Gene structure, chromosomal localization and analysis of 3-ketosteroid reductase activity of the human 3(alpha-->beta)-hydroxysteroid epimerase.
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Huang XF, Luu-The V
Gene structure, chromosomal localization and analysis of 3-ketosteroid reductase activity of the human 3(alpha-->beta)-hydroxysteroid epimerase.
Biochim Biophys Acta. 2001 Aug 30;1520(2):124-30.
- PubMed ID
- 11513953 [ View in PubMed]
- Abstract
Following our previous characterization of the first human 3(alpha-->beta)hydroxysteroid epimerase (hHSE), we determined the genomic structure and chromosomal localization of the hHSE gene using fluorescent in situ hybridization (FISH) in this study. The gene spans 23 kb and contains five exons and four introns. FISH mapping assigned this gene to chromosome band 12q13. Primer extension analysis allowed the identification of a single transcription start site at 179 bp upstream from the ATG start codon. The 5'-flanking sequence lacks a typical TATA box in the proximal region of the transcription start site. However, analysis of the 2 kb promoter region revealed the presence of multiple potential transcription factor binding sites. Furthermore, we studied the 3-ketosteroid reductase activity demonstrated by hHSE in intact cells stably expressing the enzyme. It has been known that, in vitro, 3beta-hydroxysteroid dehydrogenase (3beta-HSD) shows both oxidative and reductive activity. Our results showed that hHSE catalyzes the reduction of 3-ketosteroids to form 3beta-hydroxysteroids while 3beta-HSD cannot catalyze this reaction in intact cells. However, hHSE showed 3-keto reductase activity in both microsomal fractions and intact cells. Since intact cells constitute a system which closely reflects in vivo intracellular conditions, we propose that hHSE might contribute to the cellular 3-ketosteroid reductase activity in the peripheral tissues.