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Reverse Yeast Two-hybrid System to Identify Mammalian Nuclear Receptor Residues that Interact with Ligands and/or Antagonists
10:51

Reverse Yeast Two-hybrid System to Identify Mammalian Nuclear Receptor Residues that Interact with Ligands and/or Antagonists

Published on: November 15, 2013

Nuclear receptor-coregulator interaction profiling identifies TRIP3 as a novel peroxisome proliferator-activated

Arjen Koppen1, Rene Houtman, Dirk Pijnenburg

  • 1Department of Metabolic and Endocrine Diseases, University Medical Centre Utrecht, Utrecht, The Netherlands.

Molecular & Cellular Proteomics : MCP
|July 15, 2009
PubMed

Insights

Nuclear receptors (NRs) regulate gene transcription with coregulators. This study profiles NR-coregulator interactions, identifying TRIP3 as a novel regulator of PPARgamma, aiding drug discovery.

Area of Science:

  • Molecular Biology
  • Endocrinology
  • Drug Discovery

Background:

  • Nuclear receptors (NRs) are crucial drug targets involved in homeostasis and diseases like cancer and diabetes.
  • NRs function as ligand-dependent transcription factors, interacting with coregulators (coactivators/corepressors) to control gene expression.
  • Specific alpha-helical motifs (LXXLL/LXXXL) mediate NR-coregulator binding, but interaction specificity remains poorly understood.

Purpose of the Study:

  • To comprehensively map nuclear receptor-coregulator interactions.
  • To characterize ligand-specific binding profiles of peroxisome proliferator-activated receptor gamma (PPARgamma).
  • To identify novel regulators of NR-mediated transcriptional activity.

Main Methods:

  • Utilized a microarray approach to analyze interactions between NRs and coregulator-derived peptides.
  • Generated ligand-specific interaction profiles for PPARgamma using an agonist (rosiglitazone), antagonist (GW9662), and selective modulator (telmisartan).
  • Characterized interaction profiles of PPARgamma mutants and isotypes (PPARalpha, -beta/delta).

Main Results:

  • Developed ligand-specific interaction profiles for PPARgamma, revealing differential coregulator binding.
  • Identified TRIP3 as a novel coregulator of PPARgamma.
  • Demonstrated TRIP3's role in regulating PPARgamma-mediated adipocyte differentiation.

Conclusions:

  • NR-coregulator interaction profiling provides a valuable tool for understanding NR function.
  • This approach can facilitate the discovery of novel NR regulators and therapeutic targets.
  • Findings highlight the potential of interaction profiling in drug development and biological discovery.

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