Regulation of mouse cytochrome P3-450 by the Ah receptor. Studies with a P3-450 cDNA clone.
The Ah locus in the C57BL/6N mouse regulates at least two cytochrome P-450 gene products, termed in the mouse P1-450 and P3-450; these two enzymes are so named because each is responsible for the highest turnover number for the substrates benzo[a]pyrene and acetanilide, respectively. A cDNA library was prepared in pBR322 from sucrose gradient fractionated total liver poly(A+)-enriched RNA (approximately 20 S) from 2,3,7,8-tetrachlorodibenzo-p-dioxin- (TCDD) treated C57BL/6N (Ahb/Ahb) mice. Differential colony hybridization screening, with [32P]cDNA probes derived from total liver mRNA of both TCDD-treated and control C57BL/6N mice, yielded pP(3)450-21 (1710 base pair) and pP(1)450-57 (1770 base pair) cDNA clones. pP(1)450-57 was found to have 690 base pairs 5'-ward of the original P1-450 cDNA cloned in this laboratory. Restriction maps of pP(3)450-21 and pP(1)450-57 are markedly different and clearly are derived from separate genes. By means of hybridization-translation-arrest experiments, anti-(P3-450) precipitates the translation product (Mr approximately equal to 55000) of mRNA specifically hybridizing to pP(3)450-21. It is also shown that hybridization-translation-arrest experiments using polyclonal antibodies are not specific for proof of a P-450 cDNA clone. pP(3)450-21 was used to probe liver mRNA from Ahb/Ahb, Ahb/Ahd, and Ahd/Ahd mice treated with 3-methylcholanthrene, beta-naphthoflavone, aroclor 1254, isosafrole, low TCDD, or high TCDD. These genetic data rigorously demonstrate control of the P3-450 (20S) mRNA induction process by the Ah receptor. pP(3)450-21 fragments hybridized to TCDD-induced C57BL/6N mRNA and to a portion of the cloned 5' end of the P1-450 gene from a mouse MOPC 41 plasmacytoma library.(ABSTRACT TRUNCATED AT 250 WORDS)
D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D004247:DNA; D004274:DNA, Recombinant; D005260:Female; D007163:Immunosorbent Techniques; D008099:Liver / Q000032:analysis; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009693:Nucleic Acid Hybridization; D010954:Plasmacytoma / Q000235:genetics; D011083:Polycyclic Compounds / Q000494:pharmacology; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism
D004274:DNA, Recombinant; D011083:Polycyclic Compounds; D012333:RNA, Messenger; D004247:DNA; D003577:Cytochrome P-450 Enzyme System