Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcriptional profile of an eca39∆ mutant fission yeast

ABSTRACT: An branched-chain amino acids auxotroph eca39∆ mutant fission yeast exhibits an unusual adaptive growth phenotype on solid minimal media containing Ile, Leu and Val when other strains are growing nearby. The transcriptional profiles of an eca39∆ mutant before and after the adaptation were determined using Affymetrix DNA microarrays. Wild-type, the eca39∆ mutant, and the adapted eca39∆ mutant fission yeasts were inoculated in YE+2mM Ile, Leu and Val, and harvested at OD ~ 1. Total RNAs were purified from the 3 samples.

ORGANISM(S): Schizosaccharomyces pombe

SUBMITTER: Hidekazu Takahashi

PROVIDER: E-GEOD-29355 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

ACCESS DATA

Publications

The SAGA histone acetyltransferase complex regulates leucine uptake through the Agp3 permease in fission yeast.

Takahashi Hidekazu H Sun Xiaoying X Hamamoto Makiko M Yashiroda Yoko Y Yoshida Minoru M

The Journal of biological chemistry 20120918 45

Metabolic responses of unicellular organisms are mostly acute, transient, and cell-autonomous. Regulation of nutrient uptake in yeast is one such rapid response. High quality nitrogen sources such as NH(4)(+) inhibit uptake of poor nitrogen sources, such as amino acids. Both transcriptional and posttranscriptional mechanisms operate in nutrient uptake regulation; however, many components of this system remain uncharacterized in the fission yeast, Schizosaccharomyces pombe. Here, we demonstrate t ...[more]

PMID: 22992726

Similar Datasets

Project description:Transcription profiling of wild type, relA-, and relA-spoT-, crp-, dksA-, rpoS-, lrp- mutant strains of E. coli starved for isoleucine Bacteria comprehensively reorganize their global gene expression when faced with nutrient exhaustion. In Escherichia coli and other free-living bacteria, the alarmone ppGpp facilitates this massive response by directly or indirectly coordinating the down-regulation of genes of the translation apparatus, and the induction of biosynthetic genes and the general stress response. Such a large reorientation likely requires the cooperative activities of many different genetic regulators, yet the structure of the transcription network below the level of ppGpp remains poorly defined. Using isoleucine starvation as an experimental model system for amino acid starvation, we identified genes that required ppGpp, Lrp, and RpoS for their induction. Surprisingly, despite the fact that the overwhelming majority of genes controlled by Lrp and RpoS required ppGpp for their activation, we found that these two regulons were not induced simultaneously. The data reported here suggest that metabolic genes, such as those of the Lrp regulon, require only a low basal level of ppGpp for their efficient induction. In contrast, the RpoS-dependent general stress response is not robustly induced until relatively high levels of ppGpp accumulate. Here we describe a data-driven conceptual model that explains how bacterial cells allocate transcriptional resources between metabolic and stress survival processes by discretely tuning regulatory activities to a central indicator of cellular physiology. The regulatory structure that emerges is consistent with a rheostatic model of the stringent response that allows cells to efficiently adapt to a wide range of nutritional environments. Keywords: genetic modification design; stress response; isoleucine starvation Two experiments were run. First experiment: WT and several mutant strains were starved for isoleucine (exhaustion of 60 uM ile). 40 minutes after starvation, RNA was extracted. All samples were compared to RNA from rapidly growing WT cells in identical medium replete with isoleucine (400 uM). 16 samples were hybridized: duplicates of 8 strains/conditions. Wildtype in log phase (OD = 0.4) with replete ile was control for ile starved samples. Second experiment: WT strain was starved for isoleucine (exhaustion of 60 uM ile). RNA was extracted at 12 timepoints as the cells entered stationary phase. All samples were compared to RNA from rapidly growing WT cells in identical medium replete with isoleucine (400 uM). 14 samples were hybridized: duplicates of the control condition and single timepoints during starvation. Wildtype in log phase (OD = 0.4) with replete ile was control for ile starved samples.

Project description:Trypanosoma cruzi, the etiological agent of Chagas disease, has a complex life cycle occurring between triatomine insects (responsible for the vectorial transmission of the infection) and mammals. Inside the insect vector, epimastigote forms (proliferative, non-infective to mammals) differentiate into metacyclic trypomastigotes forms (non-proliferative, infective to mammals). This differentiation process happens at the final portion of insect vector’s gut and it is crucial for the progress of T. cruzi life cycle. Factors that play a role in metacyclogenesis include temperature, pH and nutritional stress followed by availability of specific metabolites. Previous results showed that some amino acids such as Pro, His and Gln are able to fully support cell differentiation whereas the BCAAs completely block it. In addition, combination of the “pro-metacyclogenic” amino acid Pro with one of the “anti-metacyclogenic” BCAAs produces viable metacyclics, however, with a drastically reduced infectivity. In order to investigate the peculiarities of metacyclic parasites differentiated in the presence of BCAAs, we performed proteomics analyses to compare metacyclics obtained in triatomine artificial urine (TAU) supplemented with Pro alone and in combination with Leu, Ile or Val. Our analyses revealed that when compared to the metacyclics originated in TAU-Pro, 40 proteins are differentially regulated in metacyclics from TAU-Pro-Leu, whereas 131 and 179 are differentially regulated in TAU-Pro-Ile and TAU-Pro-Val, respectively. From each group, 22, 11 and 13% of the proteins are putatively related to metabolic processes, respectively. Overall, enzymes involved in both glycolysis and TCA cycle are reduced in metacyclics obtained in the presence of Pro-BCAAs whereas enzymes involved in amino acid and purine metabolic pathways are increased. In addition, metacyclics obtained in the presence of Pro-Ile and Pro-Val have enzymes involved in lipid and redox metabolism increased. Together our analyses indicate that metacyclics obtained in the presence of BCAA can have their metabolism reshaped, which in turn can contribute to the reduced infectivity observed in these parasites.

Dataset Information

Transcriptional profile of an eca39∆ mutant fission yeast

Publications

The SAGA histone acetyltransferase complex regulates leucine uptake through the Agp3 permease in fission yeast.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets