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

V'kovski, Philip; Gerber, M; Kelly, Jenna Nicole; Pfänder, Stephanie; Ebert, Nadine; Braga, Sophie Marie-Pierre; Simillion, Cedric André Marie; Portmann, Jasmine; Stalder, Hanspeter; Gaschen, Véronique; Bruggmann, Rémy; Stoffel, Michael Hubert; Heller, Manfred; Dijkman, Ronald; Thiel, Volker Earl (14 September 2018). Determination of host cell proteins constituting the molecular microenvironment of coronavirus replicase complexes by proximity-labeling (bioRxiv). Cold Spring Harbor Laboratory 10.1101/417907

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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.

Item Type:

Working Paper

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR)
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH) > Experimental Clinical Research
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Virology and Immunology
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH) > Veterinary Anatomy
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)
09 Interdisciplinary Units > Microscopy Imaging Center MIC
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > DCR Services > Genomics

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

V'kovski, Philip; Kelly, Jenna Nicole; Pfänder, Stephanie; Ebert, Nadine; Braga, Sophie Marie-Pierre; Simillion, Cedric André Marie; Portmann, Jasmine; Stalder, Hanspeter; Gaschen, Véronique; Bruggmann, Rémy; Stoffel, Michael Hubert; Heller, Manfred; Dijkman, Ronald and Thiel, Volker Earl

Subjects:

600 Technology > 630 Agriculture
600 Technology > 610 Medicine & health
500 Science > 570 Life sciences; biology

Series:

bioRxiv

Publisher:

Cold Spring Harbor Laboratory

Language:

English

Submitter:

Philip V'kovski

Date Deposited:

28 Nov 2018 09:06

Last Modified:

07 Jun 2019 14:40

Publisher DOI:

10.1101/417907

Additional Information:

unpublished preprint

BORIS DOI:

10.7892/boris.121510

URI:

https://boris.unibe.ch/id/eprint/121510

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