The yeast two-hybrid and related methods as powerful tools to study plant cell signalling

One basic property of proteins is their ability to specifically target and form non-covalent complexes with other proteins. Such protein-protein interactions play key roles in all biological processes, extending from the formation of cellular macromolecular structures and enzymatic complexes to the regulation of signal transduction pathways. Identifying and characterizing protein interactions and entire interaction networks (interactomes) is therefore prerequisite to understand these processes on a molecular and biophysical level. Since its original description in 1989, the yeast two-hybrid system has been extensively used to identify protein-protein interactions from many different organisms, thus providing a convenient mean to both screen for proteins that interact with a protein of interest and to characterize the known interaction between two proteins. In these years the technique has improved to overcome the limitations of the original assay, and many efforts have been made to scale up the technique and to adapt it to large scale studies. In addition, variations have been introduced to enlarge the range of proteins and interactors that can be assayed by hybrid-based approaches. Several groups studying molecular mechanisms that underlie plant cell signal transduction pathways have successfully used the yeast two-hybrid system or related methods. In this review we provide a brief description of the technology, attempt to point out some of the pitfalls and benefits of the different systems that can be employed, and mention some of the areas, within the plant cell signalling field, where hybrid-based interaction assays have been particularly informative.