Determination of host proteins composing the microenvironment of coronavirus replicase complexes by proximity-labeling.

V'kovski, Philip; Gerber, Markus; Kelly, Jenna Nicole; Pfänder, Stephanie; Ebert, Nadine; Braga Lagache, Sophie; Simillion, Cedric; Portmann, Jasmine; Stalder, Hanspeter; Gaschen, Véronique; Bruggmann, Rémy; Stoffel, Michael Hubert; Heller, Manfred; Dijkman, Ronald; Thiel, Volker Earl (2019). Determination of host proteins composing the microenvironment of coronavirus replicase complexes by proximity-labeling. eLife, 8 eLife Sciences Publications 10.7554/eLife.42037

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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:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Research Foci > Host-Pathogen Interaction
04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases > Research
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)
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > DCR Services > Core Facility Massenspektrometrie- und Proteomics-Labor
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Unit Childrens Hospital > Protein- und Zellbiologie

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

V'kovski, Philip; Gerber, Markus Daniel; Kelly, Jenna Nicole; Pfänder, Stephanie; Ebert, Nadine; Portmann, Jasmine; Stalder, Hanspeter; Gaschen, Véronique; Bruggmann, Rémy; Stoffel, Michael Hubert; Heller, Manfred; Dijkman, Ronald and Thiel, Volker Earl

Subjects:

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

ISSN:

2050-084X

Publisher:

eLife Sciences Publications

Language:

English

Submitter:

Achim Braun Parham

Date Deposited:

04 Sep 2019 16:42

Last Modified:

04 Sep 2019 16:42

Publisher DOI:

10.7554/eLife.42037

PubMed ID:

30632963

Uncontrolled Keywords:

coronavirus human infectious disease microbiology mouse proximity labeling replicase complex translation vesicular transport virus virus-host interaction

BORIS DOI:

10.7892/boris.132988

URI:

https://boris.unibe.ch/id/eprint/132988

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