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J Biol Chem, Vol. 273, Issue 23, 14347-14354, June 5, 1998
From the Department of Molecular Genetics, Biochemistry, and
Microbiology, University of Cincinnati,
Cincinnati, Ohio 45267-0524
Erythroid Krüppel-like factor (EKLF) is a
zinc finger transcription factor required for 
-globin gene
expression and is implicated as one of the key factors necessary for
the fetal to adult switch in globin gene expression. In an effort to
identify factors involved in the expression of this important
erythroid-specific regulatory protein, we have isolated the mouse EKLF
gene and systematically analyzed the promoter region. Initially, a
reporter construct with 1150 base pairs of the EKLF 5'-region was
introduced into transgenic mice and shown to direct erythroid-specific
expression. We continued the expression studies in erythroid cells and
have identified a sequence element consisting of two GATA sites
flanking an E box motif. The three sites act in concert to elevate the transcriptional activity of the EKLF promoter. Each site is essential for EKLF expression indicating that the three binding sites do not work
additively, but rather function as a unit. We further show that GATA-1
binds to the two GATA sites and present evidence for binding of another
factor from erythroid cell nuclear extracts to the E box motif. These
results are consistent with the formation of a quaternary complex
composed of an E box dimer and two GATA-1 proteins binding at a
combined GATA-E box-GATA activator element in the distal EKLF
promoter.
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