Project description:The expression of plant defensins in Saccharomyces cerevisiae has been shown to increase tolerance to Zn of yeast (Mirouze et al., 2006; Shahzad et al., 2013). The present transcriptomic approach has been conducted to uncover the mechanism underlying this phenotype. WT yeast was compared to yeast expressing the AhPFF1.1b plant defensin when grown in both control medium and medium supplemented with 20 mM ZnSO4. 4 conditions with 2 biological replicates compared: control yeast (BY4741) grown on control medium, control yeast grown in the presence of 20 mM ZnSO4, yeast expressing the AhPDF1.1b defensin grown in control medium, yeast expressing the AhPDF1.1b defensin grown in the presence of 20 mM ZnSO4
Project description:The expression of plant defensins in Saccharomyces cerevisiae has been shown to increase tolerance to Zn of yeast (Mirouze et al., 2006; Shahzad et al., 2013). The present transcriptomic approach has been conducted to uncover the mechanism underlying this phenotype. WT yeast was compared to yeast expressing the AhPFF1.1b plant defensin when grown in both control medium and medium supplemented with 20 mM ZnSO4.
Project description:Zinc is indispensable for the catalytic activity and structural stability of many proteins, and its deficiency can have severe consequences for microbial growth in natural and industrial environments. For example, Zn depletion in wort negatively affects beer fermentation and quality. Several studies have investigated yeast adaptation to low Zn supply, but were all performed in batch cultures, where specific growth rate depends on Zn availability. The transcriptional responses to growth-rate and Zn availability are then intertwined, which obscures result interpretation. In the present study, transcriptional responses of Saccharomyces cerevisiae to Zn availability were investigated at a fixed specific growth rate under Zn limitation and excess in chemostat culture. To investigate the context-dependency of this transcriptional response, yeast was grown under several chemostat regimes resulting in various carbon (glucose), nitrogen (ammonium) and oxygen supplies. A robust set of genes that responded consistently to Zn limitation was identified and enabled the definition of a Zn-specific Zap1 regulon comprising of 26 genes and characterized by a broader ZRE consensus (MHHAACCBYNMRGGT) than so far described. Most surprising was the Zn-dependent regulation of genes involved in storage carbohydrate metabolism. Their concerted down-regulation was physiologically relevant as revealed by a substantial decrease in glycogen and trehalose cellular content under Zn limitation. An unexpectedly large amount of genes were synergistically or antagonistically regulated by oxygen and Zn availability. This combinatorial regulation suggested a more prominent involvement of Zn in mitochondrial biogenesis and function than hitherto identified Keywords: Chemostat based transcriptome analysis
Project description:Zinc is indispensable for the catalytic activity and structural stability of many proteins, and its deficiency can have severe consequences for microbial growth in natural and industrial environments. For example, Zn depletion in wort negatively affects beer fermentation and quality. Several studies have investigated yeast adaptation to low Zn supply, but were all performed in batch cultures, where specific growth rate depends on Zn availability. The transcriptional responses to growth-rate and Zn availability are then intertwined, which obscures result interpretation. In the present study, transcriptional responses of Saccharomyces cerevisiae to Zn availability were investigated at a fixed specific growth rate under Zn limitation and excess in chemostat culture. To investigate the context-dependency of this transcriptional response, yeast was grown under several chemostat regimes resulting in various carbon (glucose), nitrogen (ammonium) and oxygen supplies. A robust set of genes that responded consistently to Zn limitation was identified and enabled the definition of a Zn-specific Zap1 regulon comprising of 26 genes and characterized by a broader ZRE consensus (MHHAACCBYNMRGGT) than so far described. Most surprising was the Zn-dependent regulation of genes involved in storage carbohydrate metabolism. Their concerted down-regulation was physiologically relevant as revealed by a substantial decrease in glycogen and trehalose cellular content under Zn limitation. An unexpectedly large amount of genes were synergistically or antagonistically regulated by oxygen and Zn availability. This combinatorial regulation suggested a more prominent involvement of Zn in mitochondrial biogenesis and function than hitherto identified Keywords: Chemostat-based transcriptome analysis
Project description:Zinc is indispensable for the catalytic activity and structural stability of many proteins, and its deficiency can have severe consequences for microbial growth in natural and industrial environments. For example, Zn depletion in wort negatively affects beer fermentation and quality. Several studies have investigated yeast adaptation to low Zn supply, but were all performed in batch cultures, where specific growth rate depends on Zn availability. The transcriptional responses to growth-rate and Zn availability are then intertwined, which obscures result interpretation. In the present study, transcriptional responses of Saccharomyces cerevisiae to Zn availability were investigated at a fixed specific growth rate under Zn limitation and excess in chemostat culture. To investigate the context-dependency of this transcriptional response, yeast was grown under several chemostat regimes resulting in various carbon (glucose), nitrogen (ammonium) and oxygen supplies. A robust set of genes that responded consistently to Zn limitation was identified and enabled the definition of a Zn-specific Zap1 regulon comprising of 26 genes and characterized by a broader ZRE consensus (MHHAACCBYNMRGGT) than so far described. Most surprising was the Zn-dependent regulation of genes involved in storage carbohydrate metabolism. Their concerted down-regulation was physiologically relevant as revealed by a substantial decrease in glycogen and trehalose cellular content under Zn limitation. An unexpectedly large amount of genes were synergistically or antagonistically regulated by oxygen and Zn availability. This combinatorial regulation suggested a more prominent involvement of Zn in mitochondrial biogenesis and function than hitherto identified. Keywords: Chemostat-based transcriptome analysis
Project description:Paired-end sequencing study of nucleosomal DNA prepared from budding yeast by micrococcal nuclease digestion. Comparison of control cells with cells treated with 10 mM 3-aminotriazole for 20 minutes
Project description:In this study we investigated the transcriptional response of the yeast Saccharomyces cerevisiae to potassium starvation. To this end yeast cells were grown for 60 min in media without potassium or in media with a standard potassium concnetration (50 mM KCl). Using Serial Analysis of Gene Expression (SAGE)-tag sequencing the effect of potassium starvation on the transcriptome was determined.
Project description:The model yeast species Saccharomyces cerevisiae is used in many fundamental and applied research applications, including biosensors and production of many compounds. However, given the enormous work invested in the studies of yeast transcription response to various conditions, there are still substances not explored in this regard. In this work, we explore the transcriptional response of S. cerevisiae to a wide range of concentrations of the D-enantiomer of lactic acid and compare it to the response to L-lactic acid. Of these conditions, we only recorded a transcriptional response to the relatively high concentrations of DLA of 5 and 45 mM, as well as to 45 mM LLA. Our data did not reveal any natural yeast promoters that quantitatively sense D-lactic acid but provide the first description of the transcriptome-wide response to DLA, as well as enrich our understanding of the LLA response.