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Volume 272, Number 33, Issue of August 15, 1997 pp. 20764-20773
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Phorbol Ester Down-regulation of Lung Surfactant Protein B Gene Expression by Cytoplasmic Trapping of Thyroid Transcription Factor-1 and Hepatocyte Nuclear Factor 3

(Received for publication, April 18, 1997)

A. Suresh Kumar ^¶ , Vugranam C. Venkatesh , Benjamin C. Planer ^¶ , Sheldon I. Feinstein and Philip L. Ballard ^¶

From the  University of Pennsylvania School of Medicine, Institute for Environmental Medicine, Philadelphia, Pennsylvania 19104-6068, the  Department of Pediatrics, University of British Columbia, Vancouver, British Columbia, Canada V5Z 4H4, and the ^¶ Department of Pediatrics, University of Pennsylvania School of Medicine, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania 19104-4318

The lung-specific surfactant protein B (SP-B) is essential for surfactant function and normal respiration. We investigated the role of thyroid transcription factor-1 (TTF-1) and hepatocyte nuclear factor 3 (HNF3) in the down-regulation of SP-B gene expression by phorbol ester in pulmonary adenocarcinoma H441 cells. Responsiveness to 12-O-tetradecanoylphorbol-13-acetate (TPA) localized to the SP-B proximal promoter (140/65 bp) and specifically to binding sites for TTF-1 and HNF3, which act as cell-specific enhancers of SP-B expression. Treatment of cells with TPA (10 nM) caused a time-dependent decrease in both TTF-1 and HNF3 in nuclear extracts and accumulation of both factors in the cytoplasm as assessed by electromobility shift, Western, Southwestern, and immunofluorescence assays. Treatment did not alter the mRNA content or DNA binding activity for either transcription factor. We conclude that down-regulation of SP-B gene expression by phorbol ester involves cytoplasmic trapping and loss of TTF-1 and HNF3 from the nucleus. This mechanism of action is independent of AP-1 and other transcription factors known to be influenced by phorbol ester.

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