Regulation of S-phase transcription factors by Cdk1
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ABSTRACT:
ORGANISM(S): Saccharomyces cerevisiae
PROVIDER: GSE55121 | GEO | 2014/04/10
SECONDARY ACCESSION(S): PRJNA238581
REPOSITORIES: GEO
Similar Datasets
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. These experiments are two-color hybridizations of RNA isolated from synchronized wild type (WT) or phosphomutant (4P) cells, compared to RNA from asyncrhonous wild type cells in mid-log phase. Wild type and mutant cells were synchronized in G1 phase, released into the cell cycle and samples collected at 15 minute intervals. Each time course was carried out in duplicate, the replicate experiment was performed as a dye swap.
2014-04-10 | E-GEOD-55121 | biostudies-arrayexpress
Project description: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.
2022-07-25 | GSE188179 | GEO
Project description:We addressed the role of cell cycle regulator CDK1 during differentiation by either inhibiting its activity with the specific inhibitor RO3306 (10µM) or by knocking its expression down using specific shRNAs, during mesendoderm differentiation. Human pluripotent stem cells (H9 cells) were synchronized in their cell cycle and then induced to differentiate into mesendoderm for 24h. shRNAs against CDK1 were induced 4 days before starting the differentiation. The CDK1 inhibitor was added to the medium 8h after the induction of differentiation. At the end of the experiment, the cells were harvested and processed for total RNA extraction. RNA-IP was carried out using RiboCluster Profiler RIP-Assay Kit (MBL) following the manufacture’s recommendations.
2021-05-31 | GSE130827 | GEO
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.
2022-07-25 | GSE188177 | GEO
Project description:The retinoblastoma tumor suppressor protein (Rb) regulates early G1 phase checkpoints, including the DNA damage response, as well as cell cycle exit and differentiation. The widely accepted model of G1 cell cycle progression proposes that cyclin D:Cdk4/6 partially inactivates the Rb tumor suppressor during early G1 phase by progressive multi-phosphorylation, termed hypo-phosphorylation, resulting in release of E2F transcription factors. However, this model remains largely unproven biochemically and the biologically active form(s) of Rb remains unknown. Here we find that Rb is un-phosphorylated in G0 cells and becomes exclusively mono-phosphorylated throughout all of early G1 phase by cyclin D:Cdk4/6. Early G1 phase mono-phosphorylated Rb is composed of 14 independent isoforms that are all targeted by the E1a oncoprotein, but each shows a preferential binding pattern to specific E2F1-4 transcription factors. At the late G1 Restriction Point, cyclin E:Cdk2 inactivates Rb by a quantum hyper-phosphorylation (>12 phosphates/Rb). Cells undergoing a DNA damage response activate cyclin D:Cdk4/6 to generate mono-phosphorylated Rb that regulates global transcription. In contrast, a non-phosphorylatable ?Cdk-Rb allele was non-functional for regulating a DNA damage response, but functional for driving cell cycle exit and differentiation during myogenesis. These observations fundamentally change our understanding of G1 cell cycle progression and show that there is no progressive multi-phosphorylation or hypo-phosphorylation inactivation of Rb during early G1 phase by cyclin D:Cdk4/6. Instead, cyclin D:Cdk4/6 generates functionally active, mono-phosphorylated Rb that is the only Rb isoform present in cells during early G1 phase.
2014-04-03 | GSE56453 | GEO
Project description:The retinoblastoma tumor suppressor protein (Rb) regulates early G1 phase checkpoints, including the DNA damage response, as well as cell cycle exit and differentiation. The widely accepted model of G1 cell cycle progression proposes that cyclin D:Cdk4/6 partially inactivates the Rb tumor suppressor during early G1 phase by progressive multi-phosphorylation, termed hypo-phosphorylation, resulting in release of E2F transcription factors. However, this model remains largely unproven biochemically and the biologically active form(s) of Rb remains unknown. Here we find that Rb is un-phosphorylated in G0 cells and becomes exclusively mono-phosphorylated throughout all of early G1 phase by cyclin D:Cdk4/6. Early G1 phase mono-phosphorylated Rb is composed of 14 independent isoforms that are all targeted by the E1a oncoprotein, but each shows a preferential binding pattern to specific E2F1-4 transcription factors. At the late G1 Restriction Point, cyclin E:Cdk2 inactivates Rb by a quantum hyper-phosphorylation (>12 phosphates/Rb). Cells undergoing a DNA damage response activate cyclin D:Cdk4/6 to generate mono-phosphorylated Rb that regulates global transcription. In contrast, a non-phosphorylatable ?Cdk-Rb allele was non-functional for regulating a DNA damage response, but functional for driving cell cycle exit and differentiation during myogenesis. These observations fundamentally change our understanding of G1 cell cycle progression and show that there is no progressive multi-phosphorylation or hypo-phosphorylation inactivation of Rb during early G1 phase by cyclin D:Cdk4/6. Instead, cyclin D:Cdk4/6 generates functionally active, mono-phosphorylated Rb that is the only Rb isoform present in cells during early G1 phase. Global transcriptional analysis of murine embryonic fibroblasts (MEFs) with conditional deletion of the endogenous RB gene by treatment with cell permeable TAT-Cre. Comparison to unaltered MEFs and MEFs with physiological level of exogenous wildtype or phospho-mutant RB expressed at time of RB gene deletion.
2014-04-03 | E-GEOD-56453 | biostudies-arrayexpress
Project description:Effect of Cell cycle regulator CDK1 on Cell Differentiation (RNA-IP CDK1 inh KD)
| PRJNA541529 | ENA
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
2006-08-31 | GSE5283 | GEO
Project description:Purpose: Determine whether Cdk1 only regulates a specific subset of tDNAs, or whether Cdk1 has a more global impact on tDNA expression; Method: We study the expression of tRNA genes in WT cells and cdk1-as1 mutants during S phase by small-RNAseq; Result: We found that expression of at least 77% of all tDNAs is significantly downregulated in S-phase in the cdk1-as1 mutant compared with the WT strain; Conclussions: Cdk1 promotes transcription of the majority of tRNA genes during the cell cycle.
2018-09-10 | GSE116224 | GEO
Project description:Entry of mammalian cells into DNA synthesis (S-phase) represents a key event in cell division. According to the current cell cycle models, the Cdc7 kinase represents the essential and rate-limiting trigger of DNA replication, acting together with the cyclin-dependent kinase Cdk2. Using chemical genetic systems which allow an acute shutdown of Cdc7 in in vitro cultured cells as well as in vivo in a living mouse, that Cdc7 is dispensable for cell division of many different cell types. Proteomic and phosphoproteomic analysis were performed to elucidate the compensating mechanism. We demonstrate that Cdk1-cyclin B, a kinase currently thought to act exclusively during mitosis, is also active during G1/S transition both in cycling cells and in cells exiting quiescence. We show that Cdc7 and Cdk1-cyclin B play redundant roles during G1/S transition, and at least one of these kinases is needed to allow S-phase entry. These observations revise our fundamental understanding of cell cycle progression by demonstrating that Cdk1-cyclin B physiologically regulates two distinct transitions during cell division cycle, while Cdc7 plays a redundant function in DNA replication. In TMT1, to analyze the expression of Cdc7 and its partners upon auxin-mediated induction of Cdc7 degradation, we performed a TMT analysis of ES cells treated with IAA for 24 hours. In TMT2, to analyze the phosphorylation of Cdc7 targets as a function of time upon degradation of the kinase we performed the time-resolved TMT analysis of ES cells subjected to Cdc7 degradation for three different amounts of time. In TMT3, to determine the ability of Cdk1 to phosphorylate Mcm2, we analyzed the phosphoproteome of mES cells and focused on the phosphorylation of Mcm2 upon Cdk1 inhibition.
2022-03-11 | PXD025625 | Pride