Progress and variations in two-hybrid and three-hybrid technologies

Abstract

The original yeast two-hybrid system and its variants have proven to be effective tools for identification and analysis of protein–protein, protein–DNA and protein–RNA interactions. The two-hybrid assay is being applied to the entire complement of proteins of the yeast Saccharomyces cerevisiae to characterize the network of protein–protein interactions in the eukaryotic cell. The development of nontranscriptional cytosolic and membrane-associated two-hybrid methods has made it possible to detect and examine a number of protein–protein interactions in their normal cellular locations. Small-molecule hybrid systems have been developed which can be used to study protein–ligand interactions and to activate cellular processes by forcing protein associations.