title: 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.
authors.affiliation: Unité de Génétique, Papillomavirus et Cancer Humain (GPCH), Institut Pasteur.
authors.forename: Mandy
authors.initials: M
authors.lastname: Muller
authors.forename: Patricia
authors.initials: P
authors.lastname: Cassonnet
authors.forename: Michel
authors.initials: M
authors.lastname: Favre
authors.forename: Yves
authors.initials: Y
authors.lastname: Jacob
authors.forename: Caroline
authors.initials: C
authors.lastname: Demeret
mesh_terms: D028023:Arecaceae / Q000201:enzymology; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D054884:Host-Pathogen Interactions; D006801:Humans; D008156:Luciferases / Q000737:chemistry / Q000378:metabolism; D060066:Protein Interaction Maps; D014162:Transfection / Q000379:methods; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics / Q000378:metabolism
chemical_list: D008156:Luciferases
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