Research Paper Volume 8, Issue 12 pp 3255—3271

Slm35 links mitochondrial stress response and longevity through TOR signaling pathway

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Figure 1. SLM35 genetically interacts with TOR1 during stress conditions. (A) A strain lacking SLM35 shows resistance to stress by hydrogen peroxide. Log phase (upper panel) or stationary phase (lower panel) cultures of a wild-type (WT) and a Δslm35 with or without a plasmid with the SLM35 gene were treated with hydrogen peroxide (H2O2) for two hours, before serial ten-fold dilutions were dropped on rich media with glucose (YPD) or glycerol (YPG) as carbon sources. (B, C) Strains lacking SLM35slm35), components of the TOR/RAS pathways (Δtor1, Δsch9, Δras2 and Δrim15), or a combination of both, were subjected to (B) oxidative stress or to (C) heat shock as described in Materials and Methods. Serial 1:10 dilutions were dropped on rich media with glucose (YPD). (D) Total RNA from exponentially grown wild-type (WT), Δtor1, Δsch9, Δras2 and Δrim15 strains were purified. Equal amounts of RNA were used for retro-transcription reactions and resulting cDNAs were probed with SLM35 and ACT1 specific primers for the indicated cycles. Reactions were visualized by denaturing electrophoresis.