title: Undermethylation and DNase I hypersensitivity of myeloperoxidase gene in HL-60 cells before and after differentiation.
abstract: Methylation and DNase I-hypersensitive sites of the myeloperoxidase gene in human myeloid leukemia HL-60 cells were studied by Southern blot hybridization using the myeloperoxidase gene probes. Digestion of DNA with a methylation-sensitive restriction endonuclease indicated that a CpG in the CCGG sequence located 3.53 kbp upstream of the myeloperoxidase gene was unmethylated in HL-60 cells expressing the gene, whereas it was methylated in K562 cells and human placenta not expressing the gene. The site in HL-60 cells remained unmethylated after retinoic acid- or 12-O-tetradecanoyl-phorbol-13-acetate-induced differentiation that arrests myeloperoxidase synthesis. Digestion of isolated nuclei with various amounts of DNase I indicated that four DNase I-hypersensitive sites were in an upstream region of the myeloperoxidase gene in HL-60 cells and three sites were within the gene. In retinoic acid-induced cells, the bands of the hypersensitive site near the 5' side of the gene and that in the first intron became weak, while that of the site in the fifth intron became strong. The bands of these hypersensitive sites were weak in K562 cells. The implications of these changes in tissue-specific expression and developmental down-regulation of the myeloperoxidase gene are discussed.
authors.affiliation: Institute for Protein Research, Osaka University, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Hashinaka
authors.forename: M
authors.initials: M
authors.lastname: Yamada
mesh_terms: D001483:Base Sequence; D002454:Cell Differentiation / Q000187:drug effects; D002843:Chromatin / Q000648:ultrastructure; D003850:Deoxyribonuclease I / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic; D005796:Genes; D006801:Humans; D007951:Leukemia, Myeloid / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D008745:Methylation; D008969:Molecular Sequence Data; D009195:Peroxidase / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured
chemical_list: D002843:Chromatin; D014212:Tretinoin; D009195:Peroxidase; D003850:Deoxyribonuclease I