Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Comparative transcriptome analysis of Saccharomyces cerevisiae wild type and M-NM-^Tasc1 strain

ABSTRACT: Asc1p and its essential orthologues in higher eukaryotes, as e.g. RACK1 in mammals, are involved in several distinct cellular signaling processes, but the implications of a total deletion have never been assessed in a comprehensive genome-wide manner. This study reveals the major cellular processes affected in a Saccharomyces cerevisiae M-NM-^Tasc1 deletion background via de novo proteome and transcriptome analysis, as well as subsequent phenotypical characterizations. The deletion of ASC1 reduces iron-uptake and causes nitrosative stress, both known indicators for hypoxia, which manifests in a shift of energy metabolism from respiration to fermentation in the M-NM-^Tasc1 strain. The impact of Asc1p in cellular metabolism is further expressed by its regulative role in the MAP kinase signal transduction pathways of invasive/filamentous growth, pheromone response and cell wall integrity. In the M-NM-^Tasc1 mutant strain aberrations from the expected cellular response, mediated by these pathways, can be observed and are linked to changes in protein abundances of pathway-targeted transcription factors. Evidence for the translational regulation of such transcription factors suggests that ribosomal Asc1p is involved in signal transduction pathways by controlling the biosynthesis of the respective final transcriptional regulators. The transcriptional data results from from 3 biological replicates for each strain, whereas for each biological replicate 2 technical replications and dye-swaps were analyzed.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Claudia Pommerenke

PROVIDER: E-GEOD-34286 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

RACK1/Asc1p, a ribosomal node in cellular signaling.

Rachfall Nicole N Schmitt Kerstin K Bandau Susanne S Smolinski Nadine N Ehrenreich Armin A Valerius Oliver O Braus Gerhard H GH

Molecular & cellular proteomics : MCP 20121015 1

RACK1/Asc1p and its essential orthologues in higher eukaryotes, such as RACK1 in metazoa, are involved in several distinct cellular signaling processes. The implications of a total deletion have never been assessed in a comprehensive manner. This study reveals the major cellular processes affected in a Saccharomyces cerevisiae Δasc1 deletion background via de novo proteome and transcriptome analysis, as well as subsequent phenotypical characterizations. The deletion of ASC1 reduces iron uptake a ...[more]

PMID: 23071099

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Project description:The obligatory aerobic acetic acid bacterium Gluconobacter oxydans 621H oxidizes sugars and sugar alcohols primarily in the periplasm and only a small fraction is metabolized in the cytoplasm. The latter can occur either via the Entner-Doudoroff pathway (EDP) or via the pentose phosphate pathway (PPP). The Embden-Meyerhof pathway is non-functional and a cyclic operation of the tricarboxylic acid cycle is prevented by the absence of succinate dehydrogenase. In this work, the cytoplasmic catabolism of fructose formed by oxidation of mannitol was analyzed with a M-NM-^Tgnd mutant lacking the oxidative PPP and a M-NM-^Tedd-eda mutant devoid of the EDP. The growth characteristics of the two mutants under controlled conditions with mannitol as carbon source and enzyme activities showed that the PPP is the main route for cytoplasmic fructose catabolism, whereas the EDP is dispensable and even unfavorable. The M-NM-^Tedd-eda mutant (lacking 6-phosphogluconate dehydratase and 2-keto-3-deoxy-6-phosphogluconate aldolase) formed 24% more cell mass than the reference strain. In contrast, deletion of gnd (6-phosphogluconate dehydrogenase) severely inhibited growth and caused a strong selection pressure for secondary mutations inactivating glucose-6-phosphate dehydrogenase, thus preventing fructose catabolism via the EDP, too. These M-NM-^Tgnd zwf* mutants were almost totally disabled in fructose catabolism, but still produced about 14% of the carbon dioxide of the reference strain, possibly by catabolizing substrates from the yeast extract. Overexpression of gnd in the reference strain improved biomass formation in a similar manner as deletion of edd-eda, further confirming the importance of the PPP for cytoplasmic fructose catabolism. The three transcriptome comparisons of M-NM-^Tupp M-NM-^Tgnd zwf* vs. M-NM-^Tupp and of M-NM-^Tupp M-NM-^Tedd-eda vs. M-NM-^Tupp, and of M-NM-^Tupp M-NM-^Tgnd versus M-NM-^Tupp, were repeated independently three times in biological replicates resulting in 3 x 3 hybridizations as termed by sample 1 to 9.

Dataset Information

Comparative transcriptome analysis of Saccharomyces cerevisiae wild type and M-NM-^Tasc1 strain

Publications

RACK1/Asc1p, a ribosomal node in cellular signaling.

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