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

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.

Pubmed ID: 23275563

Authors

  • Suter B
  • Fontaine JF
  • Yildirimman R
  • Raskó T
  • Schaefer MH
  • Rasche A
  • Porras P
  • Vázquez-Álvarez BM
  • Russ J
  • Rau K
  • Foulle R
  • Zenkner M
  • Saar K
  • Herwig R
  • Andrade-Navarro MA
  • Wanker EE

Journal

Nucleic acids research

Publication Data

February 1, 2013

Associated Grants

None

Mesh Terms

  • Humans
  • Mutation
  • Nerve Tissue Proteins
  • Nuclear Proteins
  • Oligonucleotide Array Sequence Analysis
  • Open Reading Frames
  • Protein Interaction Maps
  • Two-Hybrid System Techniques
  • Yeasts