A study of "Super-resolution proximity labeling reveals antiviral protein network and its structural changes against SARS-CoV-2 viral proteins". Lee et al.
Description
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 “plug and playable” PL enzyme, TurboID-GBP (GFP binding protein) and we apply it for interactome mapping of SARS-CoV-2 ORF3a and M, which generates highly perturbed 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 obtains using the SR-PL method are presented in https://sarscov2.spatiomics.org. We hope that our method will contribute to revealing virus–host interactions of other viruses in an efficient manner.
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To determine the localization of SARS-CoV-2 viral protein, confocal microscopic imaging, EM (electron microscopic) imaging, and CLEM (correlative light and electron microscopic) imaging were conducted. LC-MS/MS was performed to analyze the interactome of SARS-CoV-2 viral protein. Detailed information is shown at attached files.
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Funding
National Research Foundation of Korea
NRF-2020K1A3A1A47110634, NRF-2021K2A9A1A01102293, NRF-2022R1A2B5B03001658, NRF-2022M3H9A2096199, 2022M3E5E8081185 to H.-W.R.; NRF-2022M3A9I2082294, NRF-2021R1A2C2009336 to J.-S.K.
Korea Health Industry Development Institute
Samsung Science and Technology Foundation (South Korea)
SSTF-BA2201-08
Korea Brain Research Institute
23-BR-01-03
Samsung (South Korea)
IO220819-02121-01