title: Development and application of a DNA microarray-based yeast two-hybrid system.
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.
authors.affiliation: Max Delbrueck Center for Molecular Medicine, Berlin 13125, Germany. bernhard.suter@quintarabio.com
authors.forename: Bernhard
authors.initials: B
authors.lastname: Suter
authors.forename: Jean-Fred
authors.initials: JF
authors.lastname: Fontaine
authors.forename: Reha
authors.initials: R
authors.lastname: Yildirimman
authors.forename: Tamás
authors.initials: T
authors.lastname: Raskó
authors.forename: Martin H
authors.initials: MH
authors.lastname: Schaefer
authors.forename: Axel
authors.initials: A
authors.lastname: Rasche
authors.forename: Pablo
authors.initials: P
authors.lastname: Porras
authors.forename: Blanca M
authors.initials: BM
authors.lastname: Vázquez-Álvarez
authors.forename: Jenny
authors.initials: J
authors.lastname: Russ
authors.forename: Kirstin
authors.initials: K
authors.lastname: Rau
authors.forename: Raphaele
authors.initials: R
authors.lastname: Foulle
authors.forename: Martina
authors.initials: M
authors.lastname: Zenkner
authors.forename: Kathrin
authors.initials: K
authors.lastname: Saar
authors.forename: Ralf
authors.initials: R
authors.lastname: Herwig
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Andrade-Navarro
authors.forename: Erich E
authors.initials: EE
authors.lastname: Wanker
mesh_terms: D000067696:Ataxin-1; D000067528:Ataxins; D006801:Humans; D000071058:Huntingtin Protein; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016366:Open Reading Frames; D060066:Protein Interaction Maps; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics
chemical_list: C000593749:ATXN1 protein, human; D000067696:Ataxin-1; D000067528:Ataxins; C086055:HTT protein, human; D000071058:Huntingtin Protein; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins