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Related Experiment Videos

Zinc rapidly induces a metal response element-binding factor.

M Czupryn1, W E Brown, B L Vallee

  • 1Center for Biochemical and Biophysical Sciences and Medicine, Harvard Medical School, Boston, MA 02115.

Proceedings of the National Academy of Sciences of the United States of America
|November 1, 1992
PubMed
Summary

This study identifies two distinct DNA-binding factors (BF1 and BF2) that interact with metal regulatory elements (MREs) controlling metallothionein gene transcription. Metal exposure induces a second factor (BF2), essential for MRE binding.

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Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Toxicology

Background:

  • Metallothionein (MT) gene transcription is activated by metals via specific promoter sequences called metal regulatory elements (MREs).
  • Understanding the proteins that bind MREs is crucial for elucidating metal-induced gene expression.
  • Human cell lines provide a model system to study these interactions.

Purpose of the Study:

  • To identify and characterize the protein factors that bind to the human MREa (hMREa) sequence.
  • To investigate how metal exposure influences the binding of these factors to hMREa.
  • To elucidate the role of metals in the interaction between these factors and DNA.

Main Methods:

  • Electrophoretic mobility-shift assays (EMSA) were used to detect protein-DNA complexes.

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  • DNA-binding protein blotting and UV crosslinking identified and characterized the binding factors.
  • Electroelution experiments confirmed the metal-dependent nature of the interaction.

    • Main Results:

    • A single hMREa-binding complex (BF1, ~86 kDa) was detected in control cells.
    • Metal treatment (zinc, cadmium, copper) induced a second complex (BF2, ~2x28 kDa) in all cell lines studied.
    • EDTA and o-phenanthroline inhibited binding, confirming the essential role of metals.

    Conclusions:

    • Two distinct MRE-binding factors, BF1 and BF2, are involved in metallothionein gene regulation.
    • Metal-induced BF2 formation or binding is critical for MRE interaction.
    • Metal ions are essential components for the DNA-binding activity of these factors.