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 journalArticle

25 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 languageEnglish
Pages (from-to)170-174
Number of pages5
JournalFEBS Letters
Volume370
Issue number3
DOIs
Publication statusPublished - 1995 Aug 21
Externally publishedYes

Fingerprint

Embryonal Carcinoma Stem Cells
Methylation
Binding Sites
Cells
DNA
Sp1 Transcription Factor
CpG Islands
Plasmids
Molecules

Keywords

  • Bisulphite PCR
  • DNA methylation
  • Transcription factor

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Genetics
  • Molecular Biology
  • Structural Biology

Cite this

Complex demethylation patterns at Sp1 binding sites in F9 embryonal carcinoma cells. / Silke, John; Rother, Kristina I.; Georgiev, Oleg; Schaffner, Walter; Matsuo, Koichi.

In: FEBS Letters, Vol. 370, No. 3, 21.08.1995, p. 170-174.

Research output: Contribution to journalArticle

Silke, John ; Rother, Kristina I. ; Georgiev, Oleg ; Schaffner, Walter ; Matsuo, Koichi. / Complex demethylation patterns at Sp1 binding sites in F9 embryonal carcinoma cells. In: FEBS Letters. 1995 ; Vol. 370, No. 3. pp. 170-174.
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