Detecting ligands and dissecting nuclear receptor-signaling pathways using recombinant strains of the yeast Saccharomyces cerevisiae.
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.
Fox
Jennifer E
JE
Center for Ecology and Evolutionary Biology, University of Oregon, Eugene, Oregon 97403, USA.
Burow
Matthew E
ME
McLachlan
John A
JA
Miller
Charles A
CA
D004271:DNA, Fungal / Q000235:genetics*; D004274:DNA, Recombinant / Q000235:genetics*; D005818:Genetic Engineering; D008024:Ligands; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D015398:Signal Transduction
D004271:DNA, Fungal; D004274:DNA, Recombinant; D008024:Ligands; D018160:Receptors, Cytoplasmic and Nuclear