Novel 3beta-hydroxysteroid dehydrogenases from gonads of the Nile tilapia: phylogenetic significance and expression during reproductive cycle

Multiple forms of 3beta-hydroxysteroid dehydrogenase/isomerase (3beta-HSD) and their differential tissue expression pattern have not been shown in any lower vertebrates. In the present study, we report cloning of two novel 3beta-HSDs and two variants from gonads of the Nile tilapia. 3beta-HSD cDNAs...

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Bibliographic Details
Published in:Molecular and cellular endocrinology 2009-02, Vol.299 (2), p.146-152
Main Authors: Senthilkumaran, B, Sudhakumari, C C, Wang, D S, Sreenivasulu, G, Kobayashi, T, Kobayashi, H-K, Yoshikuni, M, Nagahama, Y
Format: Article
Language:English
Subjects:
17-Hydroxysteroid Dehydrogenases - chemistry
17-Hydroxysteroid Dehydrogenases - genetics
17-Hydroxysteroid Dehydrogenases - metabolism
Amino Acid Sequence
Animals
Blotting, Northern
Cercopithecus aethiops
Cichlids - metabolism
Cloning, Molecular
COS Cells
Female
Gene Expression Profiling
Isoenzymes - chemistry
Isoenzymes - genetics
Isoenzymes - metabolism
Male
Molecular Sequence Data
Ovary - enzymology
Phylogeny
Reproduction
Reverse Transcriptase Polymerase Chain Reaction
Testis - enzymology
Transfection
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Summary:Multiple forms of 3beta-hydroxysteroid dehydrogenase/isomerase (3beta-HSD) and their differential tissue expression pattern have not been shown in any lower vertebrates. In the present study, we report cloning of two novel 3beta-HSDs and two variants from gonads of the Nile tilapia. 3beta-HSD cDNAs encode two peptides of 375 (3beta-HSD type-I/variant 1) and 367 (3beta-HSD type-II/variant 1) amino acid residues that share 31.9% homology. 3beta-HSD type-I/variant 1 shared high homology with other piscine counterparts while 3beta-HSD type-II/variant 1 exhibited homology to mammalian DeltaC27-3beta-HSD and multifunctional viral 3beta-HSD. The latter seems to be ancient form among vertebrates. Transiently transfected 3beta-HSDs' open reading frames in COS-7 cells converted exogenous pregnenolone/androsta-5-ene-3beta-17beta-diol to progesterone/testosterone. Tissue distribution pattern of 3beta-HSDs by RT-PCR revealed varied expression pattern. Northern blot analysis of 3beta-HSDs demonstrated steady or gradual rise in transcripts level at different gonadal stages. These data revealed the importance of novel 3beta-HSDs in teleosts and also provided phylogenetic significance.
ISSN:0303-7207
DOI:10.1016/j.mce.2008.11.008