Fig. 2: Impact of cell integrity signaling on phosphorylation of T492.
From: Mechanosensor-mediated Hsp70 phosphorylation orchestrates the landscape of the heat shock response
a The yeast cell wall integrity pathway. Created in BioRender. Truman, A. (2025) https://BioRender.com/ibts1wxb Impact of Mid2 and Wsc1 mechanosensors on T492 phosphorylation. Lysates from indicated cells were analyzed by Western Blotting using antisera to either FLAG or phospho-T492. c T492 phosphorylation is triggered by membrane stretch. Lysate from FLAG-WT or T492A cells either untreated or treated with chlorpromazine (CPZ) were analyzed by Western Blotting using antisera to either FLAG or phospho-T492. d T492 phosphorylation is dependent on Pkc1. FLAG-Ssa1 (WT or with genomically AID-tagged Pkc1) cells were treated with auxin for the indicated times followed by 30-minute incubation at 39 °C. Lysates were analyzed by Western Blotting using antisera to either FLAG or phospho-T492. e T492 phosphorylation is independent of Bck1, Mkk1, Mkk2 and Mpk1. Lysate from indicated knockout strains were analyzed by Western Blotting using antisera to either FLAG or phospho-T492. f Pkc1 directly phosphorylates Ssa1 T492. An in vitro kinase assay was performed with HA-Pkc1 purified from heat-shocked yeast and recombinant Ssa1. The reaction mixture was analyzed by Western Blotting using antisera to HA, phospho-T492 and HIS epitope. Each experiment involving western blotting was repeated three times with similar results. Source data are provided as a Source Data file.
