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Related Experiment Videos

Functional antioxidant responsive elements

W W Wasserman1, W E Fahl

  • 1McArdle Laboratory for Cancer Research, University of Wisconsin, 1400 University Avenue, Madison, WI 53706, USA.

Proceedings of the National Academy of Sciences of the United States of America
|May 13, 1997
PubMed
Summary

Chemoprotective compounds induce resistance to carcinogens by increasing protective enzymes. This study identifies crucial sequences within antioxidant-responsive elements (AREs) for gene induction, enhancing our understanding of cellular defense mechanisms.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Toxicology

Background:

  • Chemoprotective compounds enhance cellular resistance to carcinogens.
  • This protection involves increased expression of enzymes like glutathione S-transferases (GST).
  • Antioxidant-responsive elements (AREs) regulate the transcriptional induction of these protective genes.

Purpose of the Study:

  • To identify the complete DNA sequences required for a functional antioxidant-responsive element (ARE).
  • To investigate the role of flanking sequences in ARE-mediated gene induction.
  • To classify AREs based on their induction strength and propose a regulatory model.

Main Methods:

  • Systematic mutational analysis of the murine GST Ya ARE.
  • Introduction of identified nucleotides into a GST-Mu promoter sequence.

Related Experiment Videos

  • GenBank database screening for functional ARE consensus sequences.
  • Electrophoretic mobility-shift assay (EMSA) to assess protein binding.

    • Main Results:

    • Additional nucleotides flanking the ARE core sequence are necessary for functional induction.
    • A functional ARE consensus sequence was identified in 16 genes, including the murine ferritin-L gene.
    • The ferritin-L ARE binds to ARE-binding protein 1 (AREB1).
    • A three-level classification system for AREs was developed based on induction strength.

    Conclusions:

    • Functional AREs require specific flanking sequences in addition to the core motif.
    • AREs represent composite regulatory sites involving interactions of multiple transcription factors.
    • This research provides a deeper understanding of the molecular mechanisms underlying chemoprotection and gene regulation.