S288c Quiescent and Non-quiescent replicates
Ontology highlight
ABSTRACT:
ORGANISM(S): Saccharomyces cerevisiae
SUBMITTER: Anthony Aragon
PROVIDER: E-GEOD-8559 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Publications
Characterization of differentiated quiescent and nonquiescent cells in yeast stationary-phase cultures.
Aragon Anthony D AD Rodriguez Angelina L AL Meirelles Osorio O Roy Sushmita S Davidson George S GS Tapia Phillip H PH Allen Chris C Joe Ray R Benn Don D Werner-Washburne Margaret M
Molecular biology of the cell 20080116 3
Cells in glucose-limited Saccharomyces cerevisiae cultures differentiate into quiescent (Q) and nonquiescent (NQ) fractions before entering stationary phase. To understand this differentiation, Q and NQ cells from 101 deletion-mutant strains were tested for viability and reproductive capacity. Eleven mutants that affected one or both phenotypes in Q or NQ fractions were identified. NQ fractions exhibit a high level of petite colonies, and nine mutants affecting this phenotype were identified. Mi ...[more]
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Project description:Cells in glucose-limited Saccharomyces cerevisiae cultures differentiate into quiescent (Q) and non-quiescent (NQ) fractions prior to entering stationary phase. To identify genes involved in this differentiation, Q and NQ cells from 101 deletion-mutant strains were tested for viability and reproductive capacity. Twenty-one mutants were identified, including 7 that affected reproductive capacity of both cell types. Thirteen affected only Q or NQ cells, indicating significant differentiation of these cell types. doa4 strains, lacking ubiquitin hydrolase, affected viability and reproductive capacity in both cell types. More than 1300 mRNAs differentiating Q and NQ cell fractions were identified by microarray analysis. Gene-ontology analysis of Q-cell mRNAs showed significant increases in protein-encoding mRNAs involved in membrane maintenance, oxidative stress response, and signal transduction. NQ-cell mRNAs encode proteins involved in Ty-element transposition and DNA recombination, consistent with apoptosis in these cells. Consistent with preparation for rapid response to environmental stimuli, approximately 2000 protease-labile mRNAs were identified in Q cells. The differentiation of these cell types and the ability of genes to selectively affect the survival of Q or NQ cells in yeast are relevant to chronological aging, cell-cycle, genome-evolution, and stem-cell research and provides insight into complex responses that even simple organisms have to starvation. Samples were separeted into Q and NQ fractions using percoll density gradients. During RNA isolation samples were treated with proteinase K or 10 mM tris. This was done to test of any differences in portease labile mRNAs in Q and NQ cells.
2010-05-27 | E-GEOD-8558 | biostudies-arrayexpress
Project description:Cells in glucose-limited Saccharomyces cerevisiae cultures differentiate into quiescent (Q) and non-quiescent (NQ) fractions prior to entering stationary phase. To identify genes involved in this differentiation, Q and NQ cells from 101 deletion-mutant strains were tested for viability and reproductive capacity. Twenty-one mutants were identified, including 7 that affected reproductive capacity of both cell types. Thirteen affected only Q or NQ cells, indicating significant differentiation of these cell types. doa4 strains, lacking ubiquitin hydrolase, affected viability and reproductive capacity in both cell types. More than 1300 mRNAs differentiating Q and NQ cell fractions were identified by microarray analysis. Gene-ontology analysis of Q-cell mRNAs showed significant increases in protein-encoding mRNAs involved in membrane maintenance, oxidative stress response, and signal transduction. NQ-cell mRNAs encode proteins involved in Ty-element transposition and DNA recombination, consistent with apoptosis in these cells. Consistent with preparation for rapid response to environmental stimuli, approximately 2000 protease-labile mRNAs were identified in Q cells. The differentiation of these cell types and the ability of genes to selectively affect the survival of Q or NQ cells in yeast are relevant to chronological aging, cell-cycle, genome-evolution, and stem-cell research and provides insight into complex responses that even simple organisms have to starvation. Yeast deletion mutants (BY4742 background) were grown to stationary-phase (7 days) and then separted into Q and NQ cells using Percoll density gradients. Microarrays were carried out on these different cell fractions.
2007-12-26 | E-GEOD-8561 | biostudies-arrayexpress
Project description:Cells in glucose-limited Saccharomyces cerevisiae cultures differentiate into quiescent (Q) and non-quiescent (NQ) fractions prior to entering stationary phase. To identify genes involved in this differentiation, Q and NQ cells from 101 deletion-mutant strains were tested for viability and reproductive capacity. Twenty-one mutants were identified, including 7 that affected reproductive capacity of both cell types. Thirteen affected only Q or NQ cells, indicating significant differentiation of these cell types. doa4 strains, lacking ubiquitin hydrolase, affected viability and reproductive capacity in both cell types. More than 1300 mRNAs differentiating Q and NQ cell fractions were identified by microarray analysis. Gene-ontology analysis of Q-cell mRNAs showed significant increases in protein-encoding mRNAs involved in membrane maintenance, oxidative stress response, and signal transduction. NQ-cell mRNAs encode proteins involved in Ty-element transposition and DNA recombination, consistent with apoptosis in these cells. Consistent with preparation for rapid response to environmental stimuli, approximately 2000 protease-labile mRNAs were identified in Q cells. The differentiation of these cell types and the ability of genes to selectively affect the survival of Q or NQ cells in yeast are relevant to chronological aging, cell-cycle, genome-evolution, and stem-cell research and provides insight into complex responses that even simple organisms have to starvation. Yeast deletion mutants (BY4742 background) were grown to stationary-phase (7 days) and then separted into Q and NQ cells using Percoll density gradients. Microarrays were carried out on these different cell fractions.
2007-12-26 | E-GEOD-8560 | biostudies-arrayexpress
Project description:Cells in glucose-limited Saccharomyces cerevisiae cultures differentiate into quiescent (Q) and non-quiescent (NQ) fractions prior to entering stationary phase. To identify genes involved in this differentiation, Q and NQ cells from 101 deletion-mutant strains were tested for viability and reproductive capacity. Twenty-one mutants were identified, including 7 that affected reproductive capacity of both cell types. Thirteen affected only Q or NQ cells, indicating significant differentiation of these cell types. doa4 strains, lacking ubiquitin hydrolase, affected viability and reproductive capacity in both cell types. More than 1300 mRNAs differentiating Q and NQ cell fractions were identified by microarray analysis. Gene-ontology analysis of Q-cell mRNAs showed significant increases in protein-encoding mRNAs involved in membrane maintenance, oxidative stress response, and signal transduction. NQ-cell mRNAs encode proteins involved in Ty-element transposition and DNA recombination, consistent with apoptosis in these cells. Consistent with preparation for rapid response to environmental stimuli, approximately 2000 protease-labile mRNAs were identified in Q cells. The differentiation of these cell types and the ability of genes to selectively affect the survival of Q or NQ cells in yeast are relevant to chronological aging, cell-cycle, genome-evolution, and stem-cell research and provides insight into complex responses that even simple organisms have to starvation. 10 replicates per cell-type. Channel 1 (Cy3) was the control of mixture of exponential and stationary phase cells. Channel 2 (Cy5) was the experimental cell-types of parental strain BY4742.
2010-05-27 | E-GEOD-8542 | biostudies-arrayexpress
Project description:This SuperSeries is composed of the following subset Series: GSE8542: BY4742 Quiescent and Non-quiescent replicates GSE8558: Proteinase K treatment of Q and NQ cells GSE8559: S288c Quiescent and Non-quiescent replicates GSE8560: Mutant1 GSE8561: Mutant2 Refer to individual Series
2010-05-29 | E-GEOD-8624 | biostudies-arrayexpress
Project description:The existence of two separate lineages of Escherichia coli O157:H7 has previously been reported, and research indicates that one of these lineages (lineage I) might be more pathogenic towards human hosts. We have previously shown that the more pathogenic lineage expresses higher levels of Shiga toxin 2 (Stx2) than the non-pathogenic lineage II. To evaluate why lineage 2 isolates do not express appreciable levels of toxin, two lineage 2 isolates (FRIK966 and FRIK2000) were chosen as representatives of lineage 2 and whole genome microarrays were performed using Agilent microarrays using the E. coli O157:H7 EDL933 lineage I clinical type isolate as a reference. Microarray results were utilized to evaluate what genes and pathways might be missing or differentially expressed. Quantitative RT-PCR was utilized to validate the microarray data. Based upon the transcriptome of Escherichia coli O157:H7 EDL933 an oligonucleotide microarray, made up of 60 mers was designed. A total of 4873 genes in an 8 x 15K Agilent microarray design. Designed using a custom script, specifications for gene specific oligos were based upon various design characteristics such as temperature of melting, 3’ location, specificity, lack of repeat nucleotides, etc. (Charbonnier et al., 2005). Arrays were manufactured using Agilent Sure-print technology. Each array consisted of duplicate elements for each gene randomly distributed with Agilent control elements included. All procedures were performed according to respective manufacturer protocols. Lineage I and lineage II strains were grown overnight as described, a total of 10e7 cells were washed twice in fresh media, normalized based upon optical density, inoculated into fresh media, incubated at 37oC shaking 120 x g for 3 hours and then suspended in RNAprotect bacteria reagent (Qiagen Inc., Valencia, CA.). Total RNA was extracted using RNeasy Bacteria Mini Kit (Qiagen Inc., Valencia, CA.) and trace amounts of DNA were removed using RNase-Free DNase Set (Qiagen Inc., Valencia, CA.). RNA was quantified using Nanodrop system (NanoDrop Technologies, Wilmington, DE) and quality confirmed by electrophoresis on a Bio-rad Experion system (BioRad XXX). For each sample, 10 ug of RNA were labeled with either CyDye3-dCTP or CyDye5-dCTP (Perkin Elmer) using the LabelStar kit (Qiagen Inc., Valencia, CA.) and Random nonamers (Integrated DNA Technologies ). Labeled cDNA were hybridized to the microarray using Agilent Hi-RPM hybridization solution in an Agilent Hybridization chamber (Agilent.). A total of 8 arrays, each with duplicate elements for each gene, alternating dye swap for each replicate (4 biological replicates), were analyzed to obtain genes that were consistently and differentially regulated in comparison to EDL933, while limiting false discover rate (FDR) below a stringent 5% (Benjamini and Hochberg, 1995).
2009-08-31 | E-GEOD-17070 | biostudies-arrayexpress
Project description:This series represents the effects of OVA, tg-IL-13, and direct effects of IL-13 on airway epithelial cells. Experiment Design: Type of experiment: comparison of three related murine models of asthma, 1) Ova model, 2) tg-IL-13 model, 3) IL-13/Epi model. Two control groups were used: PBS challenged mice to control for the effects of OVA and tg-IL-13+ and STAT6-/- mice as controls for tg-IL-13 and IL-13/Epi mice. Measurements were made in each model in two types of tissue samples, whole lung and tracheal perfusate. Experimental factors: Allergen vs sham challenge, natural STAT6 expression vs transgenic expression and no expression. The number of hybridizations performed in the experiment: 50 (25 whole lung and 25 tracheal perfusate). The type of reference used for the hybridizations, if any: Pooled reference from lung (whole lung or tracheal perfusate) from untreated wildtype mice. Hybridization design: two-color hybridizations with reference design. Each individual sample was hybridized to a separate array. Quality control steps taken: 5 experimental replicates per group. RNA integrity assessed by Agilent Bioanalyzer. Arrays with low signals, high background, or high spatial variation rejected and corresponding samples reanalyzed. URL of any supplemental websites or database accession numbers:GEO (http://www.ncbi.nlm.nih.gov/geo, accession number GSE1438). ********** Samples used, extract preparation and labeling: The origin of the biological sample: homogenized whole lung from mouse and perfusate of mouse trachea. Manipulation of biological samples and protocols used: Whole lungs mechanically homogenized in 7.0 ml Trizol reagent (Invitrogen) for total RNA collection. Tracheas perfused with lysis buffer (Qiagen Rneasy kit) for total RNA collection. Protocol for preparing the hybridization extract and labeling: Preparation of aminoallyl-UTP labeled whole lung cDNA was performed as described [1]. Total RNA from tracheal perfusate (1.0 - 1.5 g per sample) was amplified and labeled with aminoallyl-dUTP using the MessageAmp aRNA kit (Ambion). Labeled cRNAs were coupled to Cy3 or Cy5 dyes (CyScribe, Amersham Biosciences) and purified as described [1]. Labeling protocol(s): Coupled to Cy3 or Cy5.External controls (spikes): none. ********** Hybridization procedures and parameters: Hybridizations were performed as described [1] except Ambion SlideHyb Glass Array Hybridization Buffer #1 (neat concentration) was used as hybridization buffer and hybridizations were carried out for 40 h at 55 °C. Following hybridization, arrays were washed in 1X SSC with 0.03% SDS (wash 1), 0.2X SSC (wash 2), and 0.05X SSC (wash 3) for five minutes for each wash. ********** Measurement data and specifications: Arrays were scanned using an Axon Genepix 4000B scanner and GenePix Pro Analysis 5.0 software; laser power 100%, 10 micron resolution, PMT optimized for each array. Data from GPR files are available from GEO (http://www.ncbi.nlm.nih.gov/geo, accession number GSE1438).The âprint-tip loessâ normalization was used to correct for within-array dye and spatial effects and single channel quantile normalization was used to facilitate comparison between arrays. No background subtraction was performed. We used functions in the library marrayNorm of the R / Bioconductor package to perform these normalizations. After normalization we determined the log2 ratio of experimental sample intensity to reference sample intensity for each probe on each array. ********** Array Design: General array design Array design name: UCSF 10Mm Mouse v.2 Oligo Array (GEO GPL1089) and UCSF Gladstone 18K Mouse v.2 Oligo Array (GPL1196) Platform type: spotted oligonucleotides (70mers) Surface and coating specification: aminosilane coated glass slides Physical dimensions of array support (e.g. of slide): 25 x 75 x 1.0 mm Number of features on the array: GPL1089: 19152, GPL1196: 18240 (including empty and duplicate features) For production protocol, see http://www.microarrays.org/pdfs/PrintingArrays.pdf Feature information is available from GEO (GSE1438). Reporters: synthetic single stranded oligonucleotides. Sequence and annotation information available from GEO (GPL1089 and GPL1196) Method of reporter preparation: synthesized by Operon. The spotting protocols used, including the array substrate, the spotting buffer, and any post-printing processing, including cross-linking: see http://www.microarrays.org/pdfs/PrintingArrays.pdfAny additional treatment performed prior to hybridization: none. ********** Reference: 1. Barczak A, Rodriguez MW, Hanspers K, Koth LL, Tai YC, et al. (2003) Spotted long oligonucleotide arrays for human gene expression analysis. Genome Res 13: 1775-1785. Keywords = IL-13 Keywords = Epithelial Keywords = differential gene expression Keywords = microarray
2010-06-10 | E-GEOD-1438 | biostudies-arrayexpress
Project description:Anopheles gambiae mosquitoes exhibit an endophilic, nocturnal blood feeding behavior. Despite the importance of light as a regulator of malaria transmission, our knowledge on the molecular interactions between environmental cues, the circadian oscillators and the host seeking and feeding systems of the Anopheles mosquitoes is limited. In the present study, we show that the blood feeding behavior of mosquitoes is under circadian control and can be modulated by light pulses, both in a clock dependent and in an independent manner. Short light pulses (~2-5 min) in the dark phase can inhibit the blood-feeding propensity of mosquitoes momentarily in a clock independent manner, while longer durations of light stimulation (~1-2 h) can induce a phase advance in blood-feeding propensity in a clock dependent manner. The temporary feeding inhibition after short light pulses may reflect a masking effect of light, an unknown mechanism which is known to superimpose on the true circadian regulation. Nonetheless, the shorter light pulses resulted in the differential regulation of a variety of genes including those implicated in the circadian control, suggesting that light induced masking effects also involve clock components. Light pulses (both short and longer) also regulated genes implicated in feeding as well as different physiological processes like metabolism, transport, immunity and protease digestions. RNAi-mediated gene silencing assays of the light pulse regulated circadian factors timeless, cryptochrome and three takeout homologues significantly up-regulated the mosquito's blood-feeding propensity. In contrast, gene silencing of light pulse regulated olfactory factors down-regulated the mosquito's propensity to Our study suggests that the mosquitoâs feeding behavior is under circadian control. Long and short light pulses can induce inhibition of blood-feeding through circadian and unknown mechanisms, respectively, that involve chemosensory factors. A series of assays were performed to assess transcriptomic changes in mosquitoes upon light stimulation and blood feeding in order to assess relationships between photic stimulation and modulation of feeding behavior.
2010-06-24 | E-GEOD-13560 | biostudies-arrayexpress
Project description:Background: The mosquito Anopheles gambiae is a major vector of human malaria. Increasing evidence indicates that blood cells (hemocytes) comprise an essential arm of the mosquito innate immune response against both bacteria and malaria parasites. To further characterize the role of hemocytes in mosquito immunity, we undertook the first genome-wide transcriptomic analyses of adult female An. gambiae hemocytes following infection by two species of bacteria and a malaria parasite. Results: We identified 4047 genes expressed in hemocytes, using An. gambiae genome-wide microarrays. While 279 transcripts were significantly enriched in hemocytes relative to whole adult female mosquitoes, 959 transcripts exhibited immune challenge-related regulation. The global transcriptomic responses of hemocytes to challenge with different species of bacteria and/or different stages of malaria parasite infection revealed discrete, minimally overlapping, pathogen-specific signatures of infection-responsive gene expression; 105 of these represented putative immunity-related genes including anti-Plasmodium factors. Of particular interest was the specific co-regulation of various members of the Imd and JNK immune signaling pathways during malaria parasite invasion of the mosquito midgut epithelium. Conclusion: Our genome-wide transcriptomic analysis of adult mosquito hemocytes reveals pathogen-specific signatures of gene regulation and identifies several novel candidate genes for future functional studies. In order to identify hemocyte-specific and immune-responsive transcripts, we first compared transcripts expressed in hemocytes from one day old sugar-fed mosquitoes to transcripts detected in whole mosquitoes of the same age and feeding status. This resulted in identification of the hemocyte-enriched transcriptome. We then compared hemocytes from 1 day old mosquitoes, 1 hour after immune challenge with heat-killed Escherichia coli or Micrococcus luteus, to control female mosquitoes injected with sterile PBS to determine the bacteria challenge responsive transcriptomes. We used heat-killed bacteria in these assays, because our primary interest was in identifying the bacterial responsive transcriptome and to avoid the potentially confounding effects of altered gene expression due to the lethal effects of a systemic infection associated with injection of living bacteria. Lastly, we compared hemocytes from mosquitoes at 24 hours and 19 days after ingestion of a blood meal infected with Plasmodium berghei to mosquitoes of the same age fed a non-infected blood meal to determine the ookinete and sporozoite infection responsive transcriptomes, respectively. This design resulted in a total of five experimental treatments. The following samples are not included in this submission: Hemo E coli vs. hemo unchallenged A Hemo E coli vs. hemo unchallenged B Hemo m luteus vs. hemo unchallenged A Hemo m luteus vs. hemo unchallenged B
2011-10-31 | E-GEOD-33269 | biostudies-arrayexpress
Project description:The local background was subtracted from the fluorescent value of each spot. Feature intensities were extracted from scanned microarray images using GenePix Pro 5.1. (Axon Instruments). The images were visually inspected, and spots from low-quality areas of the array were flagged and excluded from further analysis. Spots were also excluded from analysis if both the combined fluorescent intensity for both channels was less than 1.4 times that of the local background and the pixel-by-pixel correlation coefficient of the spot was less than 0.4. The fluorescence ratios were normalized as previously described (Turton and Gant oncogene 2001website). A hierarchical clustering algorithm (M. Eisen Brown and Botstein PNAS 98 website) was applied to those genes that fulfilled all of the following conditions: they were not flagged; they were differentially expressed (equal or higher than 1.5 fold upregulated or equal or lower than 2 fold downregulated); they had p-values equal or less than 0.02; and they were consistent on at least 4 out of the 5 arrays.
2010-05-27 | E-GEOD-2069 | biostudies-arrayexpress