Murine erythroleukemia cell differentiation: DNase I hypersensitivity and DNA methylation near the globin genes

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The sensitivity to digestion by DNase I of chromatin containing the α- and βmajor-globin genes and the pattern of DNA methylation near these genes was examined during hexamethylenebisacetamide (HMBA)-mediated erythroid differentiation of murine erythroleukemia cells (MELC). In uninduced and induced cells, the chromatin regions containing the α- and β-major-globin genes are more sensitive to digestion by DNase I than is the region containing an immunoglobulin gene (Igα) not expressed during erythroid differentiation. However, at low concentrations of DNase I, a 6- to 10-fold increase in site-specific cleavages was generated in chromatin regions near both the α- and βmajor-globin genes in cells induced to differentiate by HMBA. The DNase I hypersensitive site near the βmajor-globin gene maps to a small region near the 5′ terminus of the gene. No detectable change in the pattern of DNA methylation around either the α- or β-globin genes was observed during HMBA-mediated erythroid differentiation. Of the potentially methylated sites assayed and mapped near the βmajor-globin gene, one site is fully methylated, one is partially methylated, and one is unmethylated both in uninduced and induced cells. Many (but not all) sites assayed near the α-globin genes are unmethylated in both uninduced and induced cells. These results show that specific alterations of chromatin structure occur during MELC differentiation and suggest that these changes may not involve alterations in the pattern of DNA methylation.

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