title: Oxidative stress conditions increase the frequency of de novo formation of the yeast [PSI+] prion.
abstract: Prions are self-perpetuating amyloid protein aggregates which underlie various neurodegenerative diseases in mammals and heritable traits in yeast. The molecular basis of how yeast and mammalian prions form spontaneously into infectious amyloid-like structures is poorly understood. We have explored the hypothesis that oxidative stress is a general trigger for prion formation using the yeast [PSI(+)] prion, which is the altered conformation of the Sup35 translation termination factor. We show that the frequency of [PSI(+)] prion formation is elevated under conditions of oxidative stress and in mutants lacking key antioxidants. We detect increased oxidation of Sup35 methionine residues in antioxidant mutants and show that overexpression of methionine sulphoxide reductase abrogates both the oxidation of Sup35 and its conversion to the [PSI(+)] prion. [PSI(+)] prion formation is particularly elevated in a mutant lacking the Sod1 Cu,Zn-superoxide dismutase. We have used fluorescence microscopy to show that the de novo appearance of [PSI(+)] is both rapid and increased in frequency in this mutant. Finally, electron microscopy analysis of native Sup35 reveals that similar fibrillar structures are formed in both the wild-type and antioxidant mutants. Together, our data indicate that oxidative stress is a general trigger of [PSI(+) formation, which can be alleviated by antioxidant defenses.
authors.affiliation: Faculty of Life Sciences, University of Manchester, The Michael Smith Building, Oxford Road, Manchester, M13 9PT, UK.
authors.forename: Victoria A
authors.initials: VA
authors.lastname: Doronina
authors.forename: Gemma L
authors.initials: GL
authors.lastname: Staniforth
authors.forename: Shaun H
authors.initials: SH
authors.lastname: Speldewinde
authors.forename: Mick F
authors.initials: MF
authors.lastname: Tuite
authors.forename: Chris M
authors.initials: CM
authors.lastname: Grant
mesh_terms: D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D009154:Mutation; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010454:Peptide Termination Factors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000235:genetics
chemical_list: D010454:Peptide Termination Factors; C068068:SUP35 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D013482:Superoxide Dismutase