FCCC LOGO Faculty Publications
Li Y , Jaiswal AK
Regulation of Human Nad(P)H-Quinone Oxidoreductase Gene - Role of Ap1 Binding-Site Contained within Human Antioxidant Response Element
Journal of Biological Chemistry. 1992 Jul 25;267(21) :15097-15104
PMID: ISI:A1992JF08800087   
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Deletion mutagenesis and transfection studies into hepatic (mouse hepatoma (Hepa- 1) and human hepatoblastoma (Hep-G2)) and nonhepatic (HeLa) cells indicated that high levels of expression of the human NAD(P)H:quinone oxidoreductase gene in tumor cells and its induction by beta-naphthoflavone and 3-(2)- tert-butyl-4-hydroxyanisole are mediated by human antioxidant response element (hARE) located in the region between -470 and -445. The hARE, when attached to the thymidine kinase promoter and transfected into several mammalian cells, expressed high levels of the chloramphenicol acetyltransferase gene that was inducible by beta-naphthonavone and 3-(2)-tert-butyl-4- hydroxyanisole. Nucleotide sequence analysis of the hARE revealed the presence of a recognition site for binding to the AP1 protein. Mutation of the AP1 binding site located within the hARE resulted in the loss of expression and induction upon transfection into various cell types. Band shift and competition assays with hARE and nuclear extracts from control and beta-naphthoflavone-treated Hepa-1, Hep-G2 and HeLa cells indicated specific interaction of regulatory protein(s) to the hARE. The supershift assays using antibodies against specific proteins of the AP1 family identified Jun-D and c-Fos as two members in the hARE-protein complex observed in band shift assays.
English Article JF088 J BIOL CHEM