Quantitative Interactor Screening with next-generation Sequencing (QIS-Seq) identifies Arabidopsis thaliana MLO2 as a target of the Pseudomonas syringae type III effector HopZ2

BMC Genomics

Research Article

Pauline Fung¹  Yunchen Gong¹  Hardeep Nahal¹  Janet Wan²  Rachel Ford²  Pauline W Wang³  Darrell Desveaux³  David S Guttman³  Jennifer D Lewis⁴ 

[1] Centre for the Analysis of Genome Evolution and Function, University of Toronto, M5S 3B2, Toronto, ON, Canada;Department of Cell and Systems Biology, University of Toronto, M5S 3B2, Toronto, ON, Canada;Department of Cell and Systems Biology, University of Toronto, M5S 3B2, Toronto, ON, Canada;Centre for the Analysis of Genome Evolution and Function, University of Toronto, M5S 3B2, Toronto, ON, Canada;Department of Cell and Systems Biology, University of Toronto, M5S 3B2, Toronto, ON, Canada;Plant Gene Expression Center, USDA, 800 Buchanan St., 94710, Albany, CA, USA;

关键词: Next-generation sequencing;    yeast two-hybrid;    high-throughput screening;    Arabidopsis;    Pseudomonas syringae;    type III effector;    MLO2;    HopZ;   

DOI  :  10.1186/1471-2164-13-8

 received in 2011-09-16, accepted in 2012-01-09,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundIdentification of protein-protein interactions is a fundamental aspect of understanding protein function. A commonly used method for identifying protein interactions is the yeast two-hybrid system.ResultsHere we describe the application of next-generation sequencing to yeast two-hybrid interaction screens and develop Quantitative Interactor Screen Sequencing (QIS-Seq). QIS-Seq provides a quantitative measurement of enrichment for each interactor relative to its frequency in the library as well as its general stickiness (non-specific binding). The QIS-Seq approach is scalable and can be used with any yeast two-hybrid screen and with any next-generation sequencing platform. The quantitative nature of QIS-Seq data make it amenable to statistical evaluation, and importantly, facilitates the standardization of experimental design, data collection, and data analysis. We applied QIS-Seq to identify the Arabidopsis thaliana MLO2 protein as a target of the Pseudomonas syringae type III secreted effector protein HopZ2. We validate the interaction between HopZ2 and MLO2 in planta and show that the interaction is required for HopZ2-associated virulence.ConclusionsWe demonstrate that QIS-Seq is a high-throughput quantitative interactor screen and validate MLO2 as an interactor and novel virulence target of the P. syringae type III secreted effector HopZ2.

【 授权许可】

Unknown   
© Lewis et al; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.