DOI: 10.1101/gad.3.5.612. CpG methylation of the cAMP-responsive enhancer/promoter sequence TGACGTCA abolishes specific factor binding as well as transcriptional activation.

CpG methylation of the cAMP-responsive enhancer/promoter sequence TGACGTCA abolishes specific factor binding as well as transcriptional activation.

10.1101/gad.3.5.612

Crossref journal-article

Cold Spring Harbor Laboratory

Genes & Development (246)

Abstract

In mammals and other vertebrates, cytosine methylation in CpG sites is often negatively correlated with gene activity. Because methylation of the promoter region is most crucial for this effect, the simplest hypothesis is that CpG methylation interferes with the binding of specific transcription factors. We have examined this hypothesis with two different transcription factor-binding sites that contain a CpG dinucleotide, namely the cAMP-responsive element (CRE; 5'-TGACGTCA) and the Sp1-binding site (5'-GTGAGGCGGTGAGACT). We have reported previously that CpG methylation of the Sp1-binding site affected neither factor binding nor transcription in HeLa cells, which may be related to the fact that Sp1 is typically associated with promoters of housekeeping genes. In contrast, CREs are often associated with promoters of cell type-specific genes. A synthetic oligonucleotide containing two tandem CREs derived from the gene encoding the human glycoprotein hormone alpha-subunit was cloned upstream of a reporter gene. Transcription of this gene was dependent on the CRE sequences in both PC12 and HeLa cells. Bandshift and methylation interference assays show that similar, if not the same, factor(s) bind to the CRE in both cell lines, even though induction by cAMP was only observed in PC12 cells. CpG methylation of the CRE consensus sequences (TGACGTCA) resulted in loss of specific factor binding, as well as loss of transcriptional activity in vitro and in vivo, in both cell types. This suggests that the inactivity of methylated promoters can, at least in some cases, be explained by their inability to bind specific transcription factors.

Bibliography

Iguchi-Ariga, S. M., & Schaffner, W. (1989). CpG methylation of the cAMP-responsive enhancer/promoter sequence TGACGTCA abolishes specific factor binding as well as transcriptional activation. Genes & Development, 3(5), 612â619.

Authors 2

S M Iguchi-Ariga (first)
W Schaffner (additional)

References 35 Referenced 429

10.1016/0092-8674(87)90639-8
10.1016/0378-1119(88)90458-1
10.1038/321209a0
{'key': '2021111319552323000_3.5.612.4', 'first-page': '324', 'article-title': 'CpG islands as gene markers in the vertebrate nucleus.', 'volume': '3', 'year': '1987', 'journal-title': 'Trends Genet.'} / Trends Genet. / CpG islands as gene markers in the vertebrate nucleus. (1987)
10.1073/pnas.84.5.1177
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Cristy, B.A. and G.A. Scangos. 1986. Control of gene expression by DNA methylation. In Molecular genetics of mammalian cells (ed. G.M. Malacinski), pp. 3–29, Macmillan, New York.
{'key': '2021111319552323000_3.5.612.9', 'first-page': '3994', 'article-title': 'Tissue-specific enhancer of the human glycoprotein hormone α-subunit gene: Dependence on cyclic AMP-inducible elements.', 'volume': '7', 'year': '1987', 'journal-title': 'Mol. Cell Biol.'} / Mol. Cell Biol. / Tissue-specific enhancer of the human glycoprotein hormone α-subunit gene: Dependence on cyclic AMP-inducible elements. (1987)
de Villiers, J. and W. Schaffner. 1983. Transcriptional ‘enhancers’ from papovaviruses as components of eukaryotic expression vectors. In Techniques in the life sciences, B5. Nucleic Acid Biochemistry, Elsevier, Ireland.
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{'key': '2021111319552323000_3.5.612.19', 'first-page': '3032', 'article-title': 'trans-Acting factors interact with a cyclic AMP response element to modulate expression of the human gonadotropin gene.', 'volume': '7', 'year': '1987', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / trans-Acting factors interact with a cyclic AMP response element to modulate expression of the human gonadotropin gene. (1987)
{'key': '2021111319552323000_3.5.612.20', 'first-page': '930', 'article-title': 'The coupling between enhancer activity and hypomethylation of α-immunoglobulin genes is developmentally regulated.', 'volume': '8', 'year': '1988', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / The coupling between enhancer activity and hypomethylation of α-immunoglobulin genes is developmentally regulated. (1988)
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10.1073/pnas.84.8.2180
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10.1016/S0021-9258(18)69229-4 / J. Biol. Chem. / Transcriptional regulation of a plasminogen activator gene by cyclic AMP in a homologous cell-free system. (1988)
Parker, J.-H., H.V. Hershey, and M.W. Taylor. 1986. House-keeping genes. In Molecular genetics of mammalian cells (ed. G.M. Malacinski), pp. 79–98, Macmillan, New York.
10.1016/S0021-9258(19)76506-5 / J. Biol. Chem. / Cyclic AMP and the induction of eukaryotic gene transcription. (1988)
10.1073/pnas.83.19.7167
{'key': '2021111319552323000_3.5.612.29', 'first-page': '2830', 'article-title': 'Site-specific demethylation and normal chromatin structure of the human dihydrofolate reductase gene promoter after transfection into CHO cells.', 'volume': '7', 'year': '1987', 'journal-title': 'Mol. Cell Biol.'} / Mol. Cell Biol. / Site-specific demethylation and normal chromatin structure of the human dihydrofolate reductase gene promoter after transfection into CHO cells. (1987)
10.1073/pnas.77.1.122
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10.1093/nar/15.17.6787
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10.1093/nar/5.4.1139

Dates

Type	When
Created	18 years, 2 months ago (June 5, 2007, 5:15 p.m.)
Deposited	3 years, 9 months ago (Nov. 13, 2021, 10:57 p.m.)
Indexed	1 month ago (July 28, 2025, 2:30 a.m.)
Issued	36 years, 3 months ago (May 1, 1989)
Published	36 years, 3 months ago (May 1, 1989)
Published Online	36 years, 3 months ago (May 1, 1989)
Published Print	36 years, 3 months ago (May 1, 1989)

Funders 0

None

BibTeX

@article{Iguchi_Ariga_1989, title={CpG methylation of the cAMP-responsive enhancer/promoter sequence TGACGTCA abolishes specific factor binding as well as transcriptional activation.}, volume={3}, ISSN={1549-5477}, url={http://dx.doi.org/10.1101/gad.3.5.612}, DOI={10.1101/gad.3.5.612}, number={5}, journal={Genes & Development}, publisher={Cold Spring Harbor Laboratory}, author={Iguchi-Ariga, S M and Schaffner, W}, year={1989}, month=may, pages={612–619} }

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