Nucleic Acids Research, Vol 26, Issue 3 796-802, Copyright © 1998 by Oxford University Press
JM Shields and VW Yang
The gut-enriched Kruppel-like factor (GKLF) is a recently identified
eukaryotic transcription factor that contains three C2H2zinc fingers. The
amino acid sequence of the zinc finger portion of GKLF is closely related
to several Kruppel proteins, including the lung Kruppel-like factor (LKLF),
the erythroid Kruppel-like factor (EKLF) and the basic transcription
element binding protein 2 (BTEB2). The DNA sequence to which GKLF binds has
not been definitively established. In the present study we determined the
DNA binding sequence of GKLF using highly purified recombinant GKLF in a
target detection assay of an oligonucleotide library consisting of random
sequences. Upon repeated rounds of selection and subsequent
characterization of the selected sequences by base-specific mutagenesis a
DNA with the sequence 5'- G/AG/AGGC/TGC/T-3' was found to contain the
minimal essential binding site for GKLF. This sequence is present in the
promoters of two previously characterized genes: the CACCC element of the
beta-globin gene, which interacts with EKLF, and the basic transcription
element (BTE) of the CYP1A1 gene, which interacts with Sp1 and several
Sp1-like transcription factors. Moreover, the selected GKLF binding
sequence was capable of mediating transactivation of a linked reporter gene
by GKLF in co-transfection experiments. Our results establish GKLF as a
sequence-specific transcription factor likely involved in regulation of
expression of endogenous genes.
ARTICLES
Identification of the DNA sequence that interacts with the gut-enriched Kruppel-like factor
Department of Medicine and Department of Biological Chemistry, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
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