Metabolic activation of aromatic hydrocarbons in purified rat liver nuclei: induction of enzyme activities and binding to DNA with and without monooxygenase-catalyzed formation of active oxygen.
Purified rat liver nuclei covalently bound low levels of seven aromatic [14C]hydrocarbons to nuclear DNA. Induction with 3-methylcholanthrene increased the binding of six carcinogenic hydorcarbons, but did not raise the level of binding of noncarcinogenic anthracence. Removal of the nuclear envelope by Triton N-101 eliminated binding and aryl hydrocarbon hydroxylase activities and cytochrome P-450 from the nuclei. Binding of two of two strong carcinogens, benzo[a]pyrene and 7,12-dimethylbenz[a]anthracene, to nuclear DNA was compared to the levels of aryl hydrocarbon hydroxylase and cytochrome P-450 in nuclei from uninduced and benz[a]anthracene-, 3-methylcholanthrene-, and phenobarbital-induced rats. Microsomal hydroxylase and cytochrome P-450 were also assayed. Induction with 3-methylcholanthrene gave the largest increases in nuclear activities: 11 times as much hydroxylase, 6 times as much cytochrome P-450, and 4 times as much binding of both hydrocarbons. Benz[a]anthracene and phenobarbital induced these nuclear activities 0- to 4-fold. In the presence of added NADPH, binding of benzol[a]pyrene to DNA by nuclei increased rapidly for at least 20 min. When NADPH was not added, the reaction stopped at a low level in 5 min. When CO was bubbled through the reaction mixture with or without added NADPH, binding of benzo[a]pyrene and 7,12-dimethylbenz[a]anthracene was partially inhibited, indicating that cytochrome P-450 plays a role in this activation. Since no nuclear hydroxylase activity was seen without added NADPH or in the presence of CO, activation and subsequent binding of hydrocarbons to nuclear DNA, at least in part, does not require the activated oxygen used in monooxygenase reactions.
D015127:9,10-Dimethyl-1,2-benzanthracene / Q000378:metabolism; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism; D001551:Benz(a)Anthracenes / Q000494:pharmacology; D001580:Benzopyrenes / Q000378:metabolism; D001665:Binding Sites; D002467:Cell Nucleus / Q000201:enzymology / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D004790:Enzyme Induction; D008099:Liver / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology; D008862:Microsomes, Liver / Q000201:enzymology; D010634:Phenobarbital / Q000494:pharmacology; D011083:Polycyclic Compounds / Q000378:metabolism*; D051381:Rats
D001551:Benz(a)Anthracenes; D001580:Benzopyrenes; D011083:Polycyclic Compounds; D008748:Methylcholanthrene; D015127:9,10-Dimethyl-1,2-benzanthracene; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; D010634:Phenobarbital