15 Two-Hybrid Analysis of Protein-Protein Interactions in Yeast

Abstract

Publisher Summary This chapter discusses two-hybrid analysis of protein–protein interactions in yeast. Protein–protein interactions underpin a wide range of complex cellular processes and in many cases make a critical contribution to the regulation of these processes. A key question in the study of the cellular function of any novel protein is to identify other proteins with which it interacts. There are many physical, biochemical, and immunological methods available to identify and quantify such interactions. These have been complemented by genetic-based methods of which one—the yeast two-hybrid system—was developed as a sensitive genetic tool to identify protein–protein interactions in vivo. The impact of the two-hybrid method for defining and characterizing protein–protein interactions has been remarkable; although it represents only one of the many available methods for studying such interactions, its application is enhanced considerably by an explosion in genome sequence information. The chapter describes the background to the two-hybrid system and the properties of the basic elements used to define interactions using the two-hybrid system. It also reviews some of the potential future developments of this technology.