Cloning and sequence analysis of the cDNA encoding P-450 aromatase (P450arom) from a rainbow trout (Oncorhynchus mykiss) ovary; relationship between the amount of P450arom mRNA and the production of oestradiol-17 beta in the ovary.
The enzyme aromatase P-450 (P450arom) catalyses the conversion of androgen to oestrogen. A cDNA insert encoding P450arom was isolated from a rainbow trout (Oncorhynchus mykiss) ovary cDNA library. The insert was sequenced and found to contain an open-reading frame predicted to encode a protein of 522 amino acid residues. The deduced polypeptide is 52% homologous with human, mouse and rat P450arom and 53% homologous with that of chicken. The insert was confirmed to encode P450arom by introducing it into COS-1 monkey kidney tumour cells (COS-1 cells) and detecting the conversion of testosterone to oestradiol-17 beta by radioimmunoassay. The N-terminal region of the deduced polypeptide was 19 amino acids longer than that of the other four species, and was found by hydropathy plotting to be very hydrophobic. Northern blot analysis revealed 2.6 kb RNA transcripts which were present in the trout ovary during vitellogenesis and hybridized to the cDNA insert. In preparations from subsequent stages of ovarian development, no RNA transcripts hybridized to the probe. Since the RNA transcripts are present only during the stage of oestradiol-beta production by the ovarian follicles, oestradiol-17 beta production may be regulated, in part, by the amount of P450arom mRNA present.
Tanaka
M
M
Laboratory of Reproductive Biology, National Institute for Basic Biology, Okazaki, Japan.
Telecky
T M
TM
Fukada
S
S
Adachi
S
S
Chen
S
S
Nagahama
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001141:Aromatase / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004958:Estradiol / Q000096:biosynthesis; D005260:Female; D006801:Humans; D008969:Molecular Sequence Data; D010053:Ovary / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D014337:Trout
D012333:RNA, Messenger; D004958:Estradiol; D004247:DNA; D001141:Aromatase