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Am J Physiol Lung Cell Mol Physiol 276: L398-L404, 1999;
1040-0605/99 $5.00
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Vol. 276, Issue 3, L398-L404, March 1999

Inhibition of hSP-B promoter in respiratory epithelial cells by a dominant negative retinoic acid receptor

Manely Ghaffari, Jeffrey A. Whitsett, and Cong Yan

Division of Pulmonary Biology, Children's Hospital Medical Center, Cincinnati, Ohio 45229-3039

Retinoic acid (RA) receptors (RARs) belong to the nuclear hormone receptor superfamily and play important roles in lung differentiation, growth, and gene regulation. Surfactant protein (SP) B is a small hydrophobic protein synthesized and secreted by respiratory epithelial cells in the lung. Expression of the SP-B gene is modulated at the transcriptional and posttranscriptional levels. In the present work, immunohistochemical staining revealed that RAR-alpha is present on day 14.5 of gestation in the fetal mouse lung. To assess whether RAR is required for SP-B gene transcription, a dominant negative mutant human (h) RAR-alpha 403 was generated. The hRAR-alpha 403 mutant was transcribed and translated into the truncated protein product by reticulocyte lysate in vitro. The mutant retained DNA binding activity in the presence of retinoid X receptor-gamma to an RA response element in the hSP-B promoter. When transiently transfected into pulmonary adenocarcinoma epithelial cells (H441 cells), the mutant hRAR-alpha 403 was readily detected in the cell nucleus. Cotransfection of the mutant hRAR-alpha 403 repressed activity of the hSP-B promoter and inhibited RA-induced surfactant proprotein B production in H441 cells, supporting the concept that RAR is required for hSP-B gene transcription in vitro.

human surfactant protein B; lung development; nuclear receptors


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