Fig. 4.

AZC stress causes protein aggregation. (A) AZC treatment results in similar patterns of protein aggregates in wild-type and tsa1-mutant strains. Protein aggregates were isolated from wild-type and tsa1-mutant strains following 2 h of AZC treatment (5 mM) and were analysed by SDS-PAGE and silver staining. A, AZC treatment. (B) Hsp104–GFP forms multiple foci following AZC stress, and conversion to [rho⁰] does not affect the foci formation. The formation of Hsp104 foci following AZC treatment requires active protein synthesis because it is prevented by growth with cycloheximide (10 µg/ml, CHX). (C) The generation of ROS following AZC treatment requires active protein synthesis. Strains were grown to exponential phase in SD medium before treatment with CHX (200 µg/ml) or CHX plus AZC (5 mM) for 6 h. (D) Canavanine treatment results in ROS generation, which is abrogated in [rho⁰] cells. The indicated strains were grown to mid-exponential phase in SD medium, before treatment with canavanine (5 mM) for 6 h. (E) The formation of Hsp104 foci following canavanine treatment requires active protein synthesis.