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HU treated cells vs wildtype untreated cells (asynchronized cells)

ABSTRACT: DNA replication checkpiont kinase rad3 (human ATM/ATR-like kinase) and cds1 (human CHK2-like kinase) are known to restore stalled replication forks and prevent subsequent cell cycle progression during a replication block imposed by HU-treatment in fission yeast. In order to identify cell cycle-specific exression which are modulated by replication checkpoint kinase rad3 and cds1, microarray approach was used to profile asynchronous and synchronous cells of wildtype and mutants treated and untreated with HU Keywords: HU treated cells vs wildtype untreated cells

ORGANISM(S): Schizosaccharomyces pombe

PROVIDER: GSE4280 | GEO | 2007/12/20

SECONDARY ACCESSION(S): PRJNA104617

REPOSITORIES: GEO

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HU treated cells vs wildtype untreated cells (elutriation centrifugation synchronized cells)

Project description:DNA replication checkpiont kinase rad3 (human ATM/ATR-like kinase) and cds1 (human CHK2-like kinase) are known to restore stalled replication forks and prevent subsequent cell cycle progression during a replication block imposed by HU-treatment in fission yeast. In order to identify cell cycle-specific exression which are modulated by replication checkpoint kinase rad3 and cds1, microarray approach was used to profile asynchronous and synchronous cells of wildtype and mutants treated and untreated with HU Keywords: HU treated cells vs wildtype untreated cells

2007-12-20 | GSE4283 | GEO

Genome-wide Profiling of DNA Replication Timing in pombe.

Project description:DNA replication is initiated at multiple sites or origins enriched with AT-rich sequences at various times during the S-phase. While current studies of genome-wide DNA replication profiles have focused on the timing of replication and the location of origins, the efficiency of replication/firing at various origins remains unclear. In this study, we show different efficiencies of DNA replication at various loci by using ORF-specific DNA microarrays. DNA copy-number increases as a function of time at individual loci are approximated to near-sigmoidal models for estimation of replication initiation and completion timings in HU-challenged cells. Duplicating times (from initiation to completion) vary from loci to loci, partly contributing to various firing efficiencies at origins. DNA replication timing profiles are strikingly similar to the reported patterns of enriched ssDNA, suggesting that majority stalled forks are restored for resumption of DNA replication. Although the DNA replication timing profiles are disrupted in HU-challenged cds1? cells, ~85% of potential origins overlapped with those found in wild type cells, significantly, most of which represents inefficiently fired origins in wild type cells. Together, our result indicates that replication checkpoint plays a role in monitoring efficient origins and thus maintaining global DNA replication patterns in HU-challenged cells. Keywords: WT or Cds1 HU synchronized cells released in HU free media and harvested at different time points vs WT or Cds1 synchronized with HU for 3 hrs. We analyzed 32 arrays for WT and 38 arrays for Cds1 cells which were synchronized with HU and released in HU free media and harvested at different time points. At least two biological repeats were done for each time points.

2007-08-14 | E-GEOD-6977 | biostudies-arrayexpress

Altered genes expression profile of Rep2 mutants in Fission Yeast upon HU treatment.

Project description:Expression profiles indicate that C-terminal of rep2 is essential for its transactivation activity. Keywords: rep2 mutants cells treated with 8 mM HU for different time points vs wildtype untreated cells

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Genome-wide Profiling of DNA Replication Timing in pombe.

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HU treatment of Sulfolobus solfataricus P2

Project description:Hydroxyurea (HU) is toxic to Sulfolobus cells. To address the basis of the HU toxicity, we performed transcriptome analyses on untreated cells and cells following exposure to 5 mM HU for 4 hours.

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Multiple bypass pathways of Hsk1 kinase essential for initiation of DNA replication in fission yeast

Project description:Cdc7/Hsk1 is a conserved kinase required for initiation of DNA replication that potentially regulates timing and locations of replication origin firing. Here, we show that viability of fission yeast hsk1∆ cells can be restored by loss of mrc1, which is required for maintenance of replication fork integrity, by cds1∆, or by a checkpoint-deficient mutant of mrc1. In these mutants, normally inactive origins are activated in the presence of HU and binding of Cdc45 to MCM is stimulated. mrc1∆ bypasses hsk1∆ more efficiently because of its checkpoint-independent inhibitory functions. Unexpectedly, hsk1∆ is viable at 37°C. More DNA is synthesized, and some dormant origins fire in the presence of HU at 37°C. On the other hand, hsk1∆ bypass strains grow poorly at 25°C compared to at higher temperatures. Our results show that Hsk1 functions for DNA replication can be bypassed by different genetic backgrounds as well as under varied physiological conditions, providing additional evidence for plasticity of the replication program in eukaryotes. BrdU incorporation profiles at early S-phase in mrc1∆, cds1∆ and hsk1-89 mutants.

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Altered genes expression profile of Rep2 mutants in Fission Yeast upon HU treatment.

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ChIP-on-chip Rad52 in unperturbed S phase (no HU treatment)

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2019-12-20 | E-MTAB-8605 | biostudies-arrayexpress

Project description:Cell Cycle-specific Expressions Are Modulated by Replication Checkpoint Kinases Rad3 and Cds1 in Fission Yeast

| PRJNA94775 | ENA