DNA methylation at a CCGG sequence in the large intron of the rabbit β-globin gene: tissue-specific variations

Abstract

We have analysed DNA modification in a HapII site (CCGG) present in the major intron of the discontinuous rabbit beta-globin gene. In most somatic tissues, including erythroid and non-erythroid tissues, about 50% of the DNA is resistant to cleavage at this site by HapII, though 100% cleavage is found with the isoschizomer MspI. Since the former enzyme is unable to cleave CCGG sites if the internal C residue is 5-methyl C (and since methylation is the only form of CpG modification documented in animal DNA), while the latter enzyme cleaves DNA irrespective of methylation at this residue, we infer that 50% of the CCGG sites in the beta-globin gene intron are methylated in these tissues. The same site appears to be 100% methylated (judged by the same criterium) in sperm DNA and about 80% methylated in brain DNA. DNA from the rabbit SIRC cell line is entirely unmethylated at this site.