Fhl1 and lfh1 ChIP-chip
Ontology highlight
ABSTRACT:
ORGANISM(S): Saccharomyces cerevisiae
PROVIDER: GSE1930 | GEO | 2004/11/09
SECONDARY ACCESSION(S): PRJNA90961
REPOSITORIES: GEO
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Project description:Fhl1-9myc ChIP-chip, YPD, OD600=0.8, 2 arrays with duplicate spotting of yeast intergenic regions. AND Ifh1-9myc ChIP-chip, cells grown in YPD, OD600=0.8, 2 arrays with duplicate spotting of yeast intergenic regions. Keywords = Fhl1
2010-06-05 | E-GEOD-1930 | biostudies-arrayexpress
Project description:Affymetrix experiment performed on RNA isolated from Wild type, FHL1 deleted and FHL1,IFH1 double deleted strains. Data aquired in duplicate. In S. cerevisiae the mRNAs from the 138 ribosomal protein (RP) genes are amongst the most abundant in the cell, and their transcription is regulated tightly so that they are the most prominent cluster in most transcriptome experiments. It has recently been observed that the proteins Fhl1p and Ifh1p are found almost exclusively at RP genes (Lee et al., Science, 298, 799-804, 2002;Jorgensen et al., Genes Dev, 18, 2491-2505 2004; Schawalder et al Nature in press; Rudra et al , EMBO J. submitted), and data suggests they are the true transcription factors. This experiment utilizes Affymetrix arrays to measure the level of all mRNAs in cells with a deletion of FHL1 or of both FHL1 and IFH1, compared to the wild type strain, W303. Such mutant cells are viable but grow very slowly (Hermann-Le Denmat S. et al., Mol Cell Biol, 14, 2905-2913, 1994; Cherel and Thuriaux, Yeast, 11, 261-270, 1995). Total RNA was made from log phase cells, amplified according to conventional methods, and hybridized to the array. (Since the total RNA/cell of the mutants is only a fifth of the wt, each mutant required five times the number of cells to provide the same amount of RNA.) The experiment was carried out in duplicate and the six tables provide the raw data from the wt, W303, delFHL1, and delFHL1, delIFH1 strains. Although we expected that the RP genes would be substantially reduced in these mutant cells, they are only slightly less abundant than normal in comparison to all other mRNAs! The interesting result is that the cells respond to a severe deficiency of ribosomes by reducing the level of nearly all mRNAs to match the capacity of the translational apparatus. Keywords = Ribosome Keywords = FHL1 Keywords = IFH1. Keywords: parallel sample
2004-12-22 | GSE2096 | GEO
Project description:Array design -Platform: amino-silane coated glass slides (GAPS II, Corning) -S. cerevisiae intergenic regions amplified from S288C genomic DNA (ResGen) using the intergenic region primer oligonucleotides (ResGen) (Harismendy et al. EMBO J. 22(18): 4738-4747, 2003). The primers allow the amplification of the sequence located on either side of elements such as open reading frames, tRNAs, small nuclear RNAs, Ty elements, solo δ, etc. A complete description of the primers can be obtained on the web site (ftp://ftp.resgen.com/pub/genepairs/yeast_intergenic). -The PCR products were purified and spotted using an automated arrayer (MicroGrid II, BioRobotics) Experiment design -Type of experiment: genome wide binding analysis -Experimental factor: genotype (Wild-Type, Fhl1-myc, Ifh1-myc); drug (rapamycin) -Number of hybridization performed in the experiment: 15 (four conditions with three to five biological replicates each) -Reference used for each condition: ChIP from the wild-type strain -Quality control: one dye-swap per condition Samples used, extract preparation and labeling -The origin of the biological sample: o Species: S. cerevisiae o Cell type: (1) W303; (2) W303+IFH1-13myc-TRP1MX6;(3) W303+FHL1-13myc-HIS3MX6. -Growth conditions: log phase (2x107/mL) at 30ºC in YPD medium; when indicated rapamycin was added at the final concentration of 200 ng/mL for 90 min. -Cross-linking for 30 min at 30ºC -Chromatin immunoprecipitation: Dynabeads M280 coupled to sheep anti-mouse IgG or sheep anti-rabbit IgG and mouse monoclonal antibodies against the myc epitope (culture supernatant of 9E10) were used for immunoprecipitation according to standard techniques. -Labeling protocol(s): each IP was blunt-ended, ligated to linker DNA, then subjected to Ligation Mediated (LM)-PCR in the presence of amino-allyl dUTP. Labeling was performed for 1 hour at room temperature in the presence of NHS-ester Cy3 or Cy5 in 0.1 M Sodium Carbonate. The labeling reaction was stopped with 2 M Hydroxylamine and the labeled DNA purified. Hybridization procedures and parameters -Standard protocols were used Measurement data and specifications -Software for scanning: GenePix Pro 4.0 (Axon Instruments, Inc.) -Scanner: GenePix 4000A (Axon Instruments, Inc.) Keywords: dose response
2004-11-10 | GSE1944 | GEO
Project description:Forkhead transcription factor FHL1 gene was required for replicative lifespan as well as cell proliferation in yeast. In this study, to see how Fhl1p determines the lifespan, we performed a DNA microarray analysis of heterozygous diploid strain deleted for FHL1 and, from the Fhl1p-target genes, screened for a lifespan-related gene. Transcriptomic profiles showed large increases and moderate decreases in transcripts in the fhl1/FHL1 mutant. Pathway analysis suggested upregulation of most of carbon metabolisms (glycolysis, gluconeogenesis, and TCA cycle) and downregulation of translation (ribosomal proteins and translation factors) in the FHL1-deficient mutant. We found a remarkable downregulation of a set of phosphate metabolism genes and ribonucleotide reductase large subunit genes, RNR1 and RNR3. We demonstrated that Fhl1p regulates RNR1 gene transcription to maintain dNTP levels, modulating longevity against replication stress and genomic instability.
2017-03-02 | GSE95591 | GEO
Project description:Array design -Platform: amino-silane coated glass slides (GAPS II, Corning) -S. cerevisiae intergenic regions amplified from S288C genomic DNA (ResGen) using the intergenic region primer oligonucleotides (ResGen) (Harismendy et al. EMBO J. 22(18): 4738-4747, 2003). The primers allow the amplification of the sequence located on either side of elements such as open reading frames, tRNAs, small nuclear RNAs, Ty elements, solo δ, etc. A complete description of the primers can be obtained on the web site (ftp://ftp.resgen.com/pub/genepairs/yeast_intergenic). -The PCR products were purified and spotted using an automated arrayer (MicroGrid II, BioRobotics) Experiment design -Type of experiment: genome wide binding analysis -Experimental factor: genotype (Wild-Type, Fhl1-myc, Ifh1-myc); drug (rapamycin) -Number of hybridization performed in the experiment: 15 (four conditions with three to five biological replicates each) -Reference used for each condition: ChIP from the wild-type strain -Quality control: one dye-swap per condition Samples used, extract preparation and labeling -The origin of the biological sample: o Species: S. cerevisiae o Cell type: (1) W303; (2) W303+IFH1-13myc-TRP1MX6;(3) W303+FHL1-13myc-HIS3MX6. -Growth conditions: log phase (2x107/mL) at 30ºC in YPD medium; when indicated rapamycin was added at the final concentration of 200 ng/mL for 90 min. -Cross-linking for 30 min at 30ºC -Chromatin immunoprecipitation: Dynabeads M280 coupled to sheep anti-mouse IgG or sheep anti-rabbit IgG and mouse monoclonal antibodies against the myc epitope (culture supernatant of 9E10) were used for immunoprecipitation according to standard techniques. -Labeling protocol(s): each IP was blunt-ended, ligated to linker DNA, then subjected to Ligation Mediated (LM)-PCR in the presence of amino-allyl dUTP. Labeling was performed for 1 hour at room temperature in the presence of NHS-ester Cy3 or Cy5 in 0.1 M Sodium Carbonate. The labeling reaction was stopped with 2 M Hydroxylamine and the labeled DNA purified. Hybridization procedures and parameters -Standard protocols were used Measurement data and specifications -Software for scanning: GenePix Pro 4.0 (Axon Instruments, Inc.) -Scanner: GenePix 4000A (Axon Instruments, Inc.) Keywords: dose response
2010-06-05 | E-GEOD-1944 | biostudies-arrayexpress
Project description:Affymetrix experiment performed on RNA isolated from Wild type, FHL1 deleted and FHL1,IFH1 double deleted strains. Data aquired in duplicate.,In S. cerevisiae the mRNAs from the 138 ribosomal protein (RP) genes are amongst the most abundant in the cell, and their transcription is regulated tightly so that they are the most prominent cluster in most transcriptome experiments. It has recently been observed that the proteins Fhl1p and Ifh1p are found almost exclusively at RP genes (Lee et al., Science, 298, 799-804, 2002;Jorgensen et al., Genes Dev, 18, 2491-2505 2004; Schawalder et al Nature in press; Rudra et al , EMBO J. submitted), and data suggests they are the true transcription factors.,This experiment utilizes Affymetrix arrays to measure the level of all mRNAs in cells with a deletion of FHL1 or of both FHL1 and IFH1, compared to the wild type strain, W303. Such mutant cells are viable but grow very slowly (Hermann-Le Denmat S. et al., Mol Cell Biol, 14, 2905-2913, 1994; Cherel and Thuriaux, Yeast, 11, 261-270, 1995). Total RNA was made from log phase cells, amplified according to conventional methods, and hybridized to the array. (Since the total RNA/cell of the mutants is only a fifth of the wt, each mutant required five times the number of cells to provide the same amount of RNA.) The experiment was carried out in duplicate and the six tables provide the raw data from the wt, W303, delFHL1, and delFHL1, delIFH1 strains.,Although we expected that the RP genes would be substantially reduced in these mutant cells, they are only slightly less abundant than normal in comparison to all other mRNAs! The interesting result is that the cells respond to a severe deficiency of ribosomes by reducing the level of nearly all mRNAs to match the capacity of the translational apparatus.
2007-07-07 | E-GEOD-2096 | biostudies-arrayexpress
Project description:The dataset contains ChIP-Seq data of the Set3 and Hos2 proteins in Candida albicans, assayed in two morphological phases (yeast and hypha). The Set3 and Hos2 proteins in the respective strains carry 9myc epitopes and ChIP was performed with an anti-myc antibody. Included samples are the following: 1 input and 1 ChIP sample of an untagged wild type strain as negative control assayed in the yeast phase, 1 input and 3 ChIP biological replicates of the Set3-9myc strain in the yeast phase, 1 input and 2 ChIP biological replicates of the Set3-9myc strain in the hypha phase, 1 input and 2 ChIP biological replicates of the Hos2-9myc strain in the yeast phase, 1 input and 2 ChIP biological replicates of the Hos2-9myc strain in the hypha phase, 1 input and 3 ChIP biological replicates of Set3-9myc in a set1delta/delta background in the yeast phase. ChIP-Seq was performed of Candida albicans strains in two morphological phases (yeast and hypha). Yeast-phase cells were grown to the exponential phase in YPD at 30C. Hyphal differentiation was induced by resuspending the cells in YPD+20% Fetal Calf Serum and a shift of the growth temperature to 37C. Induction was performed for 30 minutes. Cells were crosslinked with 1% formaldehyde for 15 minutes at room temperature.
2013-01-01 | E-GEOD-38425 | biostudies-arrayexpress
Project description:The dataset contains ChIP-Seq data of the Set3 and Hos2 proteins in Candida albicans, assayed in two morphological phases (yeast and hypha). The Set3 and Hos2 proteins in the respective strains carry 9myc epitopes and ChIP was performed with an anti-myc antibody. Included samples are the following: 1 input and 1 ChIP sample of an untagged wild type strain as negative control assayed in the yeast phase, 1 input and 3 ChIP biological replicates of the Set3-9myc strain in the yeast phase, 1 input and 2 ChIP biological replicates of the Set3-9myc strain in the hypha phase, 1 input and 2 ChIP biological replicates of the Hos2-9myc strain in the yeast phase, 1 input and 2 ChIP biological replicates of the Hos2-9myc strain in the hypha phase, 1 input and 3 ChIP biological replicates of Set3-9myc in a set1delta/delta background in the yeast phase.
2013-01-01 | GSE38425 | GEO
Project description:C. albicans (strain CAI4-CIp10) was grown according to CRISP SOP. An overnight culture was started from a single colony in YPD-Tris,(100mM Tris.HCl) pH 7.4 and incubated overnight at 30C in shaking incubator. The next day, 500 ul culture was inoculated into 50 ml YPD-Tris, pH 7.4. The following day a fresh culture was inoculated in YPD-Tris to an OD600 of 0.2 and grown to OD600 of 0.8 at 30C in a shaking incubator. At this point the culture was split, diluted back to OD600 of 0.2 in fresh medium and stress agents added at time =0 (XS = 5mM H2O2, OS = 1M NaCl or a combination, OSXS). After 10 min the cultures were harvested by centrifugation and cell pellets flash frozen in liquid nitrogen. Three independent cultures were grown for RNA extraction for each isolate at each condition.<br><br>The control sample was obtained from cells with no stress agents added, harvested at time=0. RNA was extracted and transcript profiling carried out by microarray analysis using custom microarrays (Eurogentec).
2014-07-01 | E-MEXP-3003 | biostudies-arrayexpress
Project description:Transcriptional profiling of mid-log phase (OD600 ~ 0.8) E. coli BW25113 cells grown treated with 2 mM NaN3 for 10 minutes
2017-08-08 | GSE102322 | GEO