Proximity Labeling for the Identification of Coronavirus-Host Protein Interactions.

V'kovski, Philip; Steiner, Silvio; Thiel, Volker (2020). Proximity Labeling for the Identification of Coronavirus-Host Protein Interactions. Methods in molecular biology, 2203, pp. 187-204. Springer 10.1007/978-1-0716-0900-2_14

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Biotin-based proximity labeling circumvents major pitfalls of classical biochemical approaches to identify protein-protein interactions. It consists of enzyme-catalyzed biotin tags ubiquitously apposed on proteins located in close proximity of the labeling enzyme, followed by affinity purification and identification of biotinylated proteins by mass spectrometry. Here we outline the methods by which the molecular microenvironment of the coronavirus replicase/transcriptase complex (RTC), i.e., proteins located within a close perimeter of the RTC, can be determined by different proximity labeling approaches using BirAR118G (BioID), TurboID, and APEX2. These factors represent a molecular signature of coronavirus RTCs and likely contribute to the viral life cycle, thereby constituting attractive targets for the development of antiviral intervention strategies.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Virology and Immunology
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

V'kovski, Philip; Steiner, Silvio and Thiel, Volker Earl

Subjects:

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

ISSN:

1940-6029

Publisher:

Springer

Language:

English

Submitter:

Achim Braun Parham

Date Deposited:

08 Apr 2021 08:56

Last Modified:

08 Apr 2021 08:56

Publisher DOI:

10.1007/978-1-0716-0900-2_14

PubMed ID:

32833213

Uncontrolled Keywords:

Affinity purification Ascorbate peroxidase APEX2 BioID Biotin ligase Coronavirus Mass spectrometry Proteomics Proximity labeling Replicase microenvironment Replication transcription complex RTC TurboID

BORIS DOI:

10.7892/boris.152071

URI:

https://boris.unibe.ch/id/eprint/152071

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