Abstract
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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Acknowledgments
The authors would like to thank all past and present members of the Innovation Lab: Vaccines and the RNA and immunity laboratory. This work was supported by Institut Pasteur grants and BECAL Paraguay PhD grant to HV. Figures 2 and 3 were created with BioRender.com.
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The authors declare no conflicts of interest.
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Vera-Peralta, H. et al. (2024). Applying Reverse Genetics to Study Measles Virus Interactions with the Host. In: Ma, D.Z., Pfaller, C.K. (eds) Measles and Related Morbilliviruses. Methods in Molecular Biology, vol 2808. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3870-5_7
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DOI: https://doi.org/10.1007/978-1-0716-3870-5_7
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