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J. Biol. Chem., Vol. 275, Issue 40, 31051-31060, October 6, 2000
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From the Departments of Surfactant protein D (SP-D) plays roles in
pulmonary host defense and surfactant homeostasis and is increased
following lung injury. Because AP-1 proteins regulate cellular
responses to diverse environmental stimuli, we hypothesized that the
conserved AP-1 motif (at
Proximal Promoter of the Surfactant Protein D Gene
REGULATORY ROLES OF AP-1, FORKHEAD BOX, AND GT BOX BINDING
PROTEINS*
,§,,, and Pathology and Immunology and
¶ Internal Medicine, Washington University School of Medicine,
St. Louis, Missouri 63110
109) and flanking sequences in the human
SP-D promoter contribute to the regulation of SP-D expression. The AP-1
sequence specifically bound to fra-1, junD, and
junB in H441 lung adenocarcinoma nuclear extracts.
Mutagenesis of the AP-1 motif in a chloramphenicol acetyltransferase reporter construct containing 285 base
pairs of upstream sequence nearly abolished promoter activity,
and co-transfection of junD significantly increased wild
type but not mutant promoter activity. The sequence immediately
downstream of the AP-1 element contained a binding site for HNF-3
(FOXA), and simultaneous mutation of this site
(fox-d) and an upstream FoxA binding site (277, fox-u) caused a 4-fold reduction in chloramphenicol
acetyltransferase activity. Immediately upstream of the AP-1-binding
site, we identified a GT box-containing positive regulatory element.
Despite finding regions of limited homology to the thyroid
transcription factor 1-binding site, SP-D promoter activity did not
require thyroid transcription factor 1. Thus, transcriptional
regulation of SP-D gene expression involves complex interactions with
ubiquitous and lineage-dependent factors consistent with
more generalized roles in innate immunity.
*
This work was supported by National Institutes of Health
Grants HL-29594, HL-44015, and HL-56244.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 © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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