Document Type

Article

Publication Date

7-1-1997

Publication Title

Nucleic Acids Research

Department

Geisel School of Medicine

Abstract

HS-40 is the major regulatory element of the human α-globin locus, located 40 kb upstream of the ζ-globin gene. To test for potential interactions between HS-40 and the β- or the γ-globin gene promoters in stable transfection assays, the HS-40 core sequence was cloned upstream of either the β promoter or the γ promoter driving the neomycin phosphotransferase gene and enhancer activity was measured using a colony assay. In K562 or in MEL cells, enhancer activity of HS-40 was higher than that of the individual core sequences of the DNase I hypersensitive sites (HS) of the β-globin locus control region (LCR), and ∼60% of the enhancer activity of a 2.5 kb μLCR, which contains the core elements of DNase I hypersensitive sites 1–4. In contrast to the synergistic interaction between the DNase I hypersensitive sites of β locus LCR, combination of HS-40 with these DNase I hypersensitive sites failed to display cooperativity in K562 cells and inhibited enhancer function in MEL cells. Inhibition of enhancer function was also observed when two copies of the HS-40 were arranged tandemly. We conclude that the core element of HS-40 (i) is a powerful enhancer of γ- and β-globin gene expression, (ii) in contrast to other classical enhancers, acts best as a single copy, (iii) does not cooperate with the regulatory elements of the β-globin locus control region.

DOI

10.1093/nar/25.14.2917

Original Citation

Chen H, Lowrey CH, Stamatoyannopoulos G. Analysis of enhancer function of the HS-40 core sequence of the human alpha-globin cluster. Nucleic Acids Res. 1997;25(14):2917-2922. doi:10.1093/nar/25.14.2917

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