Applying Reverse Genetics to Study Measles Virus Interactions with the Host

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Vera-Peralta, Heidy | Najburg, Valérie | Combredet, Chantal | Douché, Thibaut | Giai Gianetto, Quentin | Matondo, Mariette | Tangy, Frédéric | Mura, Marie | Komarova, Anastassia

Edité par CCSD ; Springer -

International audience. The study of virus-host interactions is essential to achieve a comprehensive understanding of the viral replication process. The commonly used methods are yeast two-hybrid approach and transient expression of a single tagged viral protein in host cells followed by affinity purification of interacting cellular proteins and mass spectrometry analysis (AP-MS). However, by these approaches, virus-host protein-protein interactions are detected in the absence of a real infection, not always correctly compartmentalized, and for the yeast two-hybrid approach performed in a heterologous system. Thus, some of the detected protein-protein interactions may be artificial. Here we describe a new strategy based on recombinant viruses expressing tagged viral proteins to capture both direct and indirect protein partners during the infection (AP-MS in viral context). This way, virus-host protein-protein interacting co-complexes can be purified directly from infected cells for further characterization.

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