Super-resolution proximity labeling reveals anti-viral protein network and its structural changes against SARS-CoV-2 viral proteins
Year: 2023
Type: article
Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replicates in human cells by interacting with host factors following infection. To understand the virus and host interactome proximity, we introduce a super-resolution proximity labeling (SR-PL) method with a "plug-and-playable" PL enzyme, TurboID-GBP (GFP-binding nanobody protein), and we apply it for interactome mapping of SARS-CoV-2 ORF3a and membrane protein (M), which generates highly perturbed endoplasmic reticulum (ER) structures. Through SR-PL analysis of the biotinylated interactome, 224 and 272 peptides are robustly identified as ORF3a and M interactomes, respectively. Within the ORF3a interactome, RNF5 co-localizes with ORF3a and generates ubiquitin modifications of ORF3a that can be involved in protein degradation. We also observe that the SARS-CoV-2 infection rate is efficiently reduced by the overexpression of RNF5 in host cells. The interactome data obtained using the SR-PL method are presented at https://sarscov2.spatiomics.org. We hope that our method will contribute to revealing virus-host interactions of other viruses in an efficient manner.
Source: Cell Reports
Language: English
Topic: Biotin and Related Studies
Subfield: Cell Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $5,460
Funders: Korea Brain Research Institute, National Research Foundation, Seoul National University, Ministry of Science, ICT and Future Planning, Ministry of Health and Welfare, Korea Health Industry Development Institute, National Research Foundation of Korea, Vetenskapsrådet, Institute for Basic Science, Ministry of Science and ICT, South Korea, Samsung Science and Technology Foundation, Samsung
Awards: G1492580079, G1551050494, G1581705473, G1622723095, G165479020, G1802548948, G1883476937, G2022770724, Identification of immunological regulatory host factors for SARS-CoV-2 infection by proximity labeling analysis in the virus-infected cells, G2214948142, G2304812787, G2337071106, G2402660949, G2507944923, G2643796712, G3264138898, G3467338285, G3805646114, G3969051666, G4212643931, G5201466859, G5598398694, G6833455386, G7750438674, G8023915401, G8099688578, G8661181119, G8739444332
OpenAlex ID: W4385064043