Nucleic Acids Research, 1992, Vol. 20, No. 2 237-243
© 1992
MOLECULAR BIOLOGY |
The LCR-like 
-globin positive regulatory element functions as an enhancer in transiently transfected cells during erythroid differentiation
Sir William Dunn School of Pathology, University of Oxford Oxford OX1 3RE, UK
Received October 25, 1991. Accepted December 18, 1991.
A positive regulatory element (PRE) similar to the locus control region (LCR) of the human ß-gIobin gene cluster has recently been identified 40 kb upstream of the human 
-globin mRNA cap site (Higgs D.R. W.G. Wood, A.P. Jarman, J.Sharpe, J. Lida, I.M. Pretorius, and H. Ayyub. 1990). We investigated the influence of the 
PRE on human -globin promoter activity in transiently transfected cells. The introduction of the PRE into -globin promoter/CAT expression constructs increased -globin promoter activity by 15–30 fold in a human erythroid cell line (Putko) as well as in mouse erythroleukemia cells (MELCs) induced with hexamethylene bisacetamide (HMBA). When these constructs were introduced into uninduced MELCs or HeLa cells, only a 2–3 fold increase in -globin promoter activity was observed. Deletion of 600 bp of -globin 5' flanking sequences containing six putative SP1-binding sites had no significant effect on levels of -globin promoter enhancement by the PRE. We further demonstrated that the PRE and HS2 of the ß-LCR could similarly enhance transcriptional activity of the SV40 early promoter in HMBA induced MELCs. Finally, we showed that -globin promoter activity in the presence of the PRE increased with continued HMBA exposure and was coincident with transcriptional activation of endogenous globin genes.
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