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

FEBS Letters

Schaffner, Walter¹  Georgiev, Oleg¹  Matsuo, Koichi¹  Silke, John¹  Rother, Kristina I.¹ 

[1] Institut für Molekularbiologie II, Universität Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland

关键词: DNA methylation;    Transcription factor;    Bisulphite PCR;    EC;    embryonal carcinoma;    ES;    embryonic stem cells;    aprt;    adenine phosphoribosyl transferase;    Hprt;    hypoxanthine phosphoribosyl transferase;   

DOI  :  10.1016/0014-5793(95)00830-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown