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Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay
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The small GTPase Rho5-Yet another player in yeast glucose signaling.

Franziska Schweitzer1, Linnet Bischof1, Stefan Walter2

  • 1Department of Biology/Chemistry, Division of Genetics, University of Osnabrück, Barbarastrasse, Osnabrück, Germany.

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Summary

The small GTPase Rho5 regulates yeast stress responses and carbohydrate metabolism. Rho5 deletion impacts cell wall and mitochondrial composition, affecting Ras/cAMP signaling.

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Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • Cellular Stress Response

Background:

  • The small GTPase Rho5 is implicated in Baker's yeast responses to cell wall stress, osmotic stress, and reactive oxygen species.
  • Rho5 and its GDP/GTP exchange factor translocate to mitochondria during stress and glucose starvation.

Purpose of the Study:

  • To investigate the role of Rho5 in carbohydrate metabolism and signaling pathways in Baker's yeast.
  • To elucidate the relationship between Rho5, glucose signaling, and stress response.

Main Methods:

  • Transcriptomic and proteomic analyses of rho5 deletion mutants.
  • Epistasis analysis with mutants in major yeast glucose signaling pathways.
  • Measurement of protein kinase A activity, glycogen, and trehalose levels.

Main Results:

  • Rho5 deletion affects carbohydrate metabolism at transcriptomic and proteomic levels.
  • Rho5 influences cell wall and mitochondrial composition.
  • Rho5 acts upstream of Ras2 GTPase in cAMP-mediated protein kinase A signaling, stimulating Ras/cAMP signaling.

Conclusions:

  • Rho5 plays a crucial role in regulating carbohydrate metabolism and stress responses in yeast.
  • Rho5 positively regulates the Ras/cAMP signaling pathway, impacting cellular homeostasis.