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J Biol Chem, Vol. 273, Issue 5, 3060-3067, January 30, 1998
From the Gastrointestinal Unit and Center for the Study of
Inflammatory Bowel Disease, Department of Medicine, Massachusetts
General Hospital and Harvard Medical School,
Boston, Massachusetts 02114
Intestinal trefoil factor (ITF) is selectively
expressed in goblet cells of the small and large intestinal mucosa.
Detailed analysis of the rat ITF (RITF) promoter was undertaken by
transient transfection and gel mobility shift assays (GMSAs) using the
goblet cell-like LS174T colon cancer-derived cell line. Various lengths of wild-type or mutant constructs of the 5
Identification of a Goblet Cell-specific Enhancer Element in the
Rat Intestinal Trefoil Factor Gene Promoter Bound by a Goblet Cell
Nuclear Protein
-flanking region were linked
to the pXP2 reporter gene luciferase. Expression of 
118 RITF was
significantly decreased compared with 154 RITF, and transfection with
an 18-base pair construct (141 to 124) resulted in more than 5-fold
greater expression than transfection with the promoterless pXP2 gene
construct alone. Using various synthetic oligonucleotide mutants, GMSAs
revealed that only a 9-base pair sequence (CCCCTCCCC) in this element
was required for specific binding, overlapping but distinct from a
Sp1-like element. GMSA demonstrated that this element was specifically
bound by nuclear proteins from intestinal cells with a goblet cell-like
phenotype. These studies demonstrate that a 9-base pair element
(goblet cell response element) between 154 and 118 in the RITF
promoter gene is a cis-active element bound by a distinct nuclear
transcription factor and is capable of directing intestine and goblet
cell-specific expression.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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