Stephan W. Glasser

Regulation of surfactant protein gene transcription.

Surfactant protein concentrations are precisely maintained during fetal development and postnatally controlled, at least in part, by the regulation of gene transcription and/or mRNA stability. Together, these mechanisms contribute to the unique temporal-spatial distribution of surfactant protein synthesis that is characteristic of the mammalian lung. Surfactant proteins A, B and C are expressed primarily in subsets of respiratory epithelial cells, wherein their expression is modified by developmental, physiological, humoral and inflammatory stimuli. Cell specific and humoral regulation of surfactant protein transcription is determined by the interactions of a number of nuclear transcription proteins that function in combination, by binding to cis-acting elements located in the 5 regulatory regions of each of the surfactant protein genes. The unique combination of distinct and shared cis-acting elements and transcriptional proteins serves to modulate surfactant protein synthesis in the lung. The present review will summarize efforts to identify the mechanisms contributing to the regulation of surfactant protein gene transcription in the lung, focusing to the nuclear transcription factor, TTF-1 (or thyroid transcription factor-1), a member of the Nkchi2 family of nuclear transcription proteins. A complete review of regulatory aspects of surfactant homeostasis is beyond the scope of the present summary.

NUCLEAR FACTOR I FAMILY MEMBERS REGULATE THE TRANSCRIPTION OF SURFACTANT PROTEIN-C

Transcription of the surfactant protein-C (SP-C) gene is restricted to Type II epithelial cells in the adult lung. We have shown previously that the 0.32-kilobase pair (kb) mouse SP-C promoter is functional in transient transfection assays of the lung epithelial cell-derived cell line, MLE-15, and that thyroid transcription factor 1 (TTF-1) transactivates promoter activity. The 0.32-kb SP-C promoter can be separated into a proximal promoter region (−230 to +18) and an enhancer region (−318 to −230). Three DNase I footprints were mapped in the promoter region (C1 through C3) and two in the enhancer region (C4 and C5). We now show that nuclear factor I (NFI) family members bind to both individual NFI half-sites in footprints C1, C3, and C5, and to a composite site in footprint C4 by competition gel retardation and antibody supershift analyses. Mutational analysis of the 0.32-kb mouse SP-C promoter and transient transfection of MLE-15 cells demonstrated that the NFI binding sites are required for promoter activity in this cell type. Site-specific mutation of the proximal or distal NFI sites drastically reduced transactivation by a co-transfected NFI-A expression vector in HeLa cells. These data indicate that NFI family member(s), binding to sites in both the promoter and enhancer regions, regulate SP-C gene expression in a process independent of TTF-1.

Chromosomal localization of three pulmonary surfactant protein genes in the mouse

Pulmonary surfactant, a protein-phospholipid mixture, maintains surface tension at the lung epithelium/air interface preventing alveolar collapse during respiration. For mammals appropriate developmental production of surfactant is necessary for adaptation to the air breathing environment. Deficiency of pulmonary surfactant results in respiratory distress syndrome (RDS), a leading cause of death in premature infants. Recently, three lung-specific pulmonary surfactant proteins designated SP-A, SP-B, and SP-C have been described. Cloned sequences for the genes that encode each of these proteins have been partially characterized in humans and other species. Analysis of interspecific backcross mice has allowed us to map the chromosomal locations of these three genes in the mouse. The gene encoding SP-A (Sftp-1) and the gene encoding SP-C (Sftp-2) both map to mouse chromosome 14, although at separate locations, while the gene encoding SP-B (Sftp-3) maps to chromosome 6. The mouse map locations determined in this study for the Sftp genes are consistent with the locations of these genes on the human genetic map and the syntenic relationships between the human and the mouse genomes.