Isolation of cDNA of novel cytochrome P450 1B gene, CYP1B2, from Carp (Cyprinus carpio) and its induced expression in gills.
Cytochrome P450 (CYP) enzymes constitute a multigene family of many endogenous and xenobiotic substances. The CYP1 family is of particular interest in environmental toxicology because its members are dominant in the metabolism of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and aryl amines. A new complementary DNA of the CYP1B subfamily encoding CYP1B2 was isolated from carp liver after intraperitoneal injection with 3-methylcholanthrene (3-MC). The obtained full-length cDNA contained a 5'noncoding region of 161 bp, an open reading frame of 1593 bp coding for 530 amino acids and a stop codon, and a 3'noncoding region of 1457 bp. The predicted molecular weight of the protein was approximately 60.2 kDa. The deduced amino acid sequence of this cDNA was 91% similar to that of our previously reported carp CYP1B1; its similarities with those of the reported CYP1B1s of teleosts and mammals were 60.8, 54.4, 50.8, and 51.4% for plaice, human, rat, and mouse, respectively. The phylogenetic tree of fish and mammalian CYP1 sequences constructed by the protein maximum-likelihood method suggested a relatively recent divergence of CYP1B2 from CYP1B1 in the ancestor of carp and closely related species. Despite the structural similarity of CYP1B2 with CYP1B1, which showed induced expression in 3-MC-treated liver, intestine, and gills with marked constitutive expression in gills, CYP1B2 revealed induced expression in gills but not in liver or intestine, and no detectable constitutive expression in the tissues studied.
El-kady
Mohamed A H
MA
Laboratory of Marine Biotechnology, Faculty of Fisheries, Kagoshima University, 4-50-20 Shimoarata, Kagoshima 890-0056, Japan.
Mitsuo
Ryoichi
R
Kaminishi
Yoshio
Y
Itakura
Takao
T
D000818:Animals; D001483:Base Sequence; D002347:Carps / Q000235:genetics*; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification*; D029941:Fish Proteins / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic; D005880:Gills / Q000201:enzymology*; D008748:Methylcholanthrene; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D029941:Fish Proteins; D008748:Methylcholanthrene; D003577:Cytochrome P-450 Enzyme System