Molecular cloning and characterization of Japanese eel ovarian P450c17 (CYP17) cDNA.
As a first step in investigating the mechanism underlying the steroidogenic shift from the production of ovarian androgens (vitellogenic stage) to that of 17alpha-hydroxylated progestins (maturational stage) in Japanese eel during induced oogenesis, a cDNA encoding Japanese eel (Anguilla japonica) ovarian P450c17 (CYP17: steroid 17alpha-hydroxylase/C(17-20) lyase) was cloned and sequenced. This cDNA contained the complete coding region representing 510 amino acid residues, which showed high sequence homology to those of rainbow trout (74%) and mammals (45-55%). The protein encoded by this cDNA possessed high enzymatic activities of 17alpha-hydroxylase and C(17-20) lyase, thus quickly converting pregnenolone and progesterone to their respective delta(4) and delta(5) C19 products. P450c17 produced a single transcript of 2.4 kb in length, as assessed by Northern blot. Transcript levels of this enzyme significantly increased throughout artificially induced ovarian development. Considering this together with the previous data showing that C(17-20) lyase activity decreased from the vitellogenic to the maturational stage, whereas 17alpha-hydroxylase activity increased, the present data suggest that changes in C(17-20) lyase activity (the production of androgens) do not depend on transcriptional changes of the P450c17 gene.
Kazeto
Y
Y
Department of Biology, Faculty of Fisheries, Hokkaido University, 3-1-1 Minato-cho, Hakodate, Hokkaido, 041-8611, Japan. fire@fish.hokudai.ac.jp
Ijiri
S
S
Todo
T
T
Adachi
S
S
Yamauchi
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D019556:COS Cells; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D004524:Eels / Q000502:physiology*; D005260:Female; D015723:Gene Library; D008969:Molecular Sequence Data; D010053:Ovary / Q000201:enzymology*; D011061:Poly A / Q000096:biosynthesis / Q000235:genetics; D011284:Pregnenolone / Q000096:biosynthesis; D011374:Progesterone / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013045:Species Specificity; D013254:Steroid 17-alpha-Hydroxylase / Q000096:biosynthesis* / Q000235:genetics
D018076:DNA, Complementary; D012333:RNA, Messenger; D011061:Poly A; D011374:Progesterone; D011284:Pregnenolone; D013254:Steroid 17-alpha-Hydroxylase