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

Biol Cell. 2021 Jul;113(7):311-328. doi: 10.1111/boc.202000158. Epub 2021 May 10.

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.

Keywords: Intracellular compartmentalisation; Light microscopy; Membranes; Molecular interactions; Viruses.

MeSH terms

  • A549 Cells
  • COVID-19 / virology*
  • Cell Line
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Host Microbial Interactions / physiology
  • Humans
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Microscopy, Fluorescence
  • Peptide Library*
  • Protein Interaction Domains and Motifs
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Red Fluorescent Protein
  • SARS-CoV-2 / genetics
  • SARS-CoV-2 / metabolism*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Time-Lapse Imaging
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*

Substances

  • Luminescent Proteins
  • Peptide Library
  • Recombinant Fusion Proteins
  • Viral Proteins
  • Green Fluorescent Proteins