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

Nucleic Acids Res.2013 Feb 1;41(3):1496-507. Epub 2012 Dec 28
Bernhard Suter 1 , Jean-Fred Fontaine , Reha Yildirimman , Tamás Raskó , Martin H Schaefer , Axel Rasche , Pablo Porras , Blanca M Vázquez-Álvarez , Jenny Russ , Kirstin Rau , Raphaele Foulle , Martina Zenkner , Kathrin Saar , Ralf Herwig , Miguel A Andrade-Navarro , Erich E Wanker
Bernhard Suter 1 , Jean-Fred Fontaine , Reha Yildirimman , Tamás Raskó , Martin H Schaefer , Axel Rasche , Pablo Porras , Blanca M Vázquez-Álvarez , Jenny Russ , Kirstin Rau , Raphaele Foulle , Martina Zenkner , Kathrin Saar , Ralf Herwig , Miguel A Andrade-Navarro , Erich E Wanker
+ et al

[No authors listed]

Author information
  • 1 Max Delbrueck Center for Molecular Medicine, Berlin 13125, Germany. bernhard.suter@quintarabio.com

摘要


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.