Nucleic Acids Research, Vol 27, Issue 23 4562-4569, Copyright © 1999 by Oxford University Press
CS Mahatan, KH Kaestner, DE Geiman and VW Yang
Gut-enriched Kruppel-like factor (GKLF, KLF4) is an epithelial-specific
transcription factor whose expression is associated with growth arrest. In
order to understand the mechanisms regulating expression of the gene
encoding GKLF, we isolated a genomic clone containing murine GKLF. The gene
spans 5.3 kb and contains four exons. A major start site of transcription
was mapped to an adenine residue 601 nt 5' of the translation initiation
codon. An additional 1 kb of the 5'-flanking region was sequenced and found
to contain multiple cis -elements homologous to the binding sites of
several established transcription factors including Sp1, AP-1, Cdx, GATA,
and USF. In particular, three closely spaced GC-boxes 5' of the TATA box
resemble the established binding site for GKLF. DNase I protection and
electrophoretic mobility shift assays verified that recombinant GKLF bound
to each of the three GC-boxes. In co-transfection experiments, GKLF
transactivated a reporter gene linked to the GKLF 1 kb 5'-flanking region,
as did Sp1, Sp3 and Cdx-2. Mutations of one or both of the first and second
GC- boxes in the promoter resulted in diminished transactivation by GKLF.
These results demonstrate that the 5'-flanking sequence of the mouse GKLF
gene functions as a promoter and is subject to autoregulation by its own
gene product.
ARTICLES
Characterization of the structure and regulation of the murine gene encoding gut-enriched Kruppel-like factor (Kruppel-like factor 4)
Department of Medicine, The Johns Hopkins School of Medicine, Baltimore, MD 21205, USA.
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