Identification of genes regulated by acute overexpression of Yox1 and Yhp1 in S. cerevisiae
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ABSTRACT: We report gene expression data for strains that overexpress the transcriptional repressors Yox1 or Yhp1 from a synthetic, estradiol-inducible promoter. Gene expression was also measured in strains that overexpress unphosphorylatable alleles of either Yox1 or Yhp1, to determine whether blocking phosphorylation of these proteins by Cdk1 changes the spectrum of regulated genes. Finally, gene expression was measured in corresponding control strains in which Yox1/Yhp1 proteins were expressed from their endogenous promoters. These data demontsrate that Yox1 is a stronger repressor than Yhp1, although they regulate the same genes. In addition, phosphodeficient alleles of each TF regulate the same genes as the wild type proteins.
Project description:We report gene expression data for yeast strains expressing mutated versions of the transcriptional repressors Yox1 and Yhp1 that can't be phosphorylated by Cdk1. For comparision, gene expression was also measured in cells with deletions in one or both repressors. Genes that are downregulatd in phosphomutant strains and upregulated in deletion strains are likely to be target genes of these transcriptional repressors.
Project description:Transcription profiles across the cell cycle of yox1 deleted yeast cells. BY2399 cells (yox1 delete cells isogenic with W303a) were arrested with alpha factor in YEPD media. After release from arrest cells were sampled every 5 min for 2 hr. Keywords: cell cycle time course
Project description:Transcription profiling of fission yeast /yox1/ deletion and genome wide location analysis of Yox1p and Cdc10p transcription factors reveal a negative feedback interaction: /yox1 /is transcriptionally activated by MBF, and Yox1p in turn transcriptionally represses the MBF target genes (including /yox1 /itself).
Project description:Transcription profiling of fission yeast /yox1/ deletion and genome wide location analysis of Yox1p and Cdc10p transcription factors reveal a negative feedback interaction: /yox1 /is transcriptionally activated by MBF, and Yox1p in turn transcriptionally represses the MBF target genes (including /yox1 /itself).
Project description:Transcription profiles across the cell cycle of yox1 deleted yeast cells. BY2399 cells (yox1 delete cells isogenic with W303a) were arrested with alpha factor in YEPD media. After release from arrest cells were sampled every 5 min for 2 hr. Keywords: cell cycle time course Cells were synchronized with alpha factor and sampled every 5 min across 2 cell cycles. A total of 25 samples were analyzed. Technical replicates were included. cDNA of the cell cycle samples were labelled with Cy5. For Cy3 labelling, asynchronous yeast population was used.
Project description:mRNA processing bodies (P-bodies) are cytoplasmic granules that form in eukaryotic cells in response to numerous stresses to serve as sites of degradation and storage of mRNAs. Functional P-bodies are critical for the DNA replication stress response in yeast, yet the repertoire of P-body targets and the mechanisms by which P-bodies promote replication stress resistance are unknown. In this study, we identify the complete complement of mRNA targets of P-bodies during replication stress induced by hydroxyurea treatment. The key P-body protein Lsm1 controls the abundance of HHT1, ACF4, ARL3, TMA16, RRS1 and YOX1 mRNAs to prevent their toxic accumulation during replication stress. Accumulation of YOX1 mRNA causes aberrant down-regulation of a network of genes critical for DNA replication stress resistance. Among these genes we identify ALD6, whose de-repression is critical to prevent accumulation of acetaldehyde, a strongly toxic molecule during DNA replication stress.
Project description:Two homeodomain proteins, Yox1 and Yhp1, act as repressors at early cell cycle boxes (ECBs) to restrict their activity to the M/G1 phase of the cell cycle in budding yeast. These proteins bind to Mcm1 and to a typical homeodomain binding site. The expression of Yox1 is periodic and directly correlated with its binding to, and repression of, ECB activity. The absence of Yox1 and Yhp1 or the constitutive expression of Yox1 leads to the loss of cell-cycle regulation of ECB activity. Therefore, the cell-cycle-regulated expression of these repressors defines the interval of ECB-dependent transcription. Twenty-eight genes, including MCM2-7, CDC6, SWI4, CLN3, and a number of genes required during late M phase have been identified that are coordinately regulated by this pathway. Keywords: cell cycle time course
Project description:Analysis of gene expression across the cell cycle from wild type cells, and cells expressing alleles of Yox1, Yhp1, Hcm1, and Tos4 that cannot be phosphorylated by Cdk1. Expression of S-phase and M/G1 transcripts are downregulated when phosphorylation of these factors is blocked, demonstrating that Cdk1 promotes expression of late cell cycle genes.