Super-resolution proximity labeling reveals anti-viral protein network and its structural changes against SARS-CoV-2 viral proteins.

Lee Y, Jung M, Kim J, Charles A, Christ W, Kang J, Kang M, Kwak C, Klingström J, Smed-Sörensen A, Kim J, Mun JY, Rhee H

Cell Rep 42 (8) 112835 [2023-08-29; online 2023-07-20]

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 We hope that our method will contribute to revealing virus-host interactions of other viruses in an efficient manner.

Category: Proteins

Type: Journal article

PubMed 37478010

DOI 10.1016/j.celrep.2023.112835

Crossref 10.1016/j.celrep.2023.112835

pii: S2211-1247(23)00846-X

Publications 9.5.0