Determination of transcriptional rates and mRNA levels during osmotic stress in S. cerevisiae wild type and cbc1/sto1 mutant cells using Genomic Run On (GRO)

We analyzed the effect of the elimination of the cap-binding protein Cbc1 on transcription rate and mRNA amount during the response to osmotic stress using the Genomic Run-on assay This study focus on the transcriptional activity of RNA pol II in yeast genes during stress response. S. cerevisiae cells grown in YPD to exponential phase were subjected to osmotic stress and then processed for Genomic Run-On. Home-made microarrays containing entire ORFs were used. Three replicates of each time point were done. Each replicate has been hybridized on a different macroarray. A single DNA genomic hybridization was done on the same microarrays for normalization. Data were normalized by Median Absolute Deviation (MAD) using the ArrayStat software.