Project description:Genomic DNA from Saccharomyces cerevisiae W303-1A was isolated along with nucleosomal DNA from W303-1A and the yeast strain GAL:orc2-1. Nucleosomal DNA was isolated following a 2h release from a nocodazole block in which the medium was changed from galactose (YPAG) to glucose (YPAD).
Project description:The accumulation of unfolded proteins in the lumen of the endoplasmic reticulum (ER) causes stress and induces the unfolded protein response (UPR) which is characterised in part by the transcriptional induction of genes involved in assisting protein folding. Translational responses to ER stress have been less well described and here we report on a genome-wide analysis of translational regulation in the response to the ER stress-inducing agent dithiothreitol (DTT) in Saccharomyces cerevisiae. Although the observed polysome profiles were similar under control and ER stress conditions microarray analysis identified transcipt-specific translational regulation. Genes with functions in ribosomal biogenesis and assembly were translationally repressed under ER stress. In contrast mRNAs for known UPR genes, including the UPR transcription factor HAC1, the ER-oxidoreductase ERO1 and the ER-associated protein degradation (ERAD) gene DER1 were enriched in polysomal fractions under ER stress conditions. In addition, we show that splicing of HAC1 mRNA is required for efficient ribosomal loading and that Gcn2p is required for normal HAC1 splicing, so shedding light on the role of this protein kinase in the UPR pathway. Experiment Overall Design: Polyribosomes were extracted from S. cerevisiae cells treated with 2 mM DTT or water (control), and fractionated according to ribosome loading. Following RNA purification from these fractions, for each sub-polysomal and polysomal RNA sample, fractions from three independent extracts per treatment (DTT/control) were pooled.
Project description:Comparative transcriptome analysis of E. faecium E1162 and its isogenic mdxR deletion mutant during growth in BHI in mid-exponential phase (A660 = 0.3)
Project description:This study newly identified Tripelennamine (TA) as an inhibitor of yeast meiosis and sporulation. To examine if and how exposure of sporulating yeast cells to TA changes the meiotic transcriptional program cells were sporulated for 0, 4, and 8 hours in the presence or absence of 100 uM TA.
Project description:This is a polysome/monosome profiling study on yeast undergoing glucose depletion. Polysomes and monosomes are extracted and the transcripts bound to each fraction are measured by microarray. The study reveals that glucose depletion inhibits translation initiation in a mechanism involving eIF4A loss and 48S pre-initiation complex accumulation, while the pentose phosphate pathway is co-ordinately up-regulated
Project description:Comparative transcriptome analysis of E. faecium E1162 and its isogenic mdxR deletion mutant during growth in M1 with maltose in mid-exponential phase (A660 = 0.3)