TY - JOUR TI - Determination of host proteins composing the microenvironment of coronavirus replicase complexes by proximity-labeling AU - V'kovski, Philip AU - Gerber, Markus AU - Kelly, Jenna AU - Pfaender, Stephanie AU - Ebert, Nadine AU - Braga Lagache, Sophie AU - Simillion, Cedric AU - Portmann, Jasmine AU - Stalder, Hanspeter AU - Gaschen, Véronique AU - Bruggmann, Rémy AU - Stoffel, Michael H AU - Heller, Manfred AU - Dijkman, Ronald AU - Thiel, Volker A2 - Kirkegaard, Karla A2 - Dikic, Ivan VL - 8 PY - 2019 DA - 2019/01/11 SP - e42037 C1 - eLife 2019;8:e42037 DO - 10.7554/eLife.42037 UR - https://doi.org/10.7554/eLife.42037 AB - Positive-sense RNA viruses hijack intracellular membranes that provide niches for viral RNA synthesis and a platform for interactions with host proteins. However, little is known about host factors at the interface between replicase complexes and the host cytoplasm. We engineered a biotin ligase into a coronaviral replication/transcription complex (RTC) and identified >500 host proteins constituting the RTC microenvironment. siRNA-silencing of each RTC-proximal host factor demonstrated importance of vesicular trafficking pathways, ubiquitin-dependent and autophagy-related processes, and translation initiation factors. Notably, detection of translation initiation factors at the RTC was instrumental to visualize and demonstrate active translation proximal to replication complexes of several coronaviruses. Collectively, we establish a spatial link between viral RNA synthesis and diverse host factors of unprecedented breadth. Our data may serve as a paradigm for other positive-strand RNA viruses and provide a starting point for a comprehensive analysis of critical virus-host interactions that represent targets for therapeutic intervention. KW - coronavirus KW - replicase complex KW - proximity labeling KW - virus-host interaction KW - translation KW - vesicular transport JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -