Development and application of a DNA microarray-based yeast two-hybrid system

作者：Suter Bernhard^*; Fontaine Jean Fred; Yildirimman Reha; Rasko Tamas; Schaefer Martin H; Rasche Axel; Porras Pablo; Vazquez Alvarez Blanca M; Russ Jenny; Rau Kirstin; Foulle Raphaele; Zenkner Martina; Saar Kathrin; Herwig Ralf; Andrade Navarro Miguel A; Wanker Erich E

来源：Nucleic Acids Research, 2013, 41(3): 1496-1507.

DOI：10.1093/nar/gks1329

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 bench-marking, 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.

• 出版日期2013-2