Investigation on the transcription factors of porcine 3β-hydroxysteroid dehydrogenase and 17β-hydroxysteroid dehydrogenase genes

The enzymes 3β-hydroxysteroid dehydrogenase (3βHSD) and 17β-hydroxysteroid dehydrogenase (17βHSD) regulate the steroid metabolism in mammals. In this study, we aimed to characterize the steroid related transcription factors at the 5′ flanking region of these two genes. A series of 5′ deletions of approximately 1 kb of 5′-flanking region on both genes were fused to a pGL3 basic vector containing firefly luciferase cDNA, and then transfected to human hepatocellular liver carcinoma cell line (HepG2). Luciferase activity assay indicated the region from − 574 to − 617 bp of the 3βHSD1 promoter, and from − 850 to − 868 bp of 17βHSD7 promoter induced the highest luciferase activity. A putative transcription factor, i.e. the proline and acidic amino acid-rich basic leucine zipper (PAR/bZIP) family of 3βHSD1 gene, and three-amino acid loop extension (TALE) homeodomain class of 17βHSD7 were identified respectively by sequence homology. Gel shift assay further confirmed the binding capacity of the putative elements to nuclear extract. Our study gives new insights to the transcriptional regulation of 3βHSD1 and 17βHSD7 and further hints to their involvement in steroid metabolism.

► We studied steroid related transcription factor of porcine 3βHSD1 and 17βHSD7 genes. ► Core regulation regions of both genes were identified by transfection analysis. ► EMSA further confirmed the binding capacity of putative elements to nuclear extract.