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 | |
| Publication status | Published - 1995 Aug 21 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Complex demethylation patterns at Sp1 binding sites in F9 embryonal carcinoma cells
AU - Silke, John
AU - Rother, Kristina I.
AU - Georgiev, Oleg
AU - Schaffner, Walter
AU - Matsuo, Koichi
PY - 1995/8/21
Y1 - 1995/8/21
N2 - 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.
AB - 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.
KW - Bisulphite PCR
KW - DNA methylation
KW - Transcription factor
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U2 - 10.1016/0014-5793(95)00830-3
DO - 10.1016/0014-5793(95)00830-3
M3 - Article
VL - 370
SP - 170
EP - 174
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 3
ER -