The yeast peroxiredoxin Tsa1 protects against protein-aggregate-induced oxidative stress

Peroxiredoxins are ubiquitous thiol-specific proteins that have multiple functions in stress protection, including protection against oxidative stress. Tsa1 is the major yeast peroxiredoxin and we show that it functions as a specific antioxidant to protect the cell against the oxidative stress caused by nascent-protein misfolding and aggregation. Yeast mutants lacking TSA1 are sensitive to misfolding caused by exposure to the proline analogue azetidine-2-carboxylic acid (AZC). AZC promotes protein aggregation, and its toxicity to a tsa1 mutant is caused by the production of reactive oxygen species (ROS). The generation of [rho 0] cells, which lack mitochondrial DNA, rescues the tsa1 mutant AZC sensitivity, indicating that mitochondria are the source of ROS. Inhibition of nascent-protein synthesis with cycloheximide prevents AZC-induced protein aggregation and abrogates ROS generation, confirming that the formation of aggregates causes ROS production. Protein aggregation is accompanied by mitochondrial fragmentation, and we show that Tsa1 localises to the sites of protein aggregation. Protein aggregates are formed adjacent to mitochondria, and our data indicate that active mitochondria generate ROS. These data indicate a new role for peroxiredoxins in protecting against ROS that are generated as a result of protein misfolding and aggregate formation.