Binding of the transcription factor EBP-80 mediates the methylation response of an intracisternal A-particle long terminal repeat promoter.
Open accessAbstract
Intracisternal A-particle (IAP) expression in mouse cells has been correlated with hypomethylation of HhaI and HpaII sites in proviral long terminal repeats (LTRs). In a previous study, in vitro methylation of three HhaI sites in the U3 region of the LTR from the cloned genomic IAP element, MIA14, was shown to inhibit promoter activity in vivo. In this study, we found by site-directed mutagenesis that the two more downstream HhaI sites within this LTR were responsible for the methylation effects on promoter activity in vivo; methylation of the other (5') HhaI site, which lies within a putative SP1 binding domain, did not affect promoter activity. Methylation of the HhaI sites also inhibited promoter activity of the LTR in a cell-free transcription system. Exonuclease III footprinting demonstrated methylation-induced changes in protein binding over the region encompassing the downstream HhaI site, designated the Enh2 domain. The protein that interacts specifically with this domain, EBP-80, was characterized in a previous study (M. Falzon and E. L. Kuff, J. Biol. Chem. 264:21915-21922, 1989). We show here that the presence of methylcytosine in the HhaI site within the Enh2 domain inhibited binding of EBP-80 in vitro. The methylated MIA14 LTR construct was much less responsive to added EBP-80 in an in vitro transcription system than was the unmethylated construct. These data suggest that CpG methylation within the Enh2 domain may exert its effect on transcription in vivo by altering the interaction between EBP-80 and its cognate DNA sequence.
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Manipulation of the methylation status of DNA sequences results in differential binding of transcription factors, including E2F, AP2α, MYC and MYN.
“The manipulation of the methylation status of DNA sequences has been shown (mostly in in vitro studies) to result in the differential binding of TFs, including E2F, AP2α, MYC and MYN88–95.”
From Transcription factors as readers and effectors of DNA methylation (2016) · cites this paper as [91]
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- Authors
- Falzon, M., & Kuff, E. L.
- Affiliations
- National Institutes of Health (US)
- Venue
- Molecular and Cellular Biology
- Publisher
- Informa UK Limited
- Published
- 1991-01
- Volume
- 11
- Issue
- 1
- Pages
- 117-125
- ISSN
- 0270-7306 (ISSN-L)
- 1098-5549
- PMC
- PMC359600
- OpenAlex
- W1804699878
- Cited by
- 37
- References
- 46
- Citations by year
- 2024: 1 · 2022: 1 · 2020: 1 · 2016: 1 · 2012: 1
- FWCI
- 1.98
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Molecular and Cellular Biology (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (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
- intracisternal A-particle, long terminal repeat promoter, CpG methylation, DNA methylation, transcription factor binding, promoter activity, protein-DNA interactions, DNA hypomethylation
- MeSH terms
- Base Sequence, Binding Sites, Cell Line, DNA-Binding Proteins, Humans, Genes, Intracisternal A-Particle, Methylation, Molecular Sequence Data, Promoter Regions, Genetic, Protein Binding, Regulatory Sequences, Nucleic Acid, Repetitive Sequences, Nucleic Acid, Transcription Factors, Transcription, Genetic, Restriction Mapping, In Vitro Techniques
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