Sequestration of essential proteins causes prion associated toxicity in yeast

Mol Microbiol. 2009 Sep;73(6):1101-14. doi: 10.1111/j.1365-2958.2009.06836.x. Epub 2009 Aug 11.

Abstract

Prions are infectious, aggregated proteins that cause diseases in mammals but are not normally toxic in fungi. Excess Sup35p, an essential yeast protein that can exist as the [PSI(+)] prion, inhibits growth of [PSI(+)] but not [psi(-)] cells. This toxicity is rescued by expressing the Sup35Cp domain of Sup35p, which is sufficient for cell viability but not prion propagation. We now show that rescue requires Sup35Cp levels to be proportional to Sup35p overexpression. Overexpression of Sup35p appeared to cause pre-existing [PSI(+)] aggregates to coalesce into larger aggregates, but these were not toxic per se because they formed even when Sup35Cp rescued growth. Overexpression of Sup45p, but not other tested essential Sup35p binding partners, caused rescue. Sup45-GFPp formed puncta that colocalized with large [PSI(+)] Sup35-RFPp aggregates in cells overexpressing Sup35p, and the frequency of the Sup45-GFPp puncta was reduced by rescuing levels of Sup35Cp. In contrast, [PSI(+)] toxicity caused by a high excess of the Sup35p prion domain (Sup35NMp) was rescued by a single copy of Sup35Cp, was not rescued by Sup45p overexpression and was not associated with the appearance of Sup45-GFPp puncta. This suggests [PSI(+)] toxicity caused by excess Sup35p verses Sup35NMp is, respectively, through sequestration/inactivation of Sup45p verses Sup35p.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Gene Dosage
  • Microbial Viability
  • Peptide Termination Factors / genetics
  • Peptide Termination Factors / metabolism*
  • Prions / antagonists & inhibitors*
  • Prions / toxicity*
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Sequence Deletion

Substances

  • Peptide Termination Factors
  • Prions
  • SUP35 protein, S cerevisiae
  • SUP45 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins