Calcyclin (S100A6) expression is stimulated by agents evoking oxidative stress via the antioxidant response element.
Calcyclin (S100A6) is a cell-specific, calcium binding protein of the S100 family whose expression is augmented in many types of cancer. By means of luciferase activity assays, RT-PCR and Northern blot hybridization, we established that transcription of S100A6 gene is increased by agents known to evoke oxidative stress. Mutation of the antioxidant response element (ARE) located at position -290/-281 of the calcyclin gene promoter, and overlapping the E-box sequence recognized by the upstream stimulatory factor (USF), led to inhibition of calcyclin gene promoter activity stimulated by cadmium ions. Electrophoretic mobility shift assays (EMSA) with the -302/-260 calcyclin gene promoter fragment revealed, apart from USF binding, the presence of another protein complex (N) shown by competitive EMSA to be bound to ARE. DNA affinity chromatography followed by Western blot showed the binding of Nrf2 transcription factor to the immobilized calcyclin gene promoter fragment and concomitant appearance of complex N in EMSA of the eluted fractions. The results indicate that agents evoking oxidative stress activate calcyclin gene via the ARE sequence in its promoter.
Leśniak
Wiesława
W
Department of Molecular and Cellular Neurobiology, Nencki Institute of Experimental Biology, 3 Pasteur Street, 02-093 Warsaw, Poland. w.lesniak@nencki.gov.pl
Szczepańska
Anna
A
Kuźnicki
Jacek
J
D000975:Antioxidants / Q000494:pharmacology; D001483:Base Sequence; D019256:Cadmium Chloride / Q000494:pharmacology; D018797:Cell Cycle Proteins / Q000235:genetics*; D002478:Cells, Cultured; D003474:Curcumin / Q000494:pharmacology; D016213:Cyclins / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism; D024721:E-Box Elements / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation; D016877:Oxidants / Q000494:pharmacology*; D018384:Oxidative Stress / Q000235:genetics*; D011401:Promoter Regions, Genetic; D020218:Response Elements / Q000187:drug effects* / Q000235:genetics; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism; D051720:Upstream Stimulatory Factors
D000975:Antioxidants; D018797:Cell Cycle Proteins; D016213:Cyclins; D004268:DNA-Binding Proteins; D016877:Oxidants; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins; D014157:Transcription Factors; C497181:USF1 protein, human; D051720:Upstream Stimulatory Factors; C055351:S100A6 protein, human; D006861:Hydrogen Peroxide; D003474:Curcumin; D019256:Cadmium Chloride