mCpG and a change in specific interactions of proteins with DNA. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6697–6700. [PMC free article] [PubMed]
  • Jost JP, Seldran M. Association of transcriptionally active vitellogenin II gene with the nuclear matrix of chicken liver. EMBO J. 1984 Sep;3(9):2005–2008. [PMC free article] [PubMed]
  • Burch JB, Weintraub H. Temporal order of chromatin structural changes associated with activation of the major chicken vitellogenin gene. Cell. 1983 May;33(1):65–76. [PubMed]
  • Burch JB. Identification and sequence analysis of the 5' end of the major chicken vitellogenin gene. Nucleic Acids Res. 1984 Jan 25;12(2):1117–1135. [PMC free article] [PubMed]
  • Vaccaro M, Pawlak A, Jost JP. Positive and negative regulatory elements of chicken vitellogenin II gene characterized by in vitro transcription competition assays in a homologous system. Proc Natl Acad Sci U S A. 1990 Apr;87(8):3047–3051. [PMC free article] [PubMed]
  • Pawlak A, Bryans M, Jost JP. An avian 40 KDa nucleoprotein binds preferentially to a promoter sequence containing one single pair of methylated CpG. Nucleic Acids Res. 1991 Mar 11;19(5):1029–1034. [PMC free article] [PubMed]
  • Gorski K, Carneiro M, Schibler U. Tissue-specific in vitro transcription from the mouse albumin promoter. Cell. 1986 Dec 5;47(5):767–776. [PubMed]
  • Saluz H, Jost JP. A simple high-resolution procedure to study DNA methylation and in vivo DNA-protein interactions on a single-copy gene level in higher eukaryotes. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2602–2606. [PMC free article] [PubMed]
  • Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. [PubMed]
  • Thomas PS. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. [PMC free article] [PubMed]
  • Cozens PJ, Cato AC, Jost JP. Characterization of cloned complementary DNA covering more than 6000 nucleotides (97%) of avian vitellogenin mRNA. Eur J Biochem. 1980 Dec;112(3):443–450. [PubMed]
  • Hughes MJ, Jost JP. The ubiquitous nuclear protein, NHP1, binds with high affinity to different sequences of the chicken vitellogenin II gene. Nucleic Acids Res. 1989 Nov 11;17(21):8511–8520. [PMC free article] [PubMed]
  • Jost JP, Munch O, Andersson T. Study of protein-DNA interactions by surface plasmon resonance (real time kinetics). Nucleic Acids Res. 1991 May 25;19(10):2788–2788. [PMC free article] [PubMed]
  • Beuving G, Gruber M. Induction of phosvitin synthesis in roosters by estradiol injection. Biochim Biophys Acta. 1971 Mar 25;232(3):529–536. [PubMed]
  • Deeley RG, Udell DS, Burns AT, Gordon JI, Goldberger RF. Kinetics of avian vitellogenin messenger RNA induction. Comparison between primary and secondary response to estrogen. J Biol Chem. 1977 Nov 25;252(22):7913–7915. [PubMed]
  • Jost JP, Ohno T, Panyim S, Schuerch AR. Appearance of vitellogenin mRNA sequences and rate of vitellogenin synthesis in chicken liver following primary and secondary stimulation by 17 beta-estradiol. Eur J Biochem. 1978 Mar 15;84(2):355–361. [PubMed]
  • Schröcksnadel H, Bator A, Frick J. Plasma estradiol-17beta level in the domestic fowl. Steroids. 1973 Dec;22(6):767–773. [PubMed]
  • Senior BE. Oestradiol concentration in the peripheral plasma of the domestic hen from 7 weeks of age until the time of sexual maturity. J Reprod Fertil. 1974 Nov;41(1):107–112. [PubMed]
  • Joss U, Bassand C, Dierks-Ventling C. Rapid appearance of estrogen receptor in chick liver nuclei: partial inhibition by cycloheximide. FEBS Lett. 1976 Jul 15;66(2):293–298. [PubMed]
  • McEwan IJ, Saluz HP, Jost JP. In vivo and in vitro protein-DNA interactions at the distal oestrogen response element of the chicken vitellogenin gene: evidence for the same protein binding to this sequence in hen and rooster liver. J Steroid Biochem Mol Biol. 1991 Mar;38(3):275–283. [PubMed]
  • Boyes J, Bird A. DNA methylation inhibits transcription indirectly via a methyl-CpG binding protein. Cell. 1991 Mar 22;64(6):1123–1134. [PubMed]
  • Levine A, Cantoni GL, Razin A. Inhibition of promoter activity by methylation: possible involvement of protein mediators. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6515–6518. [PMC free article] [PubMed]
  • Corthésy B, Cardinaux JR, Claret FX, Wahli W. A nuclear factor I-like activity and a liver-specific repressor govern estrogen-regulated in vitro transcription from the Xenopus laevis vitellogenin B1 promoter. Mol Cell Biol. 1989 Dec;9(12):5548–5562. [PMC free article] [PubMed]
  • Seal SN, Davis DL, Burch JB. Mutational studies reveal a complex set of positive and negative control elements within the chicken vitellogenin II promoter. Mol Cell Biol. 1991 May;11(5):2704–2717. [PMC free article] [PubMed]
  • Hughes MJ, Liang HM, Jiricny J, Jost JP. Purification and characterization of a protein from HeLa cells that binds with high affinity to the estrogen response element, GGTCAGCGTGACC. Biochemistry. 1989 Nov 14;28(23):9137–9142. [PubMed]
  • Razin A, Szyf M, Kafri T, Roll M, Giloh H, Scarpa S, Carotti D, Cantoni GL. Replacement of 5-methylcytosine by cytosine: a possible mechanism for transient DNA demethylation during differentiation. Proc Natl Acad Sci U S A. 1986 May;83(9):2827–2831. [PMC free article] [PubMed]

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