Antiprion systems in yeast cooperate to cure or prevent the generation of nearly all [ PSI + ] and [URE3] prions.

[ PSI ⁺ ] and [URE3] are prions of Saccharomyces cerevisiae based on amyloids of Sup35p and Ure2p, respectively. In normal cells, antiprion systems block prion formation, cure many prions that arise, prevent infection by prions, and prevent toxicity of those prions that escape the other systems. The upf1Δ , ssz1Δ , and hsp104 ^T160M single mutants each develop [ PSI ⁺ ] at 10- to 15-fold, but the triple mutant spontaneously generates [ PSI ⁺ ] at up to ∼5,000-fold the wild-type rate. Most such [ PSI ⁺ ] variants are cured by restoration of any one of the three defective antiprion systems, defining a previously unknown type of [ PSI ⁺ ] variant and proving that these three antiprion systems act independently. Generation of [ PSI ⁺ ] variants stable in wild-type cells is also increased in upf1Δ ssz1Δ hsp104 ^T160M strains 25- to 500-fold. Btn2 and Cur1 each cure 90% of [URE3] prions generated in their absence, but we find that btn2Δ or cur1Δ diminishes the frequency of [ PSI ⁺ ] generation in an otherwise wild-type strain. Most [ PSI ⁺ ] isolates in a wild-type strain are destabilized on transfer to a btn2Δ or cur1Δ host. Single upf1Δ or hsp104 ^T160M mutants show the effects of btn2Δ or cur1Δ but not upf1Δ ssz1Δ hsp104 ^T160M or ssz1Δ hsp104 ^T160M strains. The disparate action of Btn2 on [URE3] and [ PSI ⁺ ] may be a result of [ PSI ⁺ ]'s generally higher seed number and lower amyloid structural stability compared with [URE3]. Thus, prion generation is not a rare event, but the escape of a nascent prion from the surveillance by the antiprion systems is indeed rare.