Identification and quantification of a messenger ribonucleic acid induced by polynuclear aromatic hydrocarbons--using a cloned human cytochrome P-450 gene.
We have isolated four overlapping human genomic clones associated with the polynuclear aromatic hydrocarbon-induced form of cytochrome P-450. The form of P-450 most closely associated with polynuclear aromatic hydrocarbons induction has been defined as P1-450. These four overlapping genomic clones span a total of 31.0 X 10(3) base pairs in length with the coding sequence lying in the center of these clones. Translation in vitro of 3-methylcholanthrene-induced mRNA, selected with the human P1-450 genomic clone, detect a protein with Mr 52000, which is immunoprecipitable by the anti-(mouse P1-450) antibody. The isolated human P1-450 genomic clone hybridizes to 3-methylcholanthrene-induced mRNA from monkey liver, benzanthracene and 3-methylcholanthrene-treated human mammary tumor cells (MCF-7), but not to isosafrole-treated human cells. Upon treatment with polynuclear aromatic hydrocarbons there is a positive correlation between induced arylhydrocarbon hydroxylase (flavoprotein-linked monoxygenase) activity and the amount of mRNA that hybridizes to the isolated human genomic clone for P1-450. The size of mRNA, induced from human cells and monkey liver by polynuclear aromatic hydrocarbons, is around 3.3 X 10(3) base pairs, which is the same as the larger of two mRNA induced by polynuclear aromatic hydrocarbons in the inbred strain of mouse (C57BL/6N). Our data also showed that the isolated DNA clone can detect a mRNA size of 3.3 X 10(3) base pairs from phytohemagglutinin-activated, benzanthracene-treated human lymphocytes. Densitometer scanning indicated the presence of a 3.6-fold variation (highest-lowest) in the levels of lymphocyte P1-450 mRNA contents among six individuals studied.
D000818:Animals; D001551:Benz(a)Anthracenes / Q000494:pharmacology; D001579:Benzopyrene Hydroxylase / Q000096:biosynthesis / Q000235:genetics; D002273:Carcinogens / Q000494:pharmacology*; D002460:Cell Line; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000235:genetics*; D004247:DNA; D003979:Diazonium Compounds; D004587:Electrophoresis, Agar Gel; D005796:Genes; D006801:Humans; D006867:Hydrolases / Q000096:biosynthesis / Q000235:genetics; D008214:Lymphocytes / Q000378:metabolism; D008252:Macaca fascicularis; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D011083:Polycyclic Compounds / Q000494:pharmacology*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis*; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D012451:Safrole / Q000494:pharmacology
D001551:Benz(a)Anthracenes; D002273:Carcinogens; D003979:Diazonium Compounds; D011083:Polycyclic Compounds; D012333:RNA, Messenger; C015794:diazobenzyloxymethyl groups; D008748:Methylcholanthrene; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; C030935:benz(a)anthracene; D001579:Benzopyrene Hydroxylase; D006867:Hydrolases; D012451:Safrole; C015959:isosafrole