Nuclease hypersensitivity in the beta-globin gene region of K562 cells.
We have investigated chromatin structure in the beta-globin gene region of the K562 human erythroleukemic cell line by using S1 and DNase I nuclease sensitivity assays. Despite the lack of beta-globin gene expression in these cells, we find nuclease-hypersensitive sites to these enzymes in its 5' and 3' flanking regions in K562 chromatin. This result is in contrast to previous reports in which no hypersensitive sites were found in the immediate vicinity of this gene. In the 3' region, one major hypersensitive site at 0.9 kpb 3' and three minor hypersensitive sites at 0.7 kbp, 0.5 kbp 3' and 0.2 kbp 5' of the polyadenylation site were observed; these sites are very similar to those found in fetal liver and adult bone marrow cells in which the beta-globin gene is expressed. We find hypersensitive sites to both enzymes in the 5' region of the beta-globin gene: a major site 0.8 kbp 5' to the cap site, and two minor sites 1.2 and 1.5 kbp 5' to the cap site. The -0.8 kbp site is also present in plasmids containing the beta-globin gene. Our results suggest that the lack of beta-globin gene expression may be related to the lack of hypersensitivity sites in the immediate (150 bp) 5' flanking region of the beta-globin gene, as occurs in other active globin genes.
D000328:Adult; D001665:Binding Sites; D002460:Cell Line; D002467:Cell Nucleus / Q000032:analysis; D002843:Chromatin / Q000032:analysis*; D004247:DNA / Q000032:analysis; D003850:Deoxyribonuclease I; D004720:Endonucleases; D005796:Genes; D005914:Globins / Q000235:genetics*; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute; D009710:Nucleotide Mapping; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012313:RNA / Q000032:analysis; D015719:Single-Strand Specific DNA and RNA Endonucleases; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured / Q000378:metabolism
D002843:Chromatin; D012313:RNA; D005914:Globins; D004247:DNA; D004720:Endonucleases; D003850:Deoxyribonuclease I; D015719:Single-Strand Specific DNA and RNA Endonucleases