Development and application of a DNA microarray-based yeast two-hybrid system
Authors
- B. Suter
- J.F. Fontaine
- R. Yildirimman
- T. Raskó
- M.H. Schaefer
- A. Rasche
- P. Porras
- B.M. Vázquez-Álvarez
- J. Russ
- K. Rau
- R. Foulle
- M. Zenkner
- K. Saar
- R. Herwig
- M.A. Andrade-Navarro
- E.E. Wanker
Journal
- Nucleic Acids Research
Citation
- Nucleic Acids Res 41 (3): 1496-507
Abstract
The yeast two-hybrid (Y2H) system is the most widely applied methodology for systematic protein-protein interaction (PPI) screening and the generation of comprehensive interaction networks. We developed a novel Y2H interaction screening procedure using DNA microarrays for high-throughput quantitative PPI detection. Applying a global pooling and selection scheme to a large collection of human open reading frames, proof-of-principle Y2H interaction screens were performed for the human neurodegenerative disease proteins huntingtin and ataxin-1. Using systematic controls for unspecific Y2H results and quantitative benchmarking, we identified and scored a large number of known and novel partner proteins for both huntingtin and ataxin-1. Moreover, we show that this parallelized screening procedure and the global inspection of Y2H interaction data are uniquely suited to define specific PPI patterns and their alteration by disease-causing mutations in huntingtin and ataxin-1. This approach takes advantage of the specificity and flexibility of DNA microarrays and of the existence of solid-related statistical methods for the analysis of DNA microarray data, and allows a quantitative approach toward interaction screens in human and in model organisms.