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Complex demethylation patterns at Sp1 binding sites in F9 embryonal carcinoma cells
Author(s) -
Silke John,
Rother Kristina I.,
Georgiev Oleg,
Schaffner Walter,
Matsuo Koichi
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)00830-3
Subject(s) - dna methylation , cpg site , methylation , demethylation , dna demethylation , microbiology and biotechnology , biology , dna , bisulfite sequencing , illumina methylation assay , dna sequencing , differentially methylated regions , dna binding site , plasmid , promoter , chemistry , genetics , gene , gene expression
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.