Hyperosmotic stress induces metacaspase - and mitochondria - dependent apoptosis in Saccharomyces cerevisiae

Prova tipográfica (In Press)During the last years, several reports described an apoptosis-like programmed cell death process in yeast in response to different environmental aggressions. Here, evidence is presented that hyperosmotic stress induces in Saccharomyces cerevisiae a cell death process accompanied by morphological and biochemical indicators of apoptotic programmed cell death, namely chromatin condensation along the nuclear envelope, mitochondrial swelling and reduction of cristae number, production of reactive oxygen species and DNA strand breaks, with maintenance of plasma membrane integrity. Disruption of AIF1 had no effect on cell survival, but lack of Yca1p drastically reduced metacaspase activation and decreased cell death indicating that this death process was associated to activation of this protease. Supporting the involvement of mitochondria and cytochrome c in caspase activation, the mutant strains cyc1Δ cyc7Δ and cyc3Δ, both lacking mature cytochrome c, displayed a decrease in caspase activation associated to increased cell survival when exposed to hyperosmotic stress. These findings indicate that hyperosmotic stress triggers S. cerevisiae into an apoptosis-like programmed cell death that is mediated by a caspase-dependent mitochondrial pathway partially dependent on cytochrome c

Quantitative Proteomics of Yeast Post-Golgi Vesicles Reveals a Discriminating Role for Sro7p in Protein Secretion

We here report the first comparative proteomics of purified yeast post-Golgi vesicles (PGVs). Vesicle samples isolated from PGV-accumulating sec6-4 mutants were treated with isobaric tags (iTRAQ) for subsequent quantitative tandem mass spectrometric analysis of protein content. After background subtraction, a total of 66 vesicle-associated proteins were identified, including known or assumed vesicle residents as well as a fraction not previously known to be PGV associated. Vesicles isolated from cells lacking the polarity protein Sro7p contained essentially the same catalogue of proteins but showed a reduced content of a subset of cargo proteins, in agreement with a previously shown selective role for Sro7p in cargo sorting

Lack of Trehalose Accelerates H2O2-Induced Candida albicans Apoptosis through Regulating Ca2+ Signaling Pathway and Caspase Activity

Trehalose is a non-reducing disaccharide and can be accumulated in response to heat or oxidative stresses in Candida albicans. Here we showed that a C. albicans tps1Δ mutant, which is deficient in trehalose synthesis, exhibited increased apoptosis rate upon H2O2 treatment together with an increase of intracellular Ca2+ level and caspase activity. When the intracellular Ca2+ level was stimulated by adding CaCl2 or A23187, both the apoptosis rate and caspase activity were increased. In contrast, the presence of two calcium chelators, EGTA and BAPTA, could attenuate these effects. Moreover, we investigated the role of Ca2+ pathway in C. albicans apoptosis and found that both calcineurin and the calcineurin-dependent transcription factor, Crz1p, mutants showed decreased apoptosis and caspase activity upon H2O2 treatment compared to the wild-type cells. Expression of CaMCA1, the only gene found encoding a C. albicans metacaspase, in calcineurin-deleted or Crz1p-deleted cells restored the cell sensitivity to H2O2. Our results suggest that Ca2+ and its downstream calcineurin/Crz1p/CaMCA1 pathway are involved in H2O2 -induced C. albicans apoptosis. Inhibition of this pathway might be the mechanism for the protective role of trehalose in C. albicans

A Newly Identified Essential Complex, Dre2-Tah18, Controls Mitochondria Integrity and Cell Death after Oxidative Stress in Yeast

A mutated allele of the essential gene TAH18 was previously identified in our laboratory in a genetic screen for new proteins interacting with the DNA polymerase delta in yeast [1]. The present work shows that Tah18 plays a role in response to oxidative stress. After exposure to lethal doses of H2O2, GFP-Tah18 relocalizes to the mitochondria and controls mitochondria integrity and cell death. Dre2, an essential Fe/S cluster protein and homologue of human anti-apoptotic Ciapin1, was identified as a molecular partner of Tah18 in the absence of stress. Moreover, Ciapin1 is able to replace yeast Dre2 in vivo and physically interacts with Tah18. Our results are in favour of an oxidative stress-induced cell death in yeast that involves mitochondria and is controlled by the newly identified Dre2-Tah18 complex

Regulation of the Na+/K+-ATPase Ena1 Expression by Calcineurin/Crz1 under High pH Stress: A Quantitative Study

[EN] Regulated expression of the Ena1 Na+-ATPase is a crucial event for adaptation to high salt and/or alkaline pH stress in the budding yeast Saccharomyces cerevisiae. ENA1 expression is under the control of diverse signaling pathways, including that mediated by the calcium-regulatable protein phosphatase calcineurin and its downstream transcription factor Crz1. We present here a quantitative study of the expression of Ena1 in response to alkalinization of the environment and we analyze the contribution of Crz1 to this response. Experimental data and mathematical models substantiate the existence of two stress-responsive Crz1-binding sites in the ENA1 promoter and estimate that the contribution of Crz1 to the early response of the ENA1 promoter is about 60%. The models suggest the existence of a second input with similar kinetics, which would be likely mediated by high pH-induced activation of the Snf1 kinase.This work was supported by grants BFU2011-30197-C3-01, BFU2014-54591-C2-1-P and EUI2009-04147 (SysMo2) to JA. (Ministry of Industry and Competitivity, Spain, and Fondo Europeo de Desarrollo Regional [FEDER]). JA is the recipient of an Ajut 2014SGR-4 award (Generalitat de Catalunya). DC was recipient of a predoctoral fellowship from the Spanish Ministry of Education. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Petrezsélyová, S.; López-Malo, M.; Canadell, D.; Roque, A.; Serra-Cardona, A.; Marques Romero, MC.; Vilaprinyó, E.... (2016). Regulation of the Na+/K+-ATPase Ena1 Expression by Calcineurin/Crz1 under High pH Stress: A Quantitative Study. PLoS ONE. 11(6):e0158424-e0158424. https://doi.org/10.1371/journal.pone.0158424Se0158424e015842411

Potassium and Sodium Transport in Yeast

[EN] As the proper maintenance of intracellular potassium and sodium concentrations is vital for cell growth, all living organisms have developed a cohort of strategies to maintain proper monovalent cation homeostasis. In the model yeast Saccharomyces cerevisiae, potassium is accumulated to relatively high concentrations and is required for many aspects of cellular function, whereas high intracellular sodium/potassium ratios are detrimental to cell growth and survival. The fact that S. cerevisiae cells can grow in the presence of a broad range of concentrations of external potassium (10 M–2.5 M) and sodium (up to 1.5 M) indicates the existence of robust mechanisms that have evolved to maintain intracellular concentrations of these cations within appropriate limits. In this review, current knowledge regarding potassium and sodium transporters and their regulation will be summarized. 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Priority of production units based on a climate perspective

On behalf of Umeå Energi, a study was carried out on how the company can reduce its carbon dioxide emissions from district heating production. Umeå Energi has fifteen production units in Umeå with different types of fuel, which supply the city with district heating. The heat demand governs how many of these units that are running at the same time and the order in which these units start is governed by the company priority. The purpose of this project was to investigate how Umeå Energy can reduce its carbon dioxide emissions by changing the order of priority for these production units. The project has developed four scenarios with different priority conditions and the results shows how much the company reduces its emissions by following these different priority schemes. The presently used priority scheme is planned according to economy, but in the scenarios developed in the project, the priority scheme is controlled from a climate perspective, where the production units carbon dioxide emissions control the prioritization. The scenarios gave mixed results. In the best-case scenario Umeå Energi can reduce its carbon dioxide emissions by almost 80%, but then incineration is de-prioritized, which leads to other problems for the society. In the scenarios where incineration is prioritized, the result does not have as great an impact, but still shows that Umeå Energi can reduce its carbon dioxide emissions significantly by changing the production planning.På uppdrag av Umeå Energi gjordes en undersökning om hur företaget kan minska sina koldioxidutsläpp i samband med fjärrvärmeproduktion. Företaget har femton olika produktionsenheter runt om i Umeå med olika typer av bränsle som förser staden med fjärrvärme. Värmebehovet styr hur många av dessa anläggningar som är i gång samtidigt och i vilken ordning dessa enheter startar styrs av företagets prioriteringslista. Syftet med detta examensarbete var att undersöka hur Umeå Energi kan minska sina koldioxidutsläpp genom att ändra prioriteringsordningen för dessa produktionsenheter. Projektet har tagit fram fyra scenarion med olika prioriteringsvillkor och kommit fram till hur mycket företaget minskar sina utsläpp genom att följa dessa olika listor. I dagsläget är denna prioriteringsordning planerad efter ekonomi, men i de scenarier som är framtagna i projektet är prioriteringsordningen mer eller mindre styrda utifrån ett klimatperspektiv, där produktionsenheternas koldioxidutsläpp styr prioriteringen. De olika scenarierna gav blandade resultat, i det bästa scenariot kan företaget minska sina koldioxidutsläpp med ungefär 80 %, men då är avfallsförbränningen bortprioriterad vilket leder till andra problem för samhället. I scenarierna där avfallsförbränningen är prioriterad först ger inte resultatet lika stort genomslag, men visar ändå att Umeå energi kan minska sina koldioxidutsläpp markant genom att ändra på produktionsplaneringen.