Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of S. cerevisiae GPCR mutants gpr1 and gpa2

ABSTRACT: Experiment design,Type of experiment: Gene expression profiling analysis of S.cerevisiae W303-1a.,Experimental factor: genetic variation (WT, gpr1 and gpa2 mutant strains).,Number of hybridizations: 6. Single-colour labeling was performed for each sample.,Sample preparation:,Total RNA was prepared using FastRNA Pro Red Kit (BIO 101 Systems) according to the manufacturers' instructions.,Labeling protocol,10 Âµg of total RNA was reverse transcribed using the SuperScript Choice system for cDNA synthesis (Life Technologies) according to the protocol recommended by Affymetrix. The oligonucleotide used for priming was 5â-ggccagtgaattgtaatacgactcactatagggaggcgg-(t)24-3â (Genset Oligo). Double-stranded cDNA was cleaned by phenol:chloroform extraction and the aqueous phase removed by centrifugation through Phase-lock Gel (Eppendorf). In vitro transcription was performed on 1 Âµg of cDNA using the Enzo BioArray High Yield RNA transcript labelling kit (Enzo Diagnostics). The cRNA was cleaned using RNAeasy clean-up columns (Qiagen). The cRNA was fragmented by heating in 40 mM Tris-acetate pH 8.1, 100 mM KOAc, 30 mM MgOAc.,Hybridization protocol,10 Âµg of fragmented cRNA were hybridised (45Â°C, 16 hours). Hybridization was controled by use of the GeneChipâ¢ Eukaryotic Hybridization Control Kit (Affymetrix). Washing and staining was performed in a Fluidics Station 400 (Affymetrix) using the protocol EukGE-WS2v4.,Measurement data, specifications and data processing,Scanning was performed in an Affymetrix GeneChip scanner. Chip analysis was performed using the Affymetrix Microarray Suite v5.,Changes in gene expression were assessed by looking for concordant changes between all replicates using a signed Wilcoxon rank test. The âchangeâ p-value threshold was < 0.025 for increase and > 0.975 for decrease.,Array design,Microarray analysis was performed using YGS98 GeneChipsâ¢ oligonucleotide arrays (Affymetrix #900256) representing approximately 6000 ORFs. Sequences and GenBank accession numbers of all probe sets are available at the Affymetrix home page http://www.affymetrix.com/index.affx.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Katsuhiko Shirahige

PROVIDER: E-GEOD-970 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Glucose-dependent cell size is regulated by a G protein-coupled receptor system in yeast Saccharomyces cerevisiae.

Tamaki Hisanori H Yun Cheol-Won CW Mizutani Tomohiro T Tsuzuki Takahiro T Takagi Yukinobu Y Shinozaki Makiko M Kodama Yukiko Y Shirahige Katsuhiko K Kumagai Hidehiko H

Genes to cells : devoted to molecular & cellular mechanisms 20050301 3

In the yeast, Saccharomyces cerevisiae, cell size is affected by the kind of carbon source in the medium. Here, we present evidence that the Gpr1 receptor and Gpa2 Galpha subunit are required for both maintenance and modulation of cell size in response to glucose. In the presence of glucose, mutants lacking GPR1 or GPA2 gene showed smaller cells than the wild-type strain. Physiological studies revealed that protein synthesis rate was reduced in the mutant strains indicating that reduced growth r ...[more]

PMID: 15743410

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Dataset Information

Transcription profiling of S. cerevisiae GPCR mutants gpr1 and gpa2

Publications

Glucose-dependent cell size is regulated by a G protein-coupled receptor system in yeast Saccharomyces cerevisiae.

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