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Genome-wide identification of genes involved in tolerance to various environmental stresses in Saccharomyces cerevisiae

100%

Auesukaree C., Damnernsawad A., Kruatrachue M., Pokethitiyook P., Boonchird C., Kaneko Y., Harashima S.

Journal of Applied Genetics

2009

vol. 50

issue 3

301-310

During fermentation, yeast cells are exposed to a number of stresses ? such as high alcohol concentration, high osmotic pressure, and temperature fluctuation ? so some overlap of mechanisms involved in the response to these stresses has been suggested. To identify the genes required for tolerance to alcohol (ethanol, methanol, and 1-propanol), heat, osmotic stress, and oxidative stress, we performed genome-wide screening by using 4828 yeast deletion mutants. Our screens identified 95, 54, 125, 178, 42, and 30 deletion mutants sensitive to ethanol, methanol, 1-propanol, heat, NaCl, and H2O2, respectively. These deleted genes were then classified based on their cellular functions, and cross-sensitivities between stresses were determined. A large number of genes involved in vacuolar H+-ATPase (V-ATPase) function, cytoskeleton biogenesis, and cell wall integrity, were required for tolerance to alcohol, suggesting their protective role against alcohol stress. Our results revealed a partial overlap between genes required for alcohol tolerance and those required for thermotolerance. Genes involved in cell wall integrity and the actin cytoskeleton are required for both alcohol tolerance and thermotolerance, whereas the RNA polymerase II mediator complex seems to be specific to heat tolerance. However, no significant overlap of genes required for osmotic stress and oxidative stress with those required for other stresses was observed. Interestingly, although mitochondrial function is likely involved in tolerance to several stresses, it was found to be less important for thermotolerance. The genes identified in this study should be helpful for future research into the molecular mechanisms of stress response.

Effects of in vitro conditions and in vivo thermal adaptation on viability of the earthworm (Eisenia fetida) coelomocytes

86%

Stankiewicz A., Plytycz B.

Folia Biologica

1998

vol. 46

183-188

Coelomocytes of E. fetida were extruded from earthworms immersed for 1 minute in fluid and treated either with an electric shock (5V) or 5% ethanol. The two methods were equally efficient with regard to the number and viability of coelomocytes retrieved and did not affect animal survival. Cell viability was not affected by short contact with the extrusion fluids of wide ranges of osmolarities (10-1600 mOsm) and pH (3-11). In contrast, extreme pH (especially 11) and osmolarities (especially 22 and 600 mOsm) impaired cell viability during a few hours of in vitro incubation. Cell survival was best in L-15 medium at the neutral pH and at the osmolarity characteristic for terrestrial animals (285-320 mOsm). Coelomocyte viability after 7 hr in vitro incubation at 0, 10, 22, and 37oC was always the lowest at 37oC. Cells extruded from animals maintained at room temperature were the best vital at 22oC. Viability of the cells transferred from 22oC to 10oC for one week only was best at 10oC and equally good at 0oC and 22oC. In vitro viability of cells retrieved from earthworms maintained at 10oC for 2 or 3 weeks was significantly higher at 10oC and 0oC than at 22oC. This indicates that a relatively long-lasting (more than one week) in vivo adaptation to cold is necessary to improve coelomocyte viability at low temperatures.

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1 Folia Biologica

1 Journal of Applied Genetics

1 2009

1 1998

1 Auesukaree C.

1 Boonchird C.

1 Damnernsawad A.

1 Harashima S.

1 Kaneko Y.

1 Kruatrachue M.

1 Plytycz B.

1 Pokethitiyook P.

1 Stankiewicz A.

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Genome-wide identification of genes involved in tolerance to various environmental stresses in Saccharomyces cerevisiae

Effects of in vitro conditions and in vivo thermal adaptation on viability of the earthworm (Eisenia fetida) coelomocytes