Nucleic Acids Research, 1990, Vol. 18, No. 24 7407-7415
© 1990
Articles |
Structure and function of the enhancer 3' to the human A
, globin gene
Clinical Hematology Branch, National Heart, Lung and Blood Institute Bethesda, MD 20892 1Division of Medical Genetics, Research and Education Instftute Harbor-UCLA Medical Center, Torrance, CA 90502, USA
*To whom correspondence should be addressed at National Institutes of Health, 9000 Rockville Pike, Building 10, Room 7C-103, Bethesda, MD 20892, USA
Received June 26, 1990. Revised October 29, 1990. Accepted October 29, 1990.
An enhancer is located immediately 3' to the Agamma globin gene. We have used DNase I footprinting to map the sites of interaction of nuclear proteins with the DNA sequences of this enhancer. Eight footprints were discovered, distributed over 600 base pairs of DNA. Three of these contain a consensus binding site for the erythroid specific factor GATA-I. Each of these GATA-1 sites had an enhancer activity when inserted into a reporter plasmid and tested in human erythroleukemia cells. Other footprints within the enhancer contained consensus binding sequences for the ubiquitous, positive regulatory proteins AP2 and CBP-1. An Sp1-like recognition sequence was also identified. Synthetic oligonucleotides encompassing two of the footprints generated a slowly migrating complex in gel mobility shift assays. The same complex forms on a fragment of the human gamma globin gene promoter extending from –260 to –200. The DNasel footprint of this protein complex with the enhancer overlapped a sequence, AGGAGGA, found within the binding site for a protein that interacts with the chicken beta globin promoter and enhancer, termed the stage selector element. We propose that this complex of proteins may be involved in the human gamma globin promoter-enhancer interaction.
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