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The Goldilocks effect of respiration on canavanine tolerance in Saccharomyces cerevisiae

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Abstract

When glucose is available, Saccharomyces cerevisiae prefers fermentation to respiration. In fact, it can live without respiration at all. Here, we study the role of respiration in stress tolerance in yeast. We found that colony growth of respiratory-deficient yeast (petite) is greatly inhibited by canavanine, the toxic analog of arginine that causes proteotoxic stress. We found lower amounts of the amino acids involved in arginine biosynthesis in petites compared with WT. This finding may be explained by the fact that petite cells exposed to canavanine show reduction in the efficiency of targeting of proteins required for arginine biosynthesis. The retrograde (RTG) pathway signals mitochondrial stress. It positively controls production of arginine precursors. We show that canavanine abrogates RTG signaling especially in petite cells, and mutants in the RTG pathway are extremely sensitive to canavanine. We suggest that petite cells are naturally ineffective in production of some amino acids; combination of this fact with the effect of canavanine on the RTG pathway is the simplest explanation why petite cells are inhibited by canavanine. Surprisingly, we found that canavanine greatly inhibits colony formation when WT cells are forced to respire. Our research proposes a novel connection between respiration and proteotoxic stress.

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Acknowledgements

We thank the Schuldiner lab for technical support and most valuable input. This work was supported by RCDA Grant of ICRF to SC.

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Authors and Affiliations

  1. Department of Plant Pathology and Microbiology, Robert H. Smith Faculty of Agriculture, Food and Environment, Hebrew University, 76100, Rehovot, Israel

    Marina Druseikis & Shay Covo

  2. Interdepartmental Equipment Unit, Robert H. Smith Faculty of Agriculture, Food and Environment, Hebrew University, 76100, Rehovot, Israel

    Julius Ben-Ari

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  1. Marina Druseikis
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Correspondence to Shay Covo.

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Communicated by M. Kupiec.

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Arginine biosynthesis pathway.

Original representation based on arginine biosynthesis pathway from SGD database (yeastgenome.org). Red denotes location in mitochondria; blue denotes location in cytosol (TIFF 1540 kb)

294_2019_974_MOESM2_ESM.tiff

can1Δis sensitive to growth on a non-fermentable carbon source with canavanine. A can1Δ mutant was pronged onto glucose (-) arginine with 20 μg/ml canavanine and lactic acid (-) arginine with 20 μg/ml canavanine. Growth after one week (TIFF 104 kb)

Arginine addition somewhat rescues petites.

Strains were grown on 2 µg/ml canavanine glucose (-) arginine and glucose with increasing amounts of arginine. Growth after 10 days (TIFF 739 kb)

Supplementary material 4 (XLSX 24 kb)

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Druseikis, M., Ben-Ari, J. & Covo, S. The Goldilocks effect of respiration on canavanine tolerance in Saccharomyces cerevisiae. Curr Genet 65, 1199–1215 (2019). https://doi.org/10.1007/s00294-019-00974-y

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