Rabbit surfactant protein B gene: structure and functional characterization of the promoter.
Surfactant protein B (SP-B) is essential for physiological function of pulmonary surfactant. In the present investigation, we isolated rabbit SP-B gene and determined its nucleotide sequence, transcription start site, and exon/intron organization. The coding region of rabbit SP-B gene is comprised of 6.8 kb of sequence and is organized into 10 introns and 11 exons. By deletion analysis we determined that a region of SP-B gene extending from -236 to +39 nucleotides is sufficient for high-level expression of CAT reporter gene in a cell type-specific manner in the pulmonary adenocarcinoma cell line NCI-H441. Deletion of 5'-flanking sequence to -140 and -71 nucleotides significantly reduced SP-B promoter activity, suggesting that the -236 to +39 region contains cis-DNA elements required for cell type-specific expression. The proximal promoter region of SP-B gene contained DNA sequence motifs for binding thyroid transcription factor 1 (TTF-1) (-113 to -97) and hepatocyte nuclear factor 3(HNF-3) (-91 to -81). Although SP-B gene sequence -140 to +39 did not support high level expression of CAT gene in NCI-H441 cells, it was capable of activation by TTF-1 in HeLa cells, suggesting that the -236 to -140 sequence plays an important role in cell type-specific activation of SP-B promoter.
Margana
R K
RK
Department of Molecular Biology, University of Texas Health Science Center at Tyler 75710, USA.
Boggaram
V
V
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002678:Chimera; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004247:DNA / Q000235:genetics; D004742:Enhancer Elements, Genetic; D017353:Gene Deletion; D015870:Gene Expression; D005796:Genes; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D011510:Proteolipids / Q000235:genetics*; D011663:Pulmonary Surfactants / Q000235:genetics*; D011817:Rabbits; D000074482:Thyroid Nuclear Factor 1; D014157:Transcription Factors / Q000494:pharmacology; D015533:Transcriptional Activation
D015335:Molecular Probes; D009687:Nuclear Proteins; D011510:Proteolipids; D011663:Pulmonary Surfactants; D000074482:Thyroid Nuclear Factor 1; D014157:Transcription Factors; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase