A comprehensive library of fluorescent constructs of SARS‐CoV‐2 proteins and their initial characterisation in different cell types

• Published in: Biology of the Cell Vol. 113; no. 7; pp. 311 - 328
• Main 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
• Format: Journal Article Web Resource
• Language: English
• Published: England John Wiley & Sons, Inc 01.07.2021; Wiley; John Wiley and Sons Inc
• Subjects: A549 Cells; Cell Line; Cellular Biology; COVID-19 - virology; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Host Microbial Interactions - physiology; Humans; Intracellular compartmentalisation; Life Sciences; Light microscopy; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Membranes; Microscopy, Fluorescence; Molecular interactions; Peptide Library; Protein Interaction Domains and Motifs; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Red Fluorescent Protein; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; SARS-CoV-2 - genetics; SARS-CoV-2 - metabolism; Time-Lapse Imaging; Viral Proteins - genetics; Viral Proteins - metabolism; Viruses; Intracellular compartmentalisation; Light microscopy; Viruses; Membranes; Molecular interactions
• ISSN: 0248-4900; 1768-322X
• DOI: 10.1111/boc.202000158

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. 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.