Molecular cloning of the gene for the human prostaglandin transporter hPGT: gene organization, promoter activity, and chromosomal localization

Abstract

Prostaglandins (PGs) play diverse and important roles in human health and disease. We recently identified the first known PG transporter cDNA in the rat (PGT) and human (hPGT). To aid in the analysis of any possible human disease caused by mutations in PGT, we have cloned and characterized the hPGT gene. The gene exists as a single copy in the human genome and is comprised of 14 exons distributed over approximately 95 kb. Two introns disrupt putative trans-membrane spans of the coding region; each of these sites is near a highly conserved charged residue. The approximately 250 bp immediately 5' to the start of exon 1 contain a TATAAA sequence (TATA box), a transcription initiation (Inr) consensus (CTCANTCT), two Sp 1 sequences (GGGCGG), and a cAMP response element (CGGCGTCA). Ligation of approximately 3.5 kb of 5' flanking sequence to a luciferase reporter yielded > 15-fold activity above background when expressed in A549 human lung epithelial cells. PCR-based monochromosomal somatic cell hybrid mapping and fluorescence in situ hybridization localized hPGT to chromosome 3q21. Three microsatellites were identified, one of which was demonstrated to be polymorphic in unrelated individuals and may be useful in evaluating PGT as a candidate gene in human disease.