NF‐E2 and GATA binding motifs are required for the formation of DNase I hypersensitive site 4 of the human beta‐globin locus control region.

The beta‐like globin genes require the upstream locus control region (LCR) for proper expression. The active elements of the LCR coincide with strong erythroid‐specific DNase I‐hypersensitive sites (HSs). We have used 5′ HS4 as a model to study the formation of these HSs. Previously, we identified a 101 bp element that is required for the formation of this HS. This element binds six proteins in vitro. We now report a mutational analysis of the HS4 HS‐forming element (HSFE). This analysis indicates that binding sites for the hematopoietic transcription factors NF‐E2 and GATA‐1 are required for the formation of the characteristic chromatin structure of the HS following stable transfection into murine erythroleukemia cells. Similarly arranged NF‐E2 and GATA binding sites are present in the other HSs of the human LCR, as well as in the homologous mouse and goat sequences and the chicken beta‐globin enhancer. A combination of DNase I and micrococcal nuclease sensitivity assays indicates that the characteristic erythroid‐specific hypersensitivity of HS4 to DNase I is the result of tissue‐specific alterations in both nucleosome positioning and tertiary DNA structure.