A region upstream of the human delta-globin gene shows a stage-specific interaction with globin promoters in erythroid cell lines.
We previously showed that the 651-bp DNA fragment, located 3 kb upstream from the human delta-globin gene (fragment F5), is able to inhibit adult, not fetal, globin promoter in mouse erythroleukemia cell lines (MEL) expressing adult globin genes. Here we show in transient analysis that fragment F5 has a strong inhibitory effect on fetal gamma-globin promoter in human erythroleukemia cell lines (HEL) expressing fetal globin genes. Since the beta-promoter constructs were poorly expressed in fetal cells, new plasmids containing an HPFH promoter (Ggamma(-175), T to C), which is strongly expressed in both fetal and adult cell lines, were made. Here we report that fragment F5 in HEL cells has a strong inhibitory effect on wild-type gamma-promoter only; no effect was evident on gamma(-175)-promoter in either MEL or HEL cell lines. Altogether these results show a stage-specific interaction between fragment F5 and globin promoters during development. We also report the presence of several bindings for erythroid GATA family factors by electrophoretic mobility shift assay, using nuclear extracts from erythroid cell lines.
Vitale
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Unità di Ricerca Piera Cutino, Divisione di Ematologia II, Ospedale V. Cervello, via Trabucco, n. 180, Palermo, 90146, Italy. amaggio@skyol.it
Calzolari
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Di Marzo
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Acuto
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Maggio
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A
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000378:metabolism; D015672:Erythroid Precursor Cells / Q000378:metabolism*; D005326:Fetal Proteins / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D005914:Globins / Q000235:genetics*; D006801:Humans; D051379:Mice; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured
D004268:DNA-Binding Proteins; D005326:Fetal Proteins; D005914:Globins