Yeast two-hybrid, a powerful tool for systems biology.
A key property of complex biological systems is the presence of interaction networks formed by its different components, primarily proteins. These are crucial for all levels of cellular function, including architecture, metabolism and signalling, as well as the availability of cellular energy. Very stable, but also rather transient and dynamic protein-protein interactions generate new system properties at the level of multiprotein complexes, cellular compartments or the entire cell. Thus, interactomics is expected to largely contribute to emerging fields like systems biology or systems bioenergetics. The more recent technological development of high-throughput methods for interactomics research will dramatically increase our knowledge of protein interaction networks. The two most frequently used methods are yeast two-hybrid (Y2H) screening, a well established genetic in vivo approach, and affinity purification of complexes followed by mass spectrometry analysis, an emerging biochemical in vitro technique. So far, a majority of published interactions have been detected using an Y2H screen. However, with the massive application of this method, also some limitations have become apparent. This review provides an overview on available yeast two-hybrid methods, in particular focusing on more recent approaches. These allow detection of protein interactions in their native environment, as e.g. in the cytosol or bound to a membrane, by using cytosolic signalling cascades or split protein constructs. Strengths and weaknesses of these genetic methods are discussed and some guidelines for verification of detected protein-protein interactions are provided.
Brückner
Anna
A
INSERM U884, Université Joseph Fourier, Laboratoire de Bioénergétique Fondamentale et Appliquée, 2280 Rue de la Piscine, BP 53, Grenoble Cedex 9, France.
Polge
Cécile
C
INSERM U884, Université Joseph Fourier, Laboratoire de Bioénergétique Fondamentale et Appliquée, 2280 Rue de la Piscine, BP 53, Grenoble Cedex 9, France.
Lentze
Nicolas
N
Dualsystems Biotech AG / Grabenstrasse 11a, 8952 Schlieren, Switzerland.
Auerbach
Daniel
D
Dualsystems Biotech AG / Grabenstrasse 11a, 8952 Schlieren, Switzerland.
Schlattner
Uwe
U
INSERM U884, Université Joseph Fourier, Laboratoire de Bioénergétique Fondamentale et Appliquée, 2280 Rue de la Piscine, BP 53, Grenoble Cedex 9, France.
D013058:Mass Spectrometry; D060066:Protein Interaction Maps; D011506:Proteins / Q000737:chemistry / Q000378:metabolism; D012320:RNA Polymerase III / Q000378:metabolism; D049490:Systems Biology; D020798:Two-Hybrid System Techniques
D011506:Proteins; D012320:RNA Polymerase III