Journal
MOLECULAR BIOLOGY REPORTS
Volume 40, Issue 2, Pages 797-801Publisher
SPRINGER
DOI: 10.1007/s11033-012-2117-7
Keywords
CpG island; GATA-2; C/EBP beta; Preadipocyte; Methylation
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Funding
- Committee of Science and Technology of Shanghai, China [02DJ14005]
- 211 key program
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Mammalian genomes are punctuated by DNA sequences containing an atypically high frequency of CpG sites (CpG islands; CGIs) that are associated with the majority of annotated gene promoters. Methylated C bases of CpG sites inhibit the expression of downstream genes. During the differentiation of 3T3-L1 preadipocytes, the CCAAT/enhancer-binding protein (C/EBP) beta gene plays an important role. We studied the CpG island methylation status of the C/EBP beta promoter and its relationship with the GATA-2 protein. We used computer analysis to determine that the C/EBP beta promoter sequence is rich in CGIs, and observed that two of seven methylated C bases were demethylated during the preadipocyte differentiation using bisulfite sequencing PCR (BSP). This corresponded with the onset of notable C/EBP beta gene expression. Immunofluorescence and molecular docking showed that the GATA-2 protein binds the C/EBP beta promoter in front of the first demethylated CpG site. We also found that expression of GATA-2 and C/EBP beta proteins is negatively correlated. These results indicate that the methylated C bases in the C/EBP beta promoter relate to expression of the C/EBP beta gene, and that its demethylation is linked with GATA-2 protein association.
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