In this paper we describe a complete deletional analysis of hypersensitive site three (HS3) of the human beta-globin Locus Control Region (LCR). The previously defined core fragment consists of 6 footprinted regions, with multiple binding sites for the erythroid-specific factor GATA-1 and G-rich motifs that can interact with ubiquitous factors such as Sp1 and TEF-2. We show in this paper that the 5' half of this fragment is the most important for activity in murine erythroleukemia (MEL) cells. A fragment containing footprints 1-4 can stimulate transcription of a linked human beta-globin gene to levels of about 40% of that obtained with footprints 1-6. Constructs containing either footprints 1-3 or 3-6 cannot be distinguished from the beta-globin gene alone. We further show that binding sites for the erythroid-specific factor NF-E2 can co-operatively interact with parts of the HS3 core fragment, and that HS3 requires elements upstream from -103 in the human beta-globin promoter for full activity. The importance of these results is discussed in the context of the regulation of the genes in the human beta-globin cluster.

*Gene Expression Regulation, *Promoter Regions (Genetics), *Transcription, Genetic, 0 (DNA Primers), 0 (DNA-Binding Proteins), 0 (Transcription Factors), 125267-48-3 (erythroid-specific DNA-binding factor), 9004-22-2 (Globins), Animals, Base Sequence, Binding Sites, DNA Primers/chemistry, DNA-Binding Proteins/metabolism, Globins/*genetics, Human, In Vitro, Leukemia, Erythroblastic, Acute, Mice, Molecular Sequence Data, Sequence Deletion, Support, Non-U.S. Gov't, Transcription Factors/metabolism, Transfection, Tumor Cells, Cultured
dx.doi.org/10.1016/0167-4781(94)90059-0, hdl.handle.net/1765/2488
BBA - Gene Structure and Expression
Erasmus MC: University Medical Center Rotterdam

Pruzina, S, Antoniou, M, Hurst, J, Grosveld, F.G, & Philipsen, J.N.J. (1994). Transcriptional Activation by hypersensitive site three of the human β-globin Locus Control Region in murine erythroleukemia cells. BBA - Gene Structure and Expression, 1219(2), 351–360. doi:10.1016/0167-4781(94)90059-0