Teaching the Basics of Nuclear Receptor Action: A Simple Laboratory Exercise Using the Yeast Two-Hybrid Method

Abstract

Objectives. Nuclear receptors (NRs) are ligand-activated transcription factors that regulate development and cellular differentiation, endocrine functions, and metabolism and transport of nutrients and drugs. Understanding of NR action is therefore essential for students of pharmacy and biology. Currently, there are no convenient laboratory exercises that clearly demonstrate the components and mechanisms of NR action. Design. Employing the yeast two-hybrid method, we developed a simple hands-on exercise that demonstrated ligand binding, dose responses, and consequences of ligand binding that are critical for NR function. Assessment. The exercise uses inexpensive, non-radioactive reagents and standard laboratory equipment. The results are directly visible, easy to interpret, and reproducible. The whole exercise can be conducted in a single day. In a real life test, a majority of the students who participated reported that their understanding of NR action had increased. Conclusions. This simple exercise can...