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Detecting ligands and dissecting nuclear receptor-signaling pathways using recombinant strains of the yeast Saccharomyces cerevisiae

Abstract

This is a general protocol for the identification of natural and xenobiotic ligands of metazoan nuclear receptors (NRs) expressed in yeast. Yeast engineered to express an NR and a response element-driven reporter gene provide a system to detect and quantify ligand-dependent transcriptional activity. Such assays allow researchers to measure different types of ligands and determine dose-dependent activation of NRs. This methodology can also be used to examine the components of signal transduction pathways when conducted with mutant or engineered yeast strains expressing additional proteins or having alternate DNA response elements. This assay typically takes 2–3 d to complete, but most of this time entails cell growth rather than 'hands on' time.

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Figure 1: Typical results seen from experiments using yeast strains expressing the human AhR and Arnt proteins and an RE-driven lacZ reporter plasmid.
Figure 2: Yeast reporter strains identify other proteins involved in NR–ligand-signaling pathways.

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Fox, J., Burow, M., McLachlan, J. et al. Detecting ligands and dissecting nuclear receptor-signaling pathways using recombinant strains of the yeast Saccharomyces cerevisiae. Nat Protoc 3, 637–645 (2008). https://doi.org/10.1038/nprot.2008.33

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