PreprintCited by 559Epigenetics and DNA MethylationGenomics and Chromatin DynamicsCancer-related gene regulation

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

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Iguchi-Ariga, S M, Schaffner, W·1989·Genes & Development

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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.

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Authors: Iguchi-Ariga, S. M., & Schaffner, W.
Affiliations: University of Zurich (CH)
Venue: Genes & Development
Publisher: Cold Spring Harbor Laboratory
Published: 1989-05
Volume: 3
Issue: 5
Pages: 612-619
ISSN: 0890-9369 (ISSN-L)
1549-5477
DOI: 10.1101/gad.3.5.612
OpenAlex: W2153782423
Cited by: 559
References: 32
Citations by year: 2026: 3 · 2025: 9 · 2024: 10 · 2023: 11 · 2022: 14 · 2021: 11 · 2020: 11 · 2019: 15 · 2018: 12 · 2017: 15 · 2016: 18 · 2015: 14 · 2014: 20 · 2013: 19 · 2012: 20
Type: posted content
Language: en
Open access: bronze
doi.org
Locations: Genes & Development (publishedVersion)
PubMed (publishedVersion)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cancer-related gene regulation
Keywords: CpG methylation, cAMP response element, Sp1 binding site, transcription factor binding, transcriptional activation, promoter methylation, gene silencing, glycoprotein hormone alpha-subunit, PC12 cells, HeLa cells, methylation interference, cAMP signaling
MeSH terms: Cyclic AMP, Base Sequence, Binding Sites, Cells, Cultured, DNA, DNA-Binding Proteins, Enhancer Elements, Genetic, HeLa Cells, Humans, Methylation, Molecular Sequence Data, Promoter Regions, Genetic, Transcription, Genetic, Dinucleoside Phosphates

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

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