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Hypomethylation of DNA derived from purified human erythroid cells correlates with gene activity of the beta-globin cluster

A Oppenheim, Y Katzir, E Fibach, A Goldfarb and E Rachmilewitz

Analysis of methylation at the beta-globin gene cluster was carried out on DNA derived from nucleated RBCs (orthochromatic normoblasts) isolated from peripheral blood of patients with beta-thalassemia major or other congenital hemolytic anemia after splenectomy. A procedure to separate these normoblasts from the other nucleated cells of the peripheral blood was developed, providing us with a convenient source of DNA for investigating parameters related to human erythroid differentiation. Blood samples were obtained from six adult patients who express their gamma-globin genes at different levels. Inverse correlation between methylation and gene activity was consistently observed for five of the eight sites analyzed. A site 3' to the beta gene was always unmethylated, two sites flanking the epsilon gene were always found to be methylated, and two sites 5' to the two gamma genes, G gamma and A gamma, were hypomethylated in correlation with gamma gene activity of the individual patients. A site 5' to the delta gene was unmethylated in normoblasts as well as in WBC. No apparent relation between hypomethylation and gene activity was observed for two additional sites. The results suggest that methylation at specific chromosomal locations participate in genetic regulation of the beta- like globin genes in humans.

Volume 66, Issue 5, pp. 1202-1207, 11/01/1985
Copyright © 1985 by The American Society of Hematology


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M. Hsu, R. Mabaera, C. H. Lowrey, D. I. K. Martin, and S. Fiering
CpG Hypomethylation in a Large Domain Encompassing the Embryonic {beta}-Like Globin Genes in Primitive Erythrocytes
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