RRC ID 1263
Author Kazeto Y, Ijiri S, Matsubara H, Adachi S, Yamauchi K.
Title Molecular cloning and characterization of 3beta-hydroxysteroid dehydrogenase/Delta5-Delta4 isomerase cDNAs from Japanese eel ovary.
Journal J Steroid Biochem Mol Biol
Abstract 3beta-hydroxysteroid dehydrogenase/Delta(5)-Delta(4) isomerase (3beta-HSD) is a crucial steroidogenic enzyme which catalyzes an essential step in the biosynthesis of all classes of steroid hormones. Two closely related cDNAs, encoding Japanese eel ovarian types I and II 3beta-HSD, were cloned and characterized. Both cDNAs putatively encoded 375 amino acid residues sharing high sequence homology with those of rainbow trout (71%) and mammalian (approximately 45-50%) 3beta-HSD. Transient expression of types I and II 3beta-HSD in COS-7 cells revealed that both proteins possess 3beta-hydroxysteroid dehydrogenase as well as Delta(5)-Delta(4) isomerase activity for both pregnenolone and dehydroepiandrosterone, with the preference of pregnenolone over dehydroepiandrosterone as substrate, although the type I protein is more active than the type II. By northern blot analysis, a single band of the 3beta-HSD transcript of approximately 1.5kb in length was observed in ovarian tissue and the total transcript abundance of both 3beta-HSDs remained constant throughout ovarian development artificially induced by gonadotropin-rich salmon pituitary homogenate. This lack of change in 3beta-HSD transcript abundance during ovarian development did not correlate with the fluctuation of its enzymatic activity reported previously, which may suggest that changes in 3beta-HSD activity during ovarian development may be, in part, post-transcriptionally regulated in the Japanese eel ovary.
Volume 85(1)
Pages 49-56
Published 2003-5-1
DOI 10.1016/s0960-0760(03)00138-9
PII S0960076003001389
PMID 12798356
MeSH 17-Hydroxysteroid Dehydrogenases / chemistry 17-Hydroxysteroid Dehydrogenases / genetics* 17-Hydroxysteroid Dehydrogenases / metabolism Amino Acid Sequence Androstenedione / metabolism Anguilla / genetics Anguilla / growth & development Anguilla / metabolism* Animals Base Sequence Blotting, Northern COS Cells Chlorocebus aethiops Cloning, Molecular DNA, Complementary / genetics Female Gene Expression Regulation, Developmental Gene Expression Regulation, Enzymologic Molecular Sequence Data Ovary / enzymology* Ovary / growth & development Progesterone / metabolism RNA / chemistry RNA / genetics Reverse Transcriptase Polymerase Chain Reaction Sequence Alignment
IF 3.813
Times Cited 24
Human and Animal Cells COS-7(RCB0539)