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Molecular and Cellular Biology, December 2003, p. 9014-9024, Vol. 23, No. 24
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.24.9014-9024.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Nuclear Factor I/Thyroid Transcription Factor 1 Interactions Modulate Surfactant Protein C Transcription

Cindy J. Bachurski,1* Guan Hu Yang,1 Tracey A. Currier,1 Richard M. Gronostajski,2 and Dihua Hong1

Division of Pulmonary Biology, Cincinnati Children's Research Foundation, Cincinnati, Ohio 45229,1 Department of Biochemistry, State University of New York at Buffalo, Buffalo, New York 142142

Received 19 May 2003/ Returned for modification 21 July 2003/ Accepted 18 September 2003

Surfactant protein C (SP-C; Sftpc) gene expression is restricted to pulmonary type II epithelial cells. The proximal SP-C promoter region contains critical binding sites for nuclear factor I (NFI) and thyroid transcription factor 1 (TTF-1; also called Nkx2.1). To test the hypothesis that NFI isoforms interact with TTF-1 to differentially regulate SP-C transcription, we performed transient transfection assays in JEG-3 cells, a choriocarcinoma cell line with negligible endogenous NFI or TTF-1 activity. Cotransfection of NFI family members with TTF-1 induced synergistic activation of the SP-C promoter that was further enhanced by p300. TTF-1 directly interacts with the conserved DNA binding and dimerization domain of all NFI family members in coimmunoprecipitation and mammalian two-hybrid experiments. To determine whether SP-C expression is regulated by NFI in vivo, a chimeric fusion protein containing the DNA binding and dimerization domain of NFI-A and the Drosophila engrailed transcriptional repression domain (NFIen) was conditionally expressed in mice under control of a doxycycline-inducible transgene. Induction of NFIen in a subset of type II cells inhibited SP-C gene expression without affecting expression of TTF-1 in doxycycline-treated double-transgenic mice. Taken together, these findings support the hypothesis that NFI family members interact with TTF-1 to regulate type II cell function.

* Corresponding author. Mailing address: Division of Pulmonary Biology, Cincinnati Children's Research Foundation, 3333 Burnet Ave., Cincinnati, OH 45229-3039. Phone: (513) 636-8120. Fax: (513) 636-7868. E-mail: Cindy.Bachurski{at}cchmc.org.

Molecular and Cellular Biology, December 2003, p. 9014-9024, Vol. 23, No. 24
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.24.9014-9024.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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