Genetic profiling of protein burden and nuclear export overload

Elife. 2020 Nov 4:9:e54080. doi: 10.7554/eLife.54080.

Abstract

Overproduction (op) of proteins triggers cellular defects. One of the consequences of overproduction is the protein burden/cost, which is produced by an overloading of the protein synthesis process. However, the physiology of cells under a protein burden is not well characterized. We performed genetic profiling of protein burden by systematic analysis of genetic interactions between GFP-op, surveying both deletion and temperature-sensitive mutants in budding yeast. We also performed genetic profiling in cells with overproduction of triple-GFP (tGFP), and the nuclear export signal-containing tGFP (NES-tGFP). The mutants specifically interacted with GFP-op were suggestive of unexpected connections between actin-related processes like polarization and the protein burden, which was supported by morphological analysis. The tGFP-op interactions suggested that this protein probe overloads the proteasome, whereas those that interacted with NES-tGFP involved genes encoding components of the nuclear export process, providing a resource for further analysis of the protein burden and nuclear export overload.

Keywords: S. cerevisiae; genetic profiling; genetics; genomics; nuclear export; overexpression; protein burden.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Active Transport, Cell Nucleus / genetics*
  • Cell Nucleus / metabolism
  • Genetic Profile
  • Genomics
  • Green Fluorescent Proteins
  • Mutation
  • Nuclear Export Signals / genetics*
  • Proteasome Endopeptidase Complex*
  • Protein Biosynthesis / genetics
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*

Substances

  • Nuclear Export Signals
  • Saccharomyces cerevisiae Proteins
  • Green Fluorescent Proteins
  • Proteasome Endopeptidase Complex