Detection of a functional xenobiotic response element in a widely employed FoxO-responsive reporter construct.

Anna Eckers, Elisabeth Sauerbier, Anwar Anwar-Mohamed, Ingrit Hamann, Charlotte Esser, Peter Schroeder, Ayman O S El-Kadi, Lars-Oliver Klotz
Author Information
  1. Anna Eckers: Leibniz-Institut für umweltmedizinische Forschung, Düsseldorf, Germany.

Abstract

FHRE-Luc is a promoter reporter construct that is widely used to assess the activity of FoxO (forkhead box, class O) transcription factors. We here demonstrate that this promoter construct responds to exposure of HepG2 human hepatoma cells to known agonists of the aryl hydrocarbon receptor (AhR), 3-methylcholanthrene, benzo(a)pyrene, and 6-formylindolo[3,2-b]carbazole. However, FHRE-Luc activation did not coincide with FoxO DNA binding or changes in Akt-induced FoxO phosphorylation after treatment with AhR agonists. Testing FHRE-Luc deletion constructs and using AhR-deficient cells, we found that FHRE-Luc activation by AhR agonists is due to a functional xenobiotic-response element (XRE) spanning the backbone/insert border of the reporter plasmid. In conclusion, care must be taken when using FHRE-Luc to assess FoxO activity in response to stimuli that potentially interfere with xenobiotic signaling.

MeSH Term

Benzo(a)pyrene
Carbazoles
Forkhead Transcription Factors
Genes, Reporter
Hep G2 Cells
Humans
Luciferases
Methylcholanthrene
Phosphorylation
Receptors, Aryl Hydrocarbon
Response Elements
Signal Transduction
Xenobiotics

Chemicals

6-formylindolo(3,2-b)carbazole
Carbazoles
Forkhead Transcription Factors
Receptors, Aryl Hydrocarbon
Xenobiotics
Benzo(a)pyrene
Methylcholanthrene
Luciferases

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