In vitro analysis of rat surfactant protein A gene expression
Previous findings suggest that the rat proximal promoter segment (rPPS) of the SP-A gene is important in the regulation of the lung-specific expression of the gene. In this report, two regions within the rPPS containing thyroid transcription factor-1 (TTF-1) binding sites are identified that form strong lung- and thyroid- specific DNA-protein complexes. These regions bind nuclear polypeptides with similar apparent molecular mass to TTF-1, suggesting that TTF-1 binds to these regions. Two regions within the rPPS that form weaker lung-specific DNA-protein complexes are also identified. The transcription start site is mapped, and a functional analysis shows that the sequences of the 5' flanking region are sufficient to support in vitro transcription but are not sufficient to reproduce tissue-specific expression. Together, these results show that lung- and thyroid-specific, as well as lung-specific, DNA-protein interactions occur within the rPPS but are not sufficient for the lung-specific expression of the rat SP-A gene to be duplicated in vitro.