Complex demethylation patterns at Sp1 binding sites in F9 embryonal carcinoma cells

John Silke; Kristina I. Rother; Oleg Georgiev; Walter Schaffner; Koichi Matsuo

doi:10.1016/0014-5793(95)00830-3

Complex demethylation patterns at Sp1 binding sites in F9 embryonal carcinoma cells

John Silke, Kristina I. Rother, Oleg Georgiev, Walter Schaffner, Koichi Matsuo

Research output: Contribution to journal › Article › peer-review

26 Citations (Scopus)

Abstract

The ubiquitous transcription factor Sp1 has been implicated in the mechanism which maintains CpG islands methylation-free. Plasmids containing GC boxes (Sp1 sites) were in vitro methylated at every CpG dinucleotide. After stable introduction into F9 embryonal carcinoma cells, we analysed the methylation of the sequence around the GC boxes with bisulphite sequencing. In agreement with restriction site analysis by other labs, we found preferential demethylation at GC box DNA versus control DNA. However, the bisulphite sequencing which permits the analysis of every CpG site on a given DNA molecule, revealed a complex pattern of methylated and unmethylated sites. Upon prolonged culture the pattern became simpler, with most sites demethylated but certain sites being consistently methylated.

Original language	English
Pages (from-to)	170-174
Number of pages	5
Journal	FEBS Letters
Volume	370
Issue number	3
DOIs	https://doi.org/10.1016/0014-5793(95)00830-3
Publication status	Published - 1995 Aug 21
Externally published	Yes

Keywords

Bisulphite PCR
DNA methylation
Transcription factor

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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abstract = "The ubiquitous transcription factor Sp1 has been implicated in the mechanism which maintains CpG islands methylation-free. Plasmids containing GC boxes (Sp1 sites) were in vitro methylated at every CpG dinucleotide. After stable introduction into F9 embryonal carcinoma cells, we analysed the methylation of the sequence around the GC boxes with bisulphite sequencing. In agreement with restriction site analysis by other labs, we found preferential demethylation at GC box DNA versus control DNA. However, the bisulphite sequencing which permits the analysis of every CpG site on a given DNA molecule, revealed a complex pattern of methylated and unmethylated sites. Upon prolonged culture the pattern became simpler, with most sites demethylated but certain sites being consistently methylated.",

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AU - Silke, John

AU - Rother, Kristina I.

AU - Georgiev, Oleg

AU - Schaffner, Walter

AU - Matsuo, Koichi

PY - 1995/8/21

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Complex demethylation patterns at Sp1 binding sites in F9 embryonal carcinoma cells

Abstract

Keywords

ASJC Scopus subject areas

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