Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptional response of Saccharomyces cerevisiae to potassium starvation

ABSTRACT: 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. 4 samples of cells grown in media without potassium and 4 samples of cells grown in the presence of potassium were analyzed.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Paul van Heusden

PROVIDER: E-GEOD-57093 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Transcriptional response of Saccharomyces cerevisiae to potassium starvation.

Anemaet Ida G IG van Heusden G Paul H GP

BMC genomics 20141129

<h4>Background</h4>Ion homeostasis is essential for every cell and aberrant cation homeostasis is related to diseases like Alzheimer's disease and epilepsy. The mechanisms responsible for cation homeostasis are only partly understood. The yeast Saccharomyces cerevisiae is an excellent organism to study fundamental aspects of cation homeostasis. In this study we investigated the transcriptional response of this yeast to potassium starvation by using Serial Analysis of Gene Expression (SAGE)-tag s ...[more]

PMID: 25432801

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Project description:Background. Most methods for constructing aneuploid yeast strains that have gained a specific chromosome rely on spontaneous failures of cell division fidelity. In Saccharomyces cerevisiae, extra chromosomes can be obtained when errors in meiosis or mitosis lead to nondisjunction, or when nuclear breakdown occurs in heterokaryons. We describe a strategy for constructing N+1 disomes that does not require such spontaneous failures. The method combines two well-characterized genetic tools: a conditional centromere that transiently blocks disjunction of one specific chromosome, and a duplication marker assay that identifies disomes among daughter cells. To test the strategy, we targeted chromosomes III, IV, and VI for duplication. Results. The centromere of each target chromosome was replaced by a conditional centromere that can be blocked by growth in galactose, and ura3::HIS3, a duplication marker. Transient exposure to galactose induced the appearance of colonies carrying duplicated markers for chromosomes III or IV, but not VI. Microarray-based comparative genomic hybridization (CGH) confirmed that disomic strains carrying extra chromosome III or IV were generated. Chromosome VI contains several genes known to be deleterious when overexpressed, including the beta-tubulin gene TUB2. To test whether a tubulin stoichiometry imbalance prevents viability in cells carrying an extra chromosome VI, we supplied the parent strain with extra copies of the alpha-tubulin gene TUB1, then induced nondisjunction. Galactose-dependent chromosome VI disomes were produced, as revealed by CGH. Some chromosome VI disomes also carried extra, unselected copies of additional chromosomes. Conclusions. This method causes efficient nondisjunction of a targeted chromosome and allows resulting disomic cells to be identified and maintained. We used the method to test the role of tubulin imbalance in the apparent lethality of disomic chromosome VI. Our results indicate that a tubulin imbalance is necessary for disomic VI lethality, but it may not be the only dosage-dependent effect. Keywords: comparative genomic hybridization, CGH Candidate disomes (Ura+His+) were compared to their euploid parents by CGH. 27 candidate strains were examined with 27 arrays. 4 candidates targeted chromosome III (3 induced, 1 spontaneous), 14 candidates targeted chromosome IV (8 induced, 6 spontaneous), and 9 candidates targeted chromosome VI (all induced).

Dataset Information

Transcriptional response of Saccharomyces cerevisiae to potassium starvation

Publications

Transcriptional response of Saccharomyces cerevisiae to potassium starvation.

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