European sea bass ( Dicentrarchus labrax L.) cytochrome P450arom: cDNA cloning, expression and genomic organization

Abstract

Cytochrome P450arom, a key enzyme in the hormonal steroidogenic pathway, mediates the conversion of androgens to estrogens. This work describes the molecular cloning of the cDNA encoding the European sea bass ( Dicentrarchus labrax L.) cytochrome P450arom by means of reverse transcriptase and polymerase chain reaction (RT–PCR) and 5′ and 3′-rapid amplification of cDNA ends (RACE) analyses. The cDNA is 1822 bp in length and encodes a putative protein of 517 amino acids. Northern blot analysis revealed that the ovary expressed a transcript of about 2.2 kb in size. Analysis of the deduced amino acid sequence indicated 62–86% identity with ovarian P450arom of other teleost fish, the highest identity being found with the Japanese flounder, Paralichthys olivaceous. Identity was lower (56–65%) with the P450arom forms first reported in teleost brain. Only 52% identity was observed with the corresponding fragment of the cartilaginous fish, Dasyatis sabina. RT–PCR revealed that the sea bass P450arom mRNA was also expressed, at low levels, in testis and brain. Between the 5′ and 3′-untranslated terminal regions (UTR), the sea bass CYP19 gene contains eight introns. All introns conform to the GT/AG rule for RNA splicing and are inserted in exactly the same positions as those found in Oryzias latipes and the human CYP19 gene.