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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 quantitativ...
Autores principales: | , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3561971/ https://www.ncbi.nlm.nih.gov/pubmed/23275563 http://dx.doi.org/10.1093/nar/gks1329 |
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author | Suter, Bernhard Fontaine, Jean-Fred Yildirimman, Reha Raskó, Tamás Schaefer, Martin H. Rasche, Axel Porras, Pablo Vázquez-Álvarez, Blanca M. Russ, Jenny Rau, Kirstin Foulle, Raphaele Zenkner, Martina Saar, Kathrin Herwig, Ralf Andrade-Navarro, Miguel A. Wanker, Erich E. |
author_facet | Suter, Bernhard Fontaine, Jean-Fred Yildirimman, Reha Raskó, Tamás Schaefer, Martin H. Rasche, Axel Porras, Pablo Vázquez-Álvarez, Blanca M. Russ, Jenny Rau, Kirstin Foulle, Raphaele Zenkner, Martina Saar, Kathrin Herwig, Ralf Andrade-Navarro, Miguel A. Wanker, Erich E. |
author_sort | Suter, Bernhard |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3561971 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-35619712013-02-01 Development and application of a DNA microarray-based yeast two-hybrid system Suter, Bernhard Fontaine, Jean-Fred Yildirimman, Reha Raskó, Tamás Schaefer, Martin H. Rasche, Axel Porras, Pablo Vázquez-Álvarez, Blanca M. Russ, Jenny Rau, Kirstin Foulle, Raphaele Zenkner, Martina Saar, Kathrin Herwig, Ralf Andrade-Navarro, Miguel A. Wanker, Erich E. Nucleic Acids Res Computational Biology 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. Oxford University Press 2013-02 2012-12-26 /pmc/articles/PMC3561971/ /pubmed/23275563 http://dx.doi.org/10.1093/nar/gks1329 Text en © The Author(s) 2012. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Computational Biology Suter, Bernhard Fontaine, Jean-Fred Yildirimman, Reha Raskó, Tamás Schaefer, Martin H. Rasche, Axel Porras, Pablo Vázquez-Álvarez, Blanca M. Russ, Jenny Rau, Kirstin Foulle, Raphaele Zenkner, Martina Saar, Kathrin Herwig, Ralf Andrade-Navarro, Miguel A. Wanker, Erich E. Development and application of a DNA microarray-based yeast two-hybrid system |
title | Development and application of a DNA microarray-based yeast two-hybrid system |
title_full | Development and application of a DNA microarray-based yeast two-hybrid system |
title_fullStr | Development and application of a DNA microarray-based yeast two-hybrid system |
title_full_unstemmed | Development and application of a DNA microarray-based yeast two-hybrid system |
title_short | Development and application of a DNA microarray-based yeast two-hybrid system |
title_sort | development and application of a dna microarray-based yeast two-hybrid system |
topic | Computational Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3561971/ https://www.ncbi.nlm.nih.gov/pubmed/23275563 http://dx.doi.org/10.1093/nar/gks1329 |
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