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J. Biol. Chem., Vol. 277, Issue 31, 27659-27667, August 2, 2002
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From the GATA-4, -5, and -6 zinc finger and
hepatocyte nuclear factor-1
Physical Interaction between GATA-5 and Hepatocyte Nuclear
Factor-1
Results in Synergistic Activation of the Human
Lactase-Phlorizin Hydrolase Promoter*
§¶
,,,,,§¶, and§¶§§¶¶
Division of Gastroenterology and Nutrition,
Department of Medicine, Children's Hospital, Boston, Massachusetts
02115, the § Department of Pediatrics, Harvard Medical
School, Boston, Massachusetts 02115, the ¶ Department
of Pediatrics, New England Medical Center, Boston, Massachusetts 02111, the §§ Gerald J. and Dorothy R. Friedman School of
Nutrition Science and Policy, Tufts University, Medford, Massachusetts
02155, the ** Division of Gastroenterology,
Department of Medicine, University of Pennsylvania, Philadelphia,
Pennsylvania 19104, the Department of Medicine, Free
University of Amsterdam, Amsterdam, The Netherlands 1081HV,
and the Department of Medicine, University of
Amsterdam, Amsterdam, The Netherlands 1100DD
(HNF-1) homeodomain transcription
factors are expressed in the intestinal epithelium and synergistically
activate the promoter of intestinal genes. Here, we demonstrate that
GATA-5 and HNF-1 physically associate both in vivo and
in vitro and that this interaction is necessary for
cooperative activation of the lactase-phlorizin hydrolase
promoter. Furthermore, physical association is mediated by the
C-terminal zinc finger of GATA factors and the homeodomain of HNF-1.
Deletion of HNF-1 activation domains or interruption of
HNF-1-binding sites in the lactase-phlorizin hydrolase promoter resulted in a complete loss of cooperativity, whereas deletion of
GATA-5 activation domains or interruption of GATA-binding sites resulted in a reduction, but not an elimination, of cooperativity. We
hypothesize that GATA/HNF-1 cooperativity is mediated by HNF-1 through its activation domains, which are oriented for high levels of
activation through binding to DNA and physical association with GATA
factors. These data suggest a paradigm whereby intestine-specific gene
expression is regulated by unique interactions among tissue-restricted transcription factors coexpressed in the intestine. Parallel mechanisms in other tissues as well as in Drosophila suggest that zinc
finger/homeodomain interactions are an efficient pathway of cooperative
activation of gene transcription that has been conserved throughout evolution.
*
This work was supported by NIDDK Grant R37-DK-32658
and Silvio O. Conte Digestive Disease Core Center Grant P30-DK-34928
from the National Institutes of Health, by a grant from the Nutricia Research Foundation (to H. M. v. W.), and by an interim support grant from the New England Medical Center (to S. D. K.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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