Complementary DNA cloning, sequence analysis and differential organ expression of beta-naphthoflavone-inducible cytochrome P4501A in Atlantic salmon (Salmo salar).
Cytochrome p4501A induction and subsequent enzyme expression is used as a biomarker for exposure to aryl hydrocarbon receptor active contaminants in fish and other species. In the present study, CYP1A cDNA (1912 bp, GenBank accession number AF364076) was cloned, sequenced and characterized from the liver of a beta-naphthoflavone (betaNF)-treated teleost, Atlantic salmon (Salmo salar), by reverse-transcriptase polymerase chain reaction (RT-PCR). The salmon CYP1A sequence contained a 5'-flanking region of 99 bp, an open reading frame of 1566 bp that encodes a 521 amino acid protein, a stop codon, and a 3'-untranslated region of 346 bp, and a single polyadenylated signal. The theoretical molecular mass and isoelectric point was 58.6 kDa and 6.17, respectively. Comparison of the deduced amino acid sequence of salmon cDNA with reported CYP1A genes showed identities of 73-88% with fish CYP1A, 51-54% with mammalian CYP1A1, 47-51% with mammalian CYP1A2 and 54% with frog p450. Phylogenetic analysis showed that salmon CYP1A clustered in the tree with rainbow trout CYP1A1 and eel CYP1A sequences. CYP1A mRNA induction in beta-naphthoflavone-treated salmon showed differential organ expression with a distinct single transcript pattern. A new specific molecular tool has been developed for the monitoring of environmental pollution using CYP1A mRNA from salmon as a biomarker.
Arukwe
Augustine
A
Great Lakes Institute for Environmental Research, University of Windsor, 401 Sunset Avenue, Windsor, Ontario, Canada N9B 3P4. arukwe@uwindsor.ca
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D019363:Cytochrome P-450 CYP1A1 / Q000096:biosynthesis* / Q000235:genetics; D018076:DNA, Complementary / Q000096:biosynthesis* / Q000235:genetics; D004790:Enzyme Induction / Q000187:drug effects / Q000502:physiology; D005260:Female; D008297:Male; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics; D010802:Phylogeny; D020317:Salmo salar / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA / Q000379:methods*; D017386:Sequence Homology, Amino Acid; D019324:beta-Naphthoflavone / Q000494:pharmacology*
D018076:DNA, Complementary; D019324:beta-Naphthoflavone; D019363:Cytochrome P-450 CYP1A1