Molecular cloning and characterization of the rat ovarian 20 alpha-hydroxysteroid dehydrogenase gene.
The rat 20 alpha-hydroxysteroid dehydrogenase (20 alpha-HSD) is an enzyme responsible for the catabolism of progesterone to the inactive 20 alpha-hydroxprogesterone. We have previously shown that the expression of this enzyme is not regulated by post-translational modification, but at the level of transcription. In this study we have established that the 20 alpha-HSD gene contains nine exons and have isolated a 2.5 kb promoter region. The transcription start site was identified and a TATA box was found. 5' deletions of this promoter significantly decreased basal promoter activity. Treatment with forskolin led to a dose dependent inhibition of the 2.5kg-20 alpha-HSD-luciferase construct. Computer analysis identified one CRE, two Nur77 response elements, two putative AP1 sites and one progesterone response element half-site. In summary, we have identified and partially characterized the promoter region of the rat ovarian 20 alpha-HSD and demonstrated that the regulatory elements for 20 alpha-HSD are present within a 2.5 kb 5' flanking region of the gene.
Zhong
L
L
Department of Physiology and Biophysics, University of Illinois at Chicago 60612, USA.
Ou
J
J
Barkai
U
U
Mao
J F
JF
Frasor
J
J
Gibori
G
G
D015089:20-Hydroxysteroid Dehydrogenases / Q000235:genetics*; D043607:20-alpha-Hydroxysteroid Dehydrogenase; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D005576:Colforsin / Q000494:pharmacology; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D005091:Exons; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D007438:Introns; D008969:Molecular Sequence Data; D010053:Ovary / Q000201:enzymology*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D051381:Rats; D017384:Sequence Deletion; D016385:TATA Box; D014162:Transfection
D017931:DNA Primers; D018076:DNA, Complementary; D005576:Colforsin; D015089:20-Hydroxysteroid Dehydrogenases; D043607:20-alpha-Hydroxysteroid Dehydrogenase