Characterization of the Gene and Promoter for RTI40, a Differentiation Marker of Type I Alveolar Epithelial Cells

In an effort to understand the processes that establish and maintain the differentiated state of the alveolar epithelium, we have analyzed the gene for rat type I cell 40 kD protein (RTI40), an apical integral plasma membrane protein expressed in type I but not type II alveolar epithelial cells. The RTI40 gene spans 35 kilobase pairs; it contains 6 exons and at least 6 rat Identifier repetitive elements. Three exons encode the predicted RTI40 extracellular domain and one encodes the single transmembrane spanning domain. The final exon encodes one amino acid followed by a stop codon. RTI40 gene transcription starts downstream from a TATA homology, which is immediately adjacent to putative binding sites for thyroid transcription factor 1 and Sp1. In H441 cell transfections, mutagenesis of a 5'-flanking fragment (-2496 to +104) revealed two regions that contribute to promoter activity: -1247 through -795 and -163 through -81. Heterologous promoter fusion experiments suggest that a cooperative interaction between these regions activates transcription. In transfected type II cells, deletion across the proximal region produced a 6-fold drop in promoter activity, whereas deletion across the distal region was without apparent effect. These results provide a foundation to analyze further the factors that govern alveolar epithelial cell phenotype.