Ribopuromycylation in Coronavirus-Infected Cells

Methods Mol Biol. 2020:2203:231-238. doi: 10.1007/978-1-0716-0900-2_17.

Abstract

Ribopuromycylation enables the visualization and quantitation of translation on a cellular level by immunofluorescence or in total using standard western blotting. This technique uses ribosome catalyzed puromycylation of nascent chains followed by immobilization on the ribosome by antibiotic chain elongation inhibitor emetine. Detection of puromycylated ribosome-bound nascent chains can then be achieved using a puromycin-specific antibody.

Keywords: Emetine; Puromycin; Puromycylation; Ribopuromycylation; Translation.

MeSH terms

  • Coronavirus / genetics*
  • Coronavirus Infections
  • Fluorescent Antibody Technique
  • Host-Pathogen Interactions
  • Humans
  • Protein Biosynthesis
  • Puromycin / pharmacology*
  • Ribosomes / drug effects
  • Ribosomes / metabolism

Substances

  • Puromycin