Saccharomyces cerevisiae
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA292121 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR2148150.fastq.gz | Fastqsanger.gz | |||
| SRR2148160.fastq.gz | Fastqsanger.gz | |||
| SRR2148170.fastq.gz | Fastqsanger.gz | |||
| SRR2148180.fastq.gz | Fastqsanger.gz | |||
| SRR2148190.fastq.gz | Fastqsanger.gz |
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Project description:The yeast mating pathway serves as a valuable model of eukaryotic mitogen activated protein kinase (MAPK) pathways, several components of which are actual and potential therapeutic drug targets. Here we present a new screening method we developed that combines two technologies, fluorescence activated cell sorting (FACS) with Barcode analysis by Sequencing (Bar-Seq). Using the new screening method and pFUS1-GFP as a reporter, we identified known mutants in the mating pathway and also characterized a source of false positives that arose as a result of the design of this screen. Additionally from the screen, we identified that a deletion mutant, sub1Δ, that exhibits increased expression of the GFP reporter. Our ChIP-Seq data shows that Sub1 increases binding to the promoters of several pheromone-inducible genes and supports our finding that SUB1 is involved in regulating the expression of genes in mating pathway. RNA-Seq of a sub1Δ mutant shows that the majority of genes have no change in RNA and of the small subset that do, most of these show a decrease in expression. In this study, we find that the sub1Δ mutant has increased basal levels of a small subset of genes including IMD2 (consistent with previous reports) and a gene necessary for efficient mating, FIG1.
2016-05-09 | GSE71813 | GEO
Project description:Haploid budding yeast has two mating types, defined by the alleles of the MAT locus, MATa and MATα. Mating occurs when two haploid cells of opposite mating types signal to each other using reciprocal pheromones and receptors, polarize and grow towards each other, and eventually fuse to form a single diploid cell. The pheromones and receptors are necessary and sufficient to define a mating type, but other mating type-specific proteins make mating more efficient. We examined the role of these proteins by genetically engineering “transvestite” cells that swap the pheromone, pheromone receptor, and pheromone processing factors of one mating type for another. These cells can mate with each other, but their mating is inefficient. By characterizing their mating defects and examining their transcriptomes, we found Afb1 (a-factor barrier), a novel MATα-specific protein that interferes with a-factor, the pheromone secreted by MATa cells. We show that strong pheromone secretion is essential for efficient mating and that the weak mating of transvestites can be improved by boosting their pheromone production. Using synthetic biology, it is possible to characterize the factors that control efficiency in biological processes. In the case of budding yeast mating, selection for increased mating efficiency is likely to have continually boosted pheromone levels and the ability to discriminate between partners who make more (potentially fitter) and less (potentially less fit) pheromones. This sensitivity to which partner makes more pheromone comes at a cost: it means mating is not robust in situations where all potential partners make less pheromone.
2014-01-07 | GSE49372 | GEO
Project description:Yeast deletion pool Barcode-sequencing
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Project description:The target of rapamycin (TOR) plays a central role in eukaryotic cell growth control. With prevalent hyper-activation of the mTOR pathway in human cancers, novel strategies to enhance TOR pathway inhibition are highly desirable. We used a yeast-based high-throughput chemical genetic screen to identify small-molecule enhancers of rapamycin (SMERs) and used whole genome expression analysis to identify their mechanisms of action.
2011-06-30 | GSE22270 | GEO
Project description:High throughput yeast replicative lifespan screen uncovers histone deacetylase complex HDA as novel regulator of aging
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Project description:The target of rapamycin (TOR) plays a central role in eukaryotic cell growth control. With prevalent hyper-activation of the mTOR pathway in human cancers, novel strategies to enhance TOR pathway inhibition are highly desirable. We used a yeast-based high-throughput chemical genetic screen to identify small-molecule enhancers of rapamycin (SMERs) and used whole genome expression analysis to identify their mechanisms of action. We incubated Met30 temperature-sensitive yeast strain at either the permissive (room temperature) or non-permissive (35°C) temperature for 1 hour prior to RNA extraction and hybridization on Affymetrix microarrays. Comparison of expression profiles enabled identification of gene-signature characteristic of Met30 inhibition.
2011-06-30 | E-GEOD-22270 | biostudies-arrayexpress
Project description:The target of rapamycin (TOR) plays a central role in eukaryotic cell growth control. With prevalent hyper-activation of the mTOR pathway in human cancers, novel strategies to enhance TOR pathway inhibition are highly desirable. We used a yeast-based high-throughput chemical genetic screen to identify small-molecule enhancers of rapamycin (SMERs) and used whole genome expression analysis to identify their mechanisms of action.
2010-06-28 | GSE22269 | GEO
Project description:We examined the mating response of three yeast species, by subjecting a-type cells from each of them to alpha-factor (6uM). The expression of cells before and after (60min) addition of alpha-factor were compared using S. cerevisiae cDNA microarrays. In addition, we verified our results with a true mating experiment: the expression of a-type and alpha-type cells from S. cerevisiae were compared before and after mixing them for 90 minutes (i.e. allowing them to mate). Keywords: yeast mating response, comparison among species
2007-05-20 | GSE7525 | GEO
Project description:The target of rapamycin (TOR) plays a central role in eukaryotic cell growth control. With prevalent hyper-activation of the mTOR pathway in human cancers, novel strategies to enhance TOR pathway inhibition are highly desirable. We used a yeast-based high-throughput chemical genetic screen to identify small-molecule enhancers of rapamycin (SMERs) and used whole genome expression analysis to identify their mechanisms of action. We treated yeast individually with SMERs 1-5, rapamycin, or DMSO (solvent control) for 30 minutes prior to RNA extraction and hybridization on Affymetrix microarrays. Expression profiles of SMER-treated samples were compared to that of DMSO (solvent control) and rapamycin-treated samples to identify gene expression signatures unique to SMER-treated samples.
2010-07-29 | E-GEOD-22269 | biostudies-arrayexpress
Project description:Haploid budding yeast has two mating types, defined by the alleles of the MAT locus, MATa and MATM-NM-1. Mating occurs when two haploid cells of opposite mating types signal to each other using reciprocal pheromones and receptors, polarize and grow towards each other, and eventually fuse to form a single diploid cell. The pheromones and receptors are necessary and sufficient to define a mating type, but other mating type-specific proteins make mating more efficient. We examined the role of these proteins by genetically engineering M-bM-^@M-^\transvestiteM-bM-^@M-^] cells that swap the pheromone, pheromone receptor, and pheromone processing factors of one mating type for another. These cells can mate with each other, but their mating is inefficient. By characterizing their mating defects and examining their transcriptomes, we found Afb1 (a-factor barrier), a novel MATM-NM-1-specific protein that interferes with a-factor, the pheromone secreted by MATa cells. We show that strong pheromone secretion is essential for efficient mating and that the weak mating of transvestites can be improved by boosting their pheromone production. Using synthetic biology, it is possible to characterize the factors that control efficiency in biological processes. In the case of budding yeast mating, selection for increased mating efficiency is likely to have continually boosted pheromone levels and the ability to discriminate between partners who make more (potentially fitter) and less (potentially less fit) pheromones. This sensitivity to which partner makes more pheromone comes at a cost: it means mating is not robust in situations where all potential partners make less pheromone. 4 conditions were analysed, each with 3 biological replicates. The conditions were unstimulated MATa cells in YPD. Stimulated MATa cells in YPD+10nM M-NM-1-factor. Unstimulated MATM-NM-1 cells in YPD. Stimulated MATM-NM-1 cells in YPD+10nM M-NM-1-factor.
2014-01-07 | E-GEOD-49372 | biostudies-arrayexpress