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

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Item Type:Article
Title:Development and application of a DNA microarray-based yeast two-hybrid system
Creators Name:Suter, B. and Fontaine, J.F. and Yildirimman, R. and Raskó, T. and Schaefer, M.H. and Rasche, A. and Porras, P. and Vázquez-Álvarez, B.M. and Russ, J. and Rau, K. and Foulle, R. and Zenkner, M. and Saar, K. and Herwig, R. and Andrade-Navarro, M.A. and Wanker, E.E.
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.
Keywords:Mutation, Nerve Tissue Proteins, Nuclear Proteins, Oligonucleotide Array Sequence Analysis, Open Reading Frames, Protein Interaction Maps, Two-Hybrid System Techniques, Yeasts
Source:Nucleic Acids Research
Publisher:Oxford University Press
Page Range:1496-507
Date:1 February 2013
Official Publication:https://doi.org/10.1093/nar/gks1329
PubMed:View item in PubMed

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