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Mol. Cell. Biol., 02 1995, 852-860, Vol 15, No. 2
JJ Bieker and CM Southwood
Erythroid Kruppel-like factor (EKLF) is an erythroid cell-specific DNA-
binding protein that activates transcription from the beta-globin CACCC
element, a functionally important and evolutionarily conserved component of
globin as well as other erythroid cell-specific promoters and enhancers. We
have attempted to elucidate the molecular role of EKLF in
erythrocyte-specific transcriptional activation. First, in vivo and in
vitro analyses have been used to demonstrate that the level of activation
by EKLF is dependent on the orientation and number of CACCC elements, that
EKLF contains separable activation and DNA-binding domains, and that the
EKLF proline-rich region is a potent activator in CV-1 cells when fused to
a nonrelated DNA-binding module. Second, we have established a transient
assay in murine erythroleukemia cells in which reproducible levels of a
reporter can be induced when linked to a locus control region
enhancer-beta-globin promoter and in which induction is abolished when the
promoter CAC site is mutated to a GAL site. Third, we demonstrate that the
EKLF transactivation region, when fused to the GAL DNA-binding domain, can
restore inducibility to this mutated construct and that this inducibility
exhibits activator-, promoter-, and cell-type specificity. These results
demonstrate that EKLF provides a crucial transactivation function for
globin expression and further reinforce the idea that EKLF is an important
regulator of CACCC element-directed transcription in erythroid cells.
Copyright © 1995, American Society for Microbiology
The erythroid Kruppel-like factor transactivation domain is a critical component for cell-specific inducibility of a beta-globin promoter
Brookdale Center for Molecular Biology, Mount Sinai School of Medicine, New York, New York 10029.
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