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J Biol Chem, Vol. 273, Issue 28, 17917-17925, July 10, 1998
From the Departments of The gut-enriched Krüppel-like factor (GKLF)
is a newly identified zinc finger-containing transcription factor.
Recent studies indicate that GKLF binds to a core DNA sequence of
5'-(G/A)(G/A)GG(C/T)G(C/T)-3', which is found in an endogenous
cis element, the basic transcription element (BTE) of the
cytochrome P-450IA1 (CYP1A1) promoter. The present study
characterizes the ability of GKLF to regulate CYP1A1 expression. By electrophoretic mobility gel shift assay (EMSA) and
methylation interference assay, GKLF was found to bind BTE in a manner
similar to several other transcription factors known to interact with
BTE including Sp1 and BTEB. Cotransfection studies in Chinese hamster
ovary cells showed that GKLF inhibited the CYP1A1 promoter
in a dose- and BTE-dependent manner. The same experiments
also revealed that BTE was responsible for a significant portion of the
CYP1A1 promoter activity. EMSA of nuclear extracts from
Chinese hamster ovary cells showed that Sp1 and Sp3 were two major
proteins that interacted with BTE. Additional cotransfection studies
showed that GKLF inhibited Sp1-mediated activation of the
CYP1A1 promoter. In contrast, GKLF enhanced
Sp3-dependent suppression of the same promoter. Moreover,
the ability of GKLF to inhibit Sp1-dependent
transactivation was in part due to physical interaction of the two
proteins. These findings indicate that GKLF is a negative regulator of
the CYP1A1 promoter in a BTE-dependent fashion
and that this inhibitory effect is in part mediated by physical
interaction with Sp1.
The Gut-enriched Krüppel-like Factor Suppresses the
Activity of the CYP1A1 Promoter in an
Sp1-dependent Fashion
,,
Medicine and
Biological Chemistry, The Johns Hopkins University School of
Medicine, Baltimore, Maryland 21205 and the ¶ Department of
Chemistry, Tohoku University, Sendai 980, Japan
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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