The cDNA cloning and transient expression of an ovary-specific 17beta-hydroxysteroid dehydrogenase of chickens.
A cDNA clone, pc17bHSD, was obtained from the chicken ovarian cDNA library by its partial homology to the cDNA sequence of the rat 17beta-hydroxysteroid dehydrogenase (17beta-HSD). The cDNA insert of pc17bHSD is 979bp long and contains an open reading frame (ORF) of 906bp. The deduced amino acid sequence of the ORF shows 48 and 50% overall identity with those of the rat and the human type-1 17beta-HSD, respectively. Five sequence regions common to the short-chain alcohol dehydrogenase superfamily are well conserved, including the YxxxK sequence motif at the active site. Northern blot hybridization detected a transcript of about 1kb only in ovaries of both sexually immature and mature female chickens. The 17beta-HSD activity, which was highly specific to the interconversion between estrone and estradiol-17beta, was detected in the cytoplasmic fraction of human 293 cells transfected transiently with an expression vector carrying the c17bHSD cDNA sequence, pcDNAI/c17bHSD. From these results, it is concluded that the pc17bHSD is the cDNA clone for the ovary-specific molecular species of 17beta-HSD in chickens.
Wajima
Y
Y
Laboratory of Molecular Biology, Department of Applied Biological Chemistry, Faculty of Agriculture, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai 981-8555, Japan.
Furusawa
T
T
Kawauchi
S
S
Wakabayashi
N
N
Nakabayashi
O
O
Nishimori
K
K
Mizuno
S
S
D015067:17-Hydroxysteroid Dehydrogenases / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002645:Chickens; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004958:Estradiol / Q000378:metabolism; D004970:Estrone / Q000378:metabolism; D005260:Female; D006801:Humans; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010053:Ovary / Q000201:enzymology*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D018076:DNA, Complementary; D012333:RNA, Messenger; D004970:Estrone; D004958:Estradiol; D015067:17-Hydroxysteroid Dehydrogenases; C044256:3 (or 17)-beta-hydroxysteroid dehydrogenase