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Peroxiredoxin promotes longevity and H2O2-resistance in yeast through redox-modulation of protein kinase A

ABSTRACT: Peroxiredoxins are modulators of aging in yeast and multicellular organisms. The mechanisms by which peroxiredoxins stimulate H2O2 resistance and slow down aging are, however, unclear. Here we show that the yeast peroxiredoxin Tsa1 boosts H2O2 resistance and prevents aging independent of cellular H2O2 levels, but in a manner dependent on H2O2 signaling. More specifically, we pin-point a role of Tsa1 in repressing nutrient signaling via protein kinase A (PKA) that governs both H2O2 resistance and longevity. Tsa1 controls PKA activity at the level of the catalytic subunits and Tpk1 is redox-modified by Tsa1 on a conserved cysteine residue (Cys243) in Tpk1 upon H2O2 addition boosting cellular H2O2 resistance. Tpk1 redox modification dephosphorylates a conserved threonine 241 in the activation loop reducing enzyme activity. We discuss these results in the context of an integrative view on aging where nutrient signaling pathways constitute hubs integrating information from multiple aging-related conduits.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Wolfgang Reiter

LAB HEAD: Mikael Molin

PROVIDER: PXD012617 | Pride | 2020-07-27

REPOSITORIES: pride

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			Action	DRS
	20180918_Lumos_RSLC6_Reiter_Ammerer_MFPL_onbead_Tpk1_1_1.raw	Raw
	20180918_Lumos_RSLC6_Reiter_Ammerer_MFPL_onbead_Tpk1_1_2.raw	Raw
	20180918_Lumos_RSLC6_Reiter_Ammerer_MFPL_onbead_Tpk1_1_3.raw	Raw
	20180918_Lumos_RSLC6_Reiter_Ammerer_MFPL_onbead_Tpk1_2_1.raw	Raw
	20180918_Lumos_RSLC6_Reiter_Ammerer_MFPL_onbead_Tpk1_2_2.raw	Raw

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Project description:Cells must adjust their gene expression in order to compete in a constantly changing environment. Two alternative strategies could in principle ensure optimal coordination of gene expression with physiological requirement. First, the internal physiological state itself could feedback to regulated gene expression. Second, the expected physiological state could be inferred from the external environment, using evolutionary-tuned signaling pathways. Coordination of ribosomal biogenesis with the requirement for protein synthesis appears to be particularly important, since cells devote a large fraction of their biosynthetic capacity for ribosomal biogenesis. To define the relative importance of internal vs. external sensing to the regulation of ribosomal biogenesis gene expression, we subjected S. cerevisiae cells to conditions which decoupled the actual vs. environmentally-expected growth rate. Gene expression followed the environmental signal according to the expected, but not the actual, growth rate. Simultaneous monitoring of gene expression and growth rate in chemostat-grown cultures further confirmed that ribosome biogenesis genes responded rapidly to changes in the environments but were oblivious to longer-term changes in growth rate. Our results suggest that the capacity to anticipate and prepare for environmental changes presented a major selection force during yeast evolution. Keywords: Saccharomyces_Cerevisiae, Stress response, ADH1 deletion, time courses, chemostat Experiment 1: ADH1 deletion cells, which grow faster on glycerol rather then glucose medium, were grown in both glucose and glycerol medium. The cells expression on glycerol is compared to their expression glucose (5 microarrays). Experiment 2: Time course of ADH1 deletion cells upon growth on glycerol and on glucose (7 microarrays). Experiment 3: Response of steady state grown cells to environmental perturbations. Cells were grown in glucose or histidine limited chemostats and reached steady state. The cells were then subjected to perturbations such as DTT, heat shock, NaCl, Clotrimazole, H2O2, and addition of limiting factors (histidine and glucose, respectively). Overall we have examined 10 timecourses with 83 microarrays.

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Peroxiredoxin promotes longevity and H2O2-resistance in yeast through redox-modulation of protein kinase A

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