Determination of host cell proteins constituting the molecular microenvironment of coronavirus replicase complexes by proximity-labeling

V’kovski Philip; Gerber Markus; Kelly Jenna; Pfaender Stephanie; Ebert Nadine; Braga Lagache Sophie; Simillion Cedric; Portmann Jasmine; Stalder Hanspeter; Gaschen Véronique; Bruggmann Remy; Stoffel Michael; Heller Manfred; Dijkman Ronald; Thiel Volker

Consistently, cell-associated viral mRNA levels (Fig. 4d ) and viral titers ( The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/417907 doi: bioRxiv preprint coronaviruses, such as HCoV-229E or the highly pathogenic MERS-CoV (Fig. 6) . The HCoV-331 229E RTC, which was detected with an antiserum directed against nsp8, appeared as small and 332 dispersed perinuclear puncta during early infection and eventually converged into larger 333 perinuclear structures later in infection. Consistent with findings obtained for MHV, we 334 observed a striking co-localization of the HCoV-229E RTC with sites of active translation 335 during the early phase of the infection (Fig. 6, S4) . The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/417907 doi: bioRxiv preprint defined temporal roles during particular phases of the viral life cycle and proteins that did not 380 yet attract our attention in previous screens because of functional redundancies. We therefore 381 expect that this approach will find wide application in the field of virus-host interaction, target 382 identification for virus inhibition, and provides a starting point to reveal similarities and 383 differences between replication strategies of a broad range of viruses. 384