A third genetic locus affecting the Ah (dioxin) receptor.
We have isolated a new benzo(a)pyrene-resistant clone, c35, of the mouse hepatoma line, Hepa-1. Cytochrome P1-450 mRNA and P1-450-dependent aryl hydrocarbon hydroxylase (AHH) activity are no longer inducible by 2,3,7,8-tetrachlorodibenzo-p-dioxin in c35. The phenotype of c35 is unstable in long-term culture. A subclone, c35-1, having partially restored AHH inducibility, was studied in detail. The concentration of dioxin required to give half-maximal induction of AHH activity was 16-fold greater in c35-1 than in Hepa-1. Scatchard analysis showed that c35-1 contains reduced levels of the Ah (dioxin) receptor, which mediates induction of P1-450, but that the affinity of the receptor for dioxin is unaltered. In vivo assays confirmed that c35-1 possesses reduced levels of receptor but showed that it is even more severely affected in nuclear translocation of the receptor. Somatic cell hybridization experiments demonstrated that c35 is recessive and belongs to a new, third complementation group of mutants defective in Ah receptor activity. We propose that c35 is mutated either in the ligand-binding Ah receptor polypeptide or in another polypeptide required for receptor function and that in c35-1 partial reversion has occurred to generate a polypeptide which is still impaired in its role in promoting nuclear translocation and/or DNA binding.
Karenlampi
S O
SO
Laboratory of Biomedical and Environmental Sciences, School of Public Health, University of California, Los Angeles 90024.
Legraverend
C
C
Gudas
J M
JM
Carramanzana
N
N
Hankinson
O
O
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002999:Clone Cells; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D004147:Dioxins / Q000378:metabolism*; D004790:Enzyme Induction; D005796:Genes; D007700:Kinetics; D008114:Liver Neoplasms, Experimental / Q000378:metabolism; D051379:Mice; D009154:Mutation; D009251:NADPH-Ferrihemoprotein Reductase / Q000235:genetics / Q000378:metabolism; D000072317:Polychlorinated Dibenzodioxins / Q000378:metabolism* / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D018336:Receptors, Aryl Hydrocarbon; D011955:Receptors, Drug / Q000235:genetics*
D004147:Dioxins; D000072317:Polychlorinated Dibenzodioxins; D012333:RNA, Messenger; D018336:Receptors, Aryl Hydrocarbon; D011955:Receptors, Drug; D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; D009251:NADPH-Ferrihemoprotein Reductase