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J Biol Chem, Vol. 275, Issue 1, 56-62, January 7, 2000
From the Division of Pulmonary Biology, Children's Hospital
Medical Center, Cincinnati, Ohio 45229-3039
Surfactant B (SP-B) is a 79-amino acid peptide
critical to postnatal respiratory adaptation. Expression of SP-B by
respiratory epithelial cells is regulated by developmental and hormonal
influences at the level of gene transcription. Previous studies
supported the role of retinoic acids (RA) and their receptors (RARs) in SP-B gene transcription. In the present study, RAR
Retinoic Acid Stimulation of the Human Surfactant Protein B
Promoter Is Thyroid Transcription Factor 1 Site-dependent*
,,
was
detected in mouse alveolar type II epithelial cells where SP-B is
synthesized and processed. Deletion and site-specific mutagenesis
analysis identified clustered retinoic acid-responsive element sites in the 5'-flanking enhancer region of the hSP-B gene that
bound RAR proteins. RAR coactivators ACTR, SRC-1, and
transcriptional intermediary factor 2 (TIF2) stimulated human (h) SP-B
promoter activity in a dose-dependent fashion in pulmonary
adenocarcinoma H441 cells. In addition, an RAR-associated protein,
CREB-binding protein (CBP), potentiated the effects of RAR on hSP-B
promoter activity in H441 cells. Importantly, RA stimulation of the
hSP-B promoter depends on tissue-specific thyroid transcription factor
(TTF-1) DNA-binding sites. TTF-1 protein synergistically stimulated the
hSP-B promoter with RAR, CBP, and nuclear receptor coactivators in
H441 cells. In addition, TTF-1 interacted directly with RAR and TIF2
in the mammalian two-hybrid system. These findings support a model in which RAR/retinoid X receptor, TTF-1, coactivators, and CBP form a
transcription activation complex in the upstream enhancer region of the
hSP-B gene.
*
This work was supported by an American Lung Association
grant (to C. Y.), NHLBI Grant HL-61803 from the National Institutes of
Health (to C. Y.), and Specialized Center of Research Grants HL-56387
(to J. A. W. and C. Y.) and HL38859 (to J. A. W.).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.
The first two authors contributed equally to this work.
§
To whom correspondence should be addressed: Children's Hospital
Medical Center, Division of Pulmonary Biology, TCHRF, 3333 Burnet Ave.,
Cincinnati, OH 45229-3039. Tel.: 513-636-7990; Fax: 513-636-7868;
E-mail: Cong.Yan@chmcc.org.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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