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Polyol stress: time course

ABSTRACT: It is unclear how the amount of active nuclear MAPK over time quantitatively affects transcription. Here, we seek to address this issue by studying signal transduction and transcriptional response in a system that separates signalling from adaptation and hence signal strength from signal duration. The system is based on Saccharomyces cerevisiae osmoadaptation and allows modulation of the period of HOG-dependent responses without changing the initial stress intensity. The conditional osmotic stress system includes (i) a yeast mutant (gpd1 gpd2) unable to produce its main osmolyte glycerol, subsequently stressed with (ii) a stress inductor (polyols of different sizes) and allowed to adapt by (iii) expression of polyol flux-mediating aquaglyceroporin (rat AQP9). As there is no endogenous glycerol production, the osmoadaptation rate depends only on the size dependent equilibration rate over the plasma membrane of the polyol used as both stress inductor and compatible solute. Hence, we apply initially identical stresses but with differential durations, and determine the global transcriptional response. Saccharomyces cerevisiae W303-1A (MATa leu2-3/112 ura3-1 trp1-1 his3-11/15 gpd1::TRP1 gpd2::URA3) osmotically stressed with 1M of glycerol, erythritol, xylitol or sorbitol. Samples are taken in triplicates (except for sorbitol 20 an 90 min and 20 min xylitol were duplicates were taken) in time course series after stress application. Total of 45 samples. RNA from from cultures of gpd1 gpd2 cells expressing rAQP9 prior to polyol exposure was used as reference RNA for all microarrays.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Abraham Ericsson

PROVIDER: E-GEOD-29716 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Polyol stress: time course

Project description:It is unclear how the amount of active nuclear MAPK over time quantitatively affects transcription. Here, we seek to address this issue by studying signal transduction and transcriptional response in a system that separates signalling from adaptation and hence signal strength from signal duration. The system is based on Saccharomyces cerevisiae osmoadaptation and allows modulation of the period of HOG-dependent responses without changing the initial stress intensity. The conditional osmotic stress system includes (i) a yeast mutant (gpd1 gpd2) unable to produce its main osmolyte glycerol, subsequently stressed with (ii) a stress inductor (polyols of different sizes) and allowed to adapt by (iii) expression of polyol flux-mediating aquaglyceroporin (rat AQP9). As there is no endogenous glycerol production, the osmoadaptation rate depends only on the size dependent equilibration rate over the plasma membrane of the polyol used as both stress inductor and compatible solute. Hence, we apply initially identical stresses but with differential durations, and determine the global transcriptional response.

2014-05-31 | GSE29716 | GEO

Genome-Wide expression under osmotic stress conditions in Saccharomyces cerevisiae

Project description:Changes in RNA levels during osmotic stress were investigated. Total RNA was extracted from a wild-type yeast strain before and after treatment with 0.4 M NaCl and the corresponding cDNAs were hybridazed on Tiling arrays. In particular, for all the intron-containing genes, the changes in the levels of intron signal in stressed cells related to the intron signal in the non-stressed cells, and the changes in the levels of exon signal in stresses cells related to the exon signal in non-stressed cells were investigated. The supplementary bar file contains the ratios between stress signals respect to non-stress signals, using the average of the 3 biological replicas.

2013-01-03 | GSE43236 | GEO

Histone H3 T11 phosphorylation by Sch9 and CK2 regulates lifespan by controlling the nutritional stress response [ChIP-seq]

Project description:We report the changes of Histone H3 T11 phosphorylation (H3pT11) signal upon nutritional stress (YP with 3% glycerol).

2018-06-26 | GSE111218 | GEO

Transcription profiling of yeast strains lacking Met4 or Met4 cofactors at time points after sulfur limitation

Project description:All yeast strains used in this study (Table 1) are in the W303 background (ade2-1 can1-100, his3-1,15 leu2-3,112 trp1-1 ura3). For sulfur limitation microarray studies, WT, met4 delete, met31 delete met32 delete, cbf1 delete, and met28 delete strains were grown in minimal B-media [see Cherest, H., and Surdin-Kerjan, Y. (1992). Genetic analysis of a new mutation conferring cysteine auxotrophy in Saccharomyces cerevisiae: updating of the sulfur metabolism pathway. Genetics 130, p51-58 for B-media composition] supplemented with 0.5mM methionine as the sole sulfur source. An aliquot of cells was harvested for a t=0 time point while the remainder were filtered through a .22um Stericup filter (Millipore), then washed and resuspended in pre-warmed (30 C) B-media lacking any source of sulfur. Cells were harvested after 20, 40, and 80 minutes.

2009-11-30 | E-MEXP-2429 | biostudies-arrayexpress

3′-seq of Saccharomyces cerevisiae grown in eight different conditions

Project description:Saccharomyces cerevisiae strain BY4741 (MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0) was grown in YPD medium. For control conditions, cells were grown in mid-log phase at 30°C for 20 min or 2 hours. Stress conditions were heat shock (37°C for 20 min), diauxic shift (harvested at 2 hours after glucose depletion), DNA damage (harvested after 2 hours of treatment with 5 µg/ml 4-nitroquinoline 1-oxide (Sigma)). Moreover, four conditions of the yeast metabolic cycle were analyzed. S. cerevisiae strain CEN.PK (MATa URA3 TRP1 LEU2 HIS3 MAL2-8C SUC2) was used and cultivated as described elsewhere (Nocetti and Whitehouse, Genes Dev, 2016). The period of metabolic oscillation was approximately 3.2 h. Cells were harvested at 16, 32, 48, and 60 min from the start of each cycle for one oxidative, two reductive/building, and one reductive/charging phase points. Total RNA from each sample was extracted by the hot phenol method and used for library preparation. 3′-seq libraries were prepared as described previously (Lianoglou et al., Genes Dev, 2013). Each condition was sequenced in three biological replicates. The 27 individual libraries were pooled and subjected to sequencing with HiSeq with SR50 at the Integrated Genomics Operation (MSKCC).

2023-09-30 | GSE213720 | GEO

Hg stress

Project description:Effect of HgCl2-treatment (dose-response:15min, 30min, 2h, and 24h) on cellular metabolism, resp on transcription. Differences between BY-4741 (MATa leu2 ura3 his3 met15 can1) + BYstb5 (MATa stb5::canMX4) to gain new insights into Hg-toxicity itself and adaptation to Hg-stress in yeast Saccharomyces cerevisiae.

2011-05-01 | E-MEXP-2991 | biostudies-arrayexpress

Transcription profiling of yeast mutant strains vs WT: YIA29 (mdl1::HIS3); YIA30 (yme1::KAN); YIA31 (mdl1::HIS3 yme1::KAN);

Project description:All yeast strains were in the genetic background W303-1A (MATa leu2-3,112 trp1-1 ura3-1 can1-100 ade2-1 his3-11,15). Gene expression profiles of the following mutant strains were compared with WT: YIA29 (mdl1::HIS3); YIA30 (yme1::KAN); YIA31 (mdl1::HIS3 yme1::KAN); 2 independent RNA preparations per strain were used (A and B); 2 independent array hybridisations per RNA preparation were performed (color switch experiments).

2005-12-31 | E-EMBL-5 | biostudies-arrayexpress

LC-MS confirmation of alternative YPD1 isoforms from Magnaporthe oryzae

Project description:This project aims to understand better the stress response mechanisms of components of the well known High Osmolarity Glycerol two-component regulatory system in the model organism Magnaporthe oryzae. Currently, two putative isoforms of the transfer protein YPD1 (MGG_07173) are described as theoretical proteins in the UniProt database (G4MTL0 and G4MTK9). For better understanding of signal transduction and response triggering we aimed to identify alternative isoforms that might explain the variety of triggered responses throughout the YPD1 signal transfer hub.

2021-09-10 | PXD024832 | Pride

E. coli BW25113 frdC vs. E. coli BW25113 HW2

Project description:Glycerol is an attractive feedstock for biofuels since it accumulates as a byproduct during biodiesel operations; hence, it is interesting to consider converting glycerol to hydrogen using the formate hydrogen lyase system of Escherichia coli which converts pyruvate to hydrogen. Starting with Escherichia coli BW25113 frdC that lacks fumarate reductase to eliminate the negative effect of accumulated hydrogen on glycerol fermentation and by using both adaptive evolution and chemical mutagenesis combined with a selection method based on increased growth on glycerol, we obtained an improved strain, HW2, that produces 20-fold more hydrogen in glycerol medium (0.68 mmol/L/h) compared to that of frdC mutant. HW2 also grows 5-fold faster (0.25 1/h) than BW25113 frdC on glycerol, so it achieves a reasonable growth rate. Corroborating the increase in hydrogen production, glycerol dehydrogenase activity in HW2 increased 4-fold compared to BW25113 frdC. In addition, a whole-transcriptome study revealed that several pathways that would decrease hydrogen yields were repressed in HW2 (fbp, focA, and gatYZ) while a beneficial pathway, eno which encodes enolase was induced.

2010-10-31 | GSE18972 | GEO

Response of D. hansenii to osmotic stress

Project description:The euryhaline marine yeast Debaromyces hansenii is a model system for the study of genes related to osmotic stress. To study the transcriptional response of this organism to osmotic stress we have used the two color microarray based gene expression analysis of 6211 genes which constitutes the whole genome. Analysis was done at three time points after induction with salt (0.5h, 3h and 6h.) The mRNA level of 64 genes significantly increased at least 3-fold after induction with 2M NaCl whereas that of 45 genes was 3-fold diminished. The induced as well as the repressed genes were grouped into functional categories to identify biochemical processes possibly affected by osmotic shock. 53% of the induced genes encode for ribosomal proteins, 12.5% for mitochondrial and redox, 6% for aminoacid, cell wall and carbohydrate, and 1.5%for heat shock, protein transport and glycerol proteins. The function of 31% of repressed genes is currently unknown. 15% of the repressed genes are involved in carbohydrate metabolism and protective function, 6% are associated with cell wall, redox and signal transduction, and 4% in vacuolar and lipid functions. Surprisingly, the activity of NAD+ glycerol 3-phosphate dehydrogenase gene (GPD1) involved in the glycerol biosynthesis in response to osmotic stress did not show induction. Keywords: time course, stress response, mRNA expression

2008-03-15 | GSE9911 | GEO

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