Characterization of the DNase I Hypersensitive Site 3 of the Human β Globin Gene Domain

• Published in: Blood Vol. 81; no. 10; pp. 2781 - 2790
• Main Authors: Fleenor, Donald E., Kaufman, Russel E.
• Format: Journal Article
• Language: English
• Published: Washington, DC Elsevier Inc 15-05-1993; The Americain Society of Hematology
• Subjects: Analytical, structural and metabolic biochemistry; Base Sequence; Binding Sites; Biological and medical sciences; Cell Nucleus - metabolism; Cloning, Molecular; Deoxyribonuclease I - metabolism; DNA, Neoplasm - genetics; DNA, Neoplasm - isolation & purification; DNA, Neoplasm - metabolism; Fundamental and applied biological sciences. Psychology; Gene Library; Genes, Regulator; Globins - genetics; Hemoproteins; Humans; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Metalloproteins; Molecular Sequence Data; Multigene Family; Oligodeoxyribonucleotides; Proteins; Restriction Mapping; Substrate Specificity; Tumor Cells, Cultured; Characterization; Globin; Human; Deoxyribonuclease I; Enzymatic activity; Enzyme; Beta-Peptide chain; Exploration
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.V81.10.2781.2781

The members of the human β globin gene family are flanked by strong DNase I hypersensitive sites. The collection of sites 5′ to the ϵ globin gene is able to confer high levels of expression of linked globin genes, but a function has not been assigned to the site 3′ to the β globin gene (3′HS1). Our analysis of this DNase I super hypersensitive site shows that the region is composed of multiple DNase I sites. By examination of the DNA sequence, we have determined that the region is very A/T-rich and contains topoisomerase II recognition sequences, as well as several consensus binding motifs for GATA-1 and AP-1/NF-E2. Gel mobility shift assays indicate that the region can interact in vitro with GATA-1 and AP-1/NF-E2, and functional studies show that the region serves as a scaffold attachment region in both erythroid and nonerythroid cell lines. Whereas many of the physical features of 3′HS1 are shared by 5′HS2 (a component of the 5′ locus control region), transient expression studies show that 3′ HS1 does not share the erythroid-specific enhancer activity exhibited by 5′HS2.