Molecular Cloning and Functional Analysis of Cynomolgus Monkey CYP1A2

Abstract

Complementary DNA fragments encoding cynomolgus monkey CYP1A2 were amplified by the reverse transcriptase–polymerase chain reaction (RT-PCR) method from the liver total RNA of a 3-methylcholanthrene (3-MC)-treated cynomolgus monkey. The nucleotide sequence determined was 1630 bp long and contained an open reading frame for a polypeptide of 516 residues. The nucleotide and the deduced amino acid sequences of cynomolgus monkey CYP1A2 showed 95.1 and 92.8% identities to those of human CYP1A2, respectively. The level of CYP1A2 mRNA in the liver of untreated cynomolgus monkey was very low. Treatment with 3-MC increased it. Still, it was one-fortieth that of CYP1A1. Cynomolgus monkey CYP1A2 expressed in recombinant yeasts activated 2-amino-3-methylimidazo[4,5- f]quinoline (IQ) and 2-amino-3,8-dimethylimidazo[4,5- f]quinoxaline (MeIQx) at efficient rates in the umu mutagenicity test. This cytochrome P450 (CYP) also activated 2-amino-1-methyl-6-phenylimidazo[4,5- b]pyridine (PhIP), but less efficiently. These results indicate that cynomolgus monkeys have a functionally active CYP1A2 gene, but its expression level is very low in the liver of untreated cynomolgus monkeys.