An alternatively spliced cytochrome P4501A1 in human brain fails to bioactivate polycyclic aromatic hydrocarbons to DNA-reactive metabolites.
CYP1A1, a cytochrome P450 enzyme, metabolizes polycyclic aromatic hydrocarbons to genotoxic metabolite(s) that bind to DNA and initiate carcinogenesis. RT-PCR amplification of the complete open reading frame of CYP1A1 generated an amplicon of 1593 bp having deletion of 87 bp of exon-6 that translated into functional P450 enzyme. Unlike wild type CYP1A1, exon 6 del CYP1A1 did not metabolize polycyclic aromatic hydrocarbons such as, benzo(a)pyrene to genotoxic, ultimate carcinogens that form DNA adducts. Exon 6 del CYP1A1 metabolized ethoxyresorufin (the classical substrate for CYP1A1) less efficiently compared with wild type CYP1A1 while pentoxy and benzyloxyresorufin (classical substrates for CYP2B) were dealkylated more efficiently. In silico docking showed alteration of the substrate access channel in exon 6 del CYP1A1 such that benzo(a)pyrene does not bind in any orientation that would permit the formation of carcinogenic metabolites. Genotyping revealed that the splice variant was not generated due to differences in genomic DNA sequence and the variant was present only in brain but not in liver, kidney, lung, or heart from the same individual. We provide evidence that unique P450 enzymes, generated by alternate splicing in a histiospecific manner can modify genotoxic potential of carcinogens such as benzo(a)pyrene by altering their biotransformation pathway.
Kommaddi
Reddy P
RP
Division of Molecular and Cellular Neuroscience, National Brain Research Centre, Nainwal Mode, Manesar, Haryana, India.
Turman
Cheri M
CM
Moorthy
Bhagavatula
B
Wang
Lihua
L
Strobel
Henry W
HW
Ravindranath
Vijayalakshmi
V
D000293:Adolescent; D000328:Adult; D000368:Aged; D017398:Alternative Splicing / Q000235:genetics*; D001564:Benzo(a)pyrene / Q000378:metabolism; D001711:Biotransformation / Q000235:genetics; D001921:Brain / Q000201:enzymology*; D002273:Carcinogens / Q000378:metabolism; D002648:Child; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics*; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D018736:DNA Adducts / Q000378:metabolism; D004249:DNA Damage / Q000235:genetics; D005260:Female; D006801:Humans; D008658:Inactivation, Metabolic / Q000235:genetics; D008297:Male; D008875:Middle Aged; D009154:Mutation / Q000235:genetics; D010078:Oxazines / Q000378:metabolism; D011084:Polycyclic Aromatic Hydrocarbons / Q000378:metabolism*; D011110:Polymorphism, Genetic / Q000235:genetics*
D002273:Carcinogens; D018736:DNA Adducts; D010078:Oxazines; D011084:Polycyclic Aromatic Hydrocarbons; C095911:polycyclic aromatic hydrocarbons-DNA adduct; D001564:Benzo(a)pyrene; C007649:ethoxyresorufin; D004247:DNA; D019363:Cytochrome P-450 CYP1A1