CpG methylation of half-CRE sequences creates C/EBPα binding sites that activate some tissue-specific genes

V Rishi, P Bhattacharya, R Chatterjee… - Proceedings of the …, 2010 - National Acad Sciences
Proceedings of the National Academy of Sciences, 2010National Acad Sciences
DNA methylation of the cytosine in the CpG dinucleotide is typically associated with gene
silencing. Genomic analyses have identified low CpG promoters that are both methylated
and transcriptionally active, but the mechanism underlying the activation of these methylated
promoters remains unclear. Here we show that CpG methylation of the CRE sequence
(TGACGTCA) enhances the DNA binding of the C/EBPα transcription factor, a protein critical
for activation of differentiation in various cell types. Transfection assays also show that …
DNA methylation of the cytosine in the CpG dinucleotide is typically associated with gene silencing. Genomic analyses have identified low CpG promoters that are both methylated and transcriptionally active, but the mechanism underlying the activation of these methylated promoters remains unclear. Here we show that CpG methylation of the CRE sequence (TGACGTCA) enhances the DNA binding of the C/EBPα transcription factor, a protein critical for activation of differentiation in various cell types. Transfection assays also show that C/EBPα activates the CRE sequence only when it is methylated. The biological significance of this observation was seen in differentiating primary keratinocyte cultures from newborn mice where certain methylated promoters are both bound by C/EBPα and activated upon differentiation. Experimental demethylation by either 5-azacytidine treatment or DNMT1 depletion diminished both C/EBPα binding and activation of the same methylated promoters upon differentiation suggesting that CpG methylation can localize C/EBPα. Transfection studies in cell cultures using methylated tissue-specific proximal promoters identified half-CRE (CGTCA) and half-C/EBP (CGCAA) sequences that need to be methylated for C/EBPα mediated activation. In primary dermal fibroblasts, C/EBPα activates a different set of methylated tissue-specific promoters upon differentiation into adipocytes. These data identify a new function for methyl CpGs: producing DNA binding sites at half-CRE and half-C/EBP sequences for C/EBPα that are needed to activate tissue-specific genes.
National Acad Sciences