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Related Experiment Videos

Modulation of hormone-dependent glucocorticoid receptor function using a tetracycline-regulated expression system

P Wei1, Y I Ahn, P R Housley

  • 1Department of Biochemistry and Molecular Biology, Louisiana State University Medical Center, New Orleans 70112, USA.

The Journal of Steroid Biochemistry and Molecular Biology
|May 6, 1998
PubMed
Summary

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Researchers developed a novel cell line for conditional glucocorticoid receptor (GR) expression. This system allows precise control over GR levels, enabling detailed study of its gene regulation functions.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Endocrinology

Background:

  • The glucocorticoid receptor (GR) is a crucial transcription factor mediating glucocorticoid effects.
  • Understanding GR's dual role in gene regulation (activation and repression) is vital for biological and medical research.
  • Existing models may not offer sufficient control over intracellular GR concentrations.

Purpose of the Study:

  • To create a novel mammalian cell line with tetracycline-regulated expression of rat GR (rGR).
  • To establish a system for precisely manipulating intracellular rGR levels.
  • To investigate the dose-dependent effects of GR on gene transactivation and transrepression.

Main Methods:

  • Development of the E8.2/GR3 cell line from GR null mouse fibroblasts using a tetracycline-regulated expression system.

Related Experiment Videos

  • Manipulation of intracellular rGR concentration by varying tetracycline (Tc) levels.
  • Assay of dexamethasone (DEX)-dependent transactivation and transrepression activities modulated by rGR levels.

    • Main Results:

    • Conditional rGR expression was successfully established and modulated by Tc concentration.
    • DEX-dependent transactivation and transrepression were strictly dependent on rGR levels.
    • DEX induced rGR-dependent morphological changes in the cells.
    • Half-maximal transactivation and transrepression required similar Tc concentrations but different DEX concentrations.

    Conclusions:

    • The E8.2/GR3 cell line provides a powerful, controllable system for studying GR functions.
    • This model allows for detailed examination of GR's role in gene expression across a range of intracellular receptor concentrations.
    • The findings highlight the complex, concentration-dependent nature of GR-mediated gene regulation.