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PMID: 1695733 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

CpG methylation inhibits proenkephalin gene expression and binding of the transcription factor AP-2.

Nucleic acids research ·Vol. 18 ·No. 13 ·1990-07-11 ·Pages 3975-82

Comb M, Goodman HM

Abstract

DNA methylation at HpaII (CmCGG) sites inhibits expression of a human proenkephalin-CAT fusion gene when it is transiently expressed in CV-1 cells or stably expressed in C6-glioma cells. The inhibitory effects of HpaII methylation have been mapped to a site within the human proenkephalin promoter located at position -72 relative to the start site of transcription. This region spans a cAMP and phorbol ester inducible enhancer and methylation at this position inhibits both basal transcription and transcription induced by either cAMP or TPA. The HpaII site is located within an element which binds the transcription factor AP-2. In vitro methylation at this HpaII site inhibits the binding of AP-2. These results suggest that CpG methylation inhibits proenkephalin gene expression by directly interfering with the binding of a positively acting transcription factor previously shown to be essential for maximal basal, cAMP, and TPA inducible transcription.

MeSH Terms
5-Methylcytosine Base Sequence Cloning, Molecular Cytosine/analogs & derivatives,metabolism DNA/metabolism DNA-Binding Proteins/genetics,metabolism Deoxyribonuclease HpaII Deoxyribonucleases, Type II Site-Specific/metabolism Dinucleoside Phosphates/metabolism Enkephalins/genetics Gene Expression Regulation Humans Methylation Molecular Sequence Data Promoter Regions, Genetic Protein Precursors/genetics Restriction Mapping Transcription Factor AP-2 Transcription Factors/genetics,metabolism Transcription, Genetic Tumor Cells, Cultured
Chemicals
DNA-Binding Proteins Dinucleoside Phosphates Enkephalins Protein Precursors Transcription Factor AP-2 Transcription Factors proenkephalin cytidylyl-3'-5'-guanosine 5-Methylcytosine Cytosine DNA Deoxyribonuclease HpaII Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Comb M
Department of Genetics, Harvard Medical School, Boston, MA 02114.
Goodman H M
References (36)
36 references, click to expand
  1. Proteins bound at adjacent DNA elements act synergistically to regulate human proenkephalin cAMP inducible transcription.
    EMBO J. 1988 Dec 1;7(12):3793-805 PMID: 2850173
  2. Positive and negative regulation of transcription in vitro: enhancer-binding protein AP-2 is inhibited by SV40 T antigen.
    Cell. 1987 Sep 11;50(6):847-61 PMID: 3040262
  3. Role of an adenovirus E2 promoter binding factor in E1A-mediated coordinate gene control.
    Proc Natl Acad Sci U S A. 1987 Apr;84(8):2180-4 PMID: 2951737
  4. The coupling between enhancer activity and hypomethylation of kappa immunoglobulin genes is developmentally regulated.
    Mol Cell Biol. 1988 Feb;8(2):930-7 PMID: 3127693
  5. Multiple nuclear factors interact with the immunoglobulin enhancer sequences.
    Cell. 1986 Aug 29;46(5):705-16 PMID: 3091258
  6. Cytosine methylation in CTF and Sp1 recognition sites of an HSV tk promoter: effects on transcription in vivo and on factor binding in vitro.
    Nucleic Acids Res. 1989 Dec 25;17(24):10179-90 PMID: 2557588
  7. CpG methylation of the cAMP-responsive enhancer/promoter sequence TGACGTCA abolishes specific factor binding as well as transcriptional activation.
    Genes Dev. 1989 May;3(5):612-9 PMID: 2545524
  8. An AP-2 element acts synergistically with the cyclic AMP- and phorbol ester-inducible enhancer of the human proenkephalin gene.
    Mol Cell Biol. 1989 Jan;9(1):321-4 PMID: 2538722
  9. A common trans-acting factor is involved in transcriptional regulation of neurotransmitter genes by cyclic AMP.
    Mol Cell Biol. 1988 Oct;8(10):4225-33 PMID: 2903436
  10. Genomic footprinting reveals cell type-specific DNA binding of ubiquitous factors.
    Cell. 1987 Nov 6;51(3):435-43 PMID: 2889531
  11. DNA methylation and gene activity.
    Cell. 1988 Apr 8;53(1):3-4 PMID: 3280142
  12. Cytosine methylation does not affect binding of transcription factor Sp1.
    Proc Natl Acad Sci U S A. 1988 Apr;85(7):2066-70 PMID: 3281160
  13. Regulation of inducible and tissue-specific gene expression.
    Science. 1987 Jun 5;236(4806):1237-45 PMID: 3296191
  14. 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):2827-31 PMID: 3458243
  15. 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):7167-71 PMID: 3463957
  16. Affinity purification of sequence-specific DNA binding proteins.
    Proc Natl Acad Sci U S A. 1986 Aug;83(16):5889-93 PMID: 3461465
  17. Site-specific demethylation and normal chromatin structure of the human dihydrofolate reductase gene promoter after transfection into CHO cells.
    Mol Cell Biol. 1987 Aug;7(8):2830-7 PMID: 3670295
  18. DNA methylation and the regulation of globin gene expression.
    Cell. 1983 Aug;34(1):197-206 PMID: 6883509
  19. 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
  20. Expression of the chloramphenicol acetyltransferase gene in mammalian cells under the control of adenovirus type 12 promoters: effect of promoter methylation on gene expression.
    Proc Natl Acad Sci U S A. 1983 Dec;80(24):7586-90 PMID: 6324179
  21. Primary structure of the human proenkephalin gene.
    DNA. 1983;2(3):213-29 PMID: 6315326
  22. High efficiency DNA-mediated transformation of primate cells.
    Science. 1983 Aug 5;221(4610):551-3 PMID: 6306768
  23. DNA methylation of three 5' C-C-G-G 3' sites in the promoter and 5' region inactivate the E2a gene of adenovirus type 2.
    Proc Natl Acad Sci U S A. 1984 May;81(10):2950-4 PMID: 6328479
  24. 5-Methylcytosine in eukaryotic DNA.
    Science. 1981 Jun 19;212(4501):1350-7 PMID: 6262918
  25. DNA methylation and gene function.
    Science. 1980 Nov 7;210(4470):604-10 PMID: 6254144
  26. CpG-rich islands and the function of DNA methylation.
    Nature. 1986 May 15-21;321(6067):209-13 PMID: 2423876
  27. DNA methylation affects the formation of active chromatin.
    Cell. 1986 Feb 28;44(4):535-43 PMID: 3456276
  28. Site specific demethylation in the promoter of human gamma-globin gene does not alleviate methylation mediated suppression.
    EMBO J. 1987 Aug;6(8):2329-35 PMID: 3665878
  29. A fraction of the mouse genome that is derived from islands of nonmethylated, CpG-rich DNA.
    Cell. 1985 Jan;40(1):91-9 PMID: 2981636
  30. A simple phase-extraction assay for chloramphenicol acyltransferase activity.
    Gene. 1988 Jul 30;67(2):271-7 PMID: 3169576
  31. Transcription factor AP-2 mediates induction by two different signal-transduction pathways: protein kinase C and cAMP.
    Cell. 1987 Oct 23;51(2):251-60 PMID: 2822255
  32. A cyclic AMP- and phorbol ester-inducible DNA element.
    Nature. 1986 Sep 25-Oct 1;323(6086):353-6 PMID: 3020428
  33. Cell-specific expression of the rat insulin gene: evidence for role of two distinct 5' flanking elements.
    Science. 1985 Nov 22;230(4728):912-6 PMID: 3904002
  34. Trans effect of the E1 region of adenoviruses on the expression of a prokaryotic gene in mammalian cells: resistance to 5' -CCGG- 3' methylation.
    Proc Natl Acad Sci U S A. 1986 Mar;83(6):1598-1602 PMID: 2937060
  35. Methylation of single CpG dinucleotides within a promoter element of the Herpes simplex virus tk gene reduces its transcription in vivo.
    Gene. 1988 May 30;65(2):219-27 PMID: 2842233
  36. A cellular protein, activating transcription factor, activates transcription of multiple E1A-inducible adenovirus early promoters.
    Proc Natl Acad Sci U S A. 1987 Dec;84(23):8355-9 PMID: 2960975
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/18.13.3975
Published
1990-07-11
Pages
3975-82
Language
English
Region
England
NLM ID
0411011
PMCID
PMC331101
Subset
IM
Grants
NIDA NIH HHS · R01-DA05706-01 · United States
External Links
PubMed source DOI: 10.1093/nar/18.13.3975 PMC Full Text View journal details
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