A comprehensive library of fluorescent constructs of SARS‐CoV‐2 proteins and their initial characterisation in different cell types. (10th May 2021)

• Record Type: Journal Article
• Authors: Miserey‐Lenkei, Stéphanie; Trajkovic, Katarina; D'Ambrosio, Juan Martín; Patel, Amanda J; Čopič, Alenka; Mathur, Pallavi; Schauer, Kristine; Goud, Bruno; Albanèse, Véronique; Gautier, Romain; Subra, Melody; Kovacs, David; Barelli, Hélène; Antonny, Bruno
• Is Part Of: Biology of the cell. Volume 113:Number 7(2021)
• Page Start: 311
• Page End: 328
• Publication Date: 2021-05-10
• Subjects: Intracellular compartmentalisation -- Light microscopy -- Membranes -- Molecular interactions -- Viruses; Cytology -- Periodicals; Electron microscopy -- Periodicals; 571.6
• DOI: 10.1111/boc.202000158
• Languages: English
• ISSNs: 0248-4900

Abstract

Abstract : Background Information: Comprehensive libraries of plasmids for SARS‐CoV‐2 proteins with various tags ( e.g ., Strep, HA, Turbo) are now available. They enable the identification of numerous potential protein–protein interactions between the SARS‐CoV‐2 virus and host proteins. Results: We present here a large library of SARS CoV‐2 protein constructs fused with green and red fluorescent proteins and their initial characterisation in various human cell lines including lung epithelial cell models (A549, BEAS‐2B), as well as in budding yeast. The localisation of a few SARS‐CoV‐2 proteins matches their proposed interactions with host proteins. These include the localisation of Nsp13 to the centrosome, Orf3a to late endosomes and Orf9b to mitochondria. Conclusions and Significance: This library should facilitate further cellular investigations, notably by imaging techniques. Abstract : Research article : Comprehensive proteomic studies have suggested numerous interactions between SARS‐CoV‐2 and host cellular proteins. We prepared SARS‐CoV‐2 GFP or mCherry fusion constructs and analysed their cellular localisation. The localisation a few SARS‐CoV‐2 fluorescent proteins matches their proposed molecular interactions. These include Nsp13 at the centrosome, Orf3a on late endosomes and Orf9b at mitochondria.