PT  - JOURNAL ARTICLE
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 Remy
AU  - Stoffel Michael
AU  - Heller Manfred
AU  - Dijkman Ronald
AU  - Thiel Volker
TI  - Determination of host cell proteins constituting the molecular microenvironment of coronavirus replicase complexes by proximity-labeling
AID  - 10.1101/417907
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 417907
4099  - http://biorxiv.org/content/early/2018/09/14/417907.short
4100  - http://biorxiv.org/content/early/2018/09/14/417907.full
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.