Fig. 5.

Tsa1 colocalises with sites of protein aggregation. (A) Tsa1 colocalises with Hsp104 following AZC stress. A wild-type strain coexpressing Tsa1–GFP and Hsp104–RFP was grown to mid-exponential phase before AZC (5 mM) treatment. (B) Confirmation of the colocalisation of Tsa1 and Hsp104. Images were acquired on a Deltavision restoration microscope, deconvolved and presented as a maximum-intensity projection from optical sections. (C) The formation of Tsa1–GFP foci in response to AZC, but not H₂O₂, requires nascent-protein synthesis because it is abrogated by cycloheximide (CHX) treatment. (D) Cys⁴⁸ is not required for the localisation of Tsa1 to insoluble protein aggregates. Insoluble protein aggregates were prepared as in Fig. 4A and Tsa1 was detected by western blot. A, AZC treatment.