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Related Experiment Video

Updated: Jun 16, 2026

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

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

Published on: November 15, 2013

An orphan nuclear receptor finds a home.

Alan R Kimmel¹

¹Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-8028, USA. ark1@helix.nih.gov

|February 4, 2010

Summary

Retinoic acid receptor-related orphan receptor alpha (RORalpha) suppresses Wnt/beta-catenin signaling. This Wnt5a- and PKC-dependent suppression occurs without affecting beta-catenin stabilization or promoter recruitment.

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

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Production of Nurr-1 Specific Polyclonal Antibodies Free of Cross-reactivity Against Its Close Homologs, Nor1 and Nur77

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Published on: August 17, 2015

Area of Science:

Molecular biology
Cell signaling
Gene regulation

Background:

The Wnt/beta-catenin pathway is crucial for cellular processes, including development and cancer.
RORalpha is a nuclear receptor implicated in various physiological functions.
Cross-talk between nuclear receptors and Wnt signaling is increasingly recognized.

Purpose of the Study:

To investigate the role of RORalpha in regulating Wnt/beta-catenin signaling.
To elucidate the mechanism by which RORalpha influences this pathway.

Main Methods:

Cell-based assays to measure Wnt/beta-catenin activity.
Western blotting to assess protein levels and stabilization.
Reporter assays to evaluate promoter recruitment.

Main Results:

RORalpha was found to suppress Wnt/beta-catenin signaling.
This suppression was dependent on Wnt5a and protein kinase C (PKC).
RORalpha's action did not involve altering beta-catenin stabilization or its recruitment to target gene promoters.

Conclusions:

RORalpha acts as a negative regulator of Wnt/beta-catenin signaling.
The mechanism involves Wnt5a and PKC, independently of canonical beta-catenin stabilization.
This finding reveals a novel regulatory interaction impacting Wnt pathway activity.