Teleost ovarian carbonyl reductase-like 20beta-hydroxysteroid dehydrogenase: potential role in the production of maturation-inducing hormone during final oocyte maturation.
17alpha,20beta-Dihydroxy-4-pregnen-3-one is the major oocyte maturation-inducing hormone of several teleost species. Gonadotropin-induced increase in ovarian 20beta-hydroxysteroid dehydrogenase activity is essential for the synthesis of maturation-inducing hormone. Cloning and expression studies suggest that ayu (Plecoglossus altivelis) ovarian carbonyl reductase can function as 20beta-hydroxysteroid dehydrogenase. The amino acid sequence deduced from the isolated cDNA had 276 amino acid residues and shared approximately 60% homology with mammalian and teleostean carbonyl reductases. The sequence data search showed that the ayu cDNA clone belongs to the short-chain dehydrogenase/reductase family. The clear lysate prepared from Escherichia coli harboring the cDNA catalyzed the production of maturation-inducing hormone. Its identification was confirmed by two-dimensional, thin-layer chromatography followed by recrystallization. Purification of the E. coli-expressed cDNA product revealed that it possessed both carbonyl reductase and steroid dehydrogenase activities, and 17alpha-hydroxyprogesterone, the endogenous immediate precursor of maturation-inducing hormone, was one of the preferred substrates. Furthermore, Northern blot analysis denoted that the transcripts are present both in fully grown, immature ovarian follicles and at higher levels in mature ovarian follicles. These results demonstrate that the carbonyl reductase of ayu ovary is involved in the production of maturation-inducing hormone, and they provide evidence for a novel physiological role of this enzyme in the final maturation of oocytes. Based on its functional properties, the enzyme can be referred to as carbonyl reductase-like 20beta-hydroxysteroid dehydrogenase.
Tanaka
Minoru
M
Laboratory of Reproductive Biology, National Institute for Basic Biology, Okazaki 444-8585, Japan.
Nakajin
Shizuo
S
Kobayashi
Daisuke
D
Fukada
Sachiko
S
Guan
Guijun
G
Todo
Takashi
T
Senthilkumaran
Balasubramanian
B
Nagahama
Yoshitaka
Y
D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D003349:Cortisone Reductase / Q000235:genetics* / Q000502:physiology*; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000201:enzymology; D005260:Female; D005399:Fishes / Q000502:physiology*; D015723:Gene Library; D006908:Hydroxyprogesterones / Q000378:metabolism*; D008540:Meiosis / Q000502:physiology; D008969:Molecular Sequence Data; D009865:Oocytes / Q000254:growth & development*; D010053:Ovary / Q000201:enzymology*
D018076:DNA, Complementary; D006908:Hydroxyprogesterones; C007433:17,20-dihydroxy-4-pregnen-3-one; D000429:Alcohol Oxidoreductases; D003349:Cortisone Reductase