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A Comparative Approach to Characterize the Landscape of Host-Pathogen Protein-Protein Interactions

Abstract : Significant efforts were gathered to generate large-scale comprehensive protein-protein interaction network maps. This is instrumental to understand the pathogen-host relationships and was essentially performed by genetic screenings in yeast two-hybrid systems. The recent improvement of protein-protein interaction detection by a Gaussia luciferase-based fragment complementation assay now offers the opportunity to develop integrative comparative interactomic approaches necessary to rigorously compare interaction profiles of proteins from different pathogen strain variants against a common set of cellular factors. This paper specifically focuses on the utility of combining two orthogonal methods to generate protein-protein interaction datasets: yeast two-hybrid (Y2H) and a new assay, high-throughput Gaussia princeps protein complementation assay (HT-GPCA) performed in mammalian cells. A large-scale identification of cellular partners of a pathogen protein is performed by mating-based yeast two-hybrid screenings of cDNA libraries using multiple pathogen strain variants. A subset of interacting partners selected on a high-confidence statistical scoring is further validated in mammalian cells for pair-wise interactions with the whole set of pathogen variants proteins using HT-GPCA. This combination of two complementary methods improves the robustness of the interaction dataset, and allows the performance of a stringent comparative interaction analysis. Such comparative interactomics constitute a reliable and powerful strategy to decipher any pathogen-host interplays.
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Submitted on : Monday, January 7, 2019 - 10:36:52 AM
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Mandy Muller, Patricia Cassonnet, Michel Favre, Yves Jacob, Caroline Demeret. A Comparative Approach to Characterize the Landscape of Host-Pathogen Protein-Protein Interactions. Journal of visualized experiments : JoVE, JoVE, 2013, pp.e50404. ⟨10.3791/50404⟩. ⟨pasteur-01971522⟩

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