Methylation-sensitive protein-DNA interaction at the cell cycle regulatory domain of human thymidine kinase promoter

We have investigated the DNA-protein complex formation in nuclear extracts of human cells using the sequence of cell-cycle regulatory unit (CCRU) of human thymidine kinase (TK) promoter. It appeared that a distinct DNA-protein complex was present in three human tumor cell lines and that the CCAAT box within the sequence of CCRU was a necessary element for complex formation. Upon 4 days of serum deprivation, this DNA-protein complex remained unchanged in HeLa cells, but the expression of TK mRNA was decreased. Furthermore, DNA methylation of the Hhal site of the CCRU sequence of the TK promoter greatly reduced the binding activity of nuclear proteins from different human tumor cell lines. On the basis of these data, we proposed a possible role for DNA methylation in the regulation of TK transcription during late G1/S phase progression of the cell cycle.