ChIP-on-chip Rad52 in unperturbed S phase (no HU treatment)
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA418980 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:ChIP-on-chip of Rad52 at time 90' after cdc25-22 release in the following strains: cdc25-22, cdc25-22 rad3∆, cdc25-22 rif1∆ and cdc25-22 rif1∆rad3∆
2018-05-31 | GSE107098 | GEO
Project description:Checkpoints are cellular surveillance and signaling pathways that regulate responses to DNA damage and perturbations of DNA replication. Here we show that high levels of sumoylated Rad52 are present in the mec1 sml1 and rad53 sml1 checkpoint mutants exposed to DNA damaging agents such as methyl methanesulfonate (MMS) or the DNA replication inhibitor hydroxyurea (HU). The kinase-defective mutant rad53-K227A also showed high levels of Rad52 sumoylation. Elevated levels of Rad52 sumoylation occur in checkpoint mutants proceeding S phase being exposed DNA-damaging agent. Interestingly, ChIP on chip analyses revealed non-canonical chromosomal localization of Rad52 in the HU-treated rad53-K227A cells arrested in early S phase: Rad52 localization at dormant and early DNA replication origins. However, such unusual localization was not dependent on the sumoylation of Rad52. In addition, we also found that Rad52 could be highly sumoylated in the absence of Rad51. Double deletion of RAD51 and RAD53 exhibited the similar levels of Rad52 sumoylation to RAD51 single deletion. The significance and regulation mechanism of Rad52 sumoylation by checkpoint pathways will be discussed. Keywords: ChIP-chip
2009-05-26 | GSE14761 | GEO
Project description:ChIP-on-chip Rad52 in early S phase (time 60min)
| PRJNA451509 | ENA
Project description:Checkpoints are cellular surveillance and signaling pathways that regulate responses to DNA damage and perturbations of DNA replication. Here we show that high levels of sumoylated Rad52 are present in the mec1 sml1 and rad53 sml1 checkpoint mutants exposed to DNA damaging agents such as methyl methanesulfonate (MMS) or the DNA replication inhibitor hydroxyurea (HU). The kinase-defective mutant rad53-K227A also showed high levels of Rad52 sumoylation. Elevated levels of Rad52 sumoylation occur in checkpoint mutants proceeding S phase being exposed DNA-damaging agent. Interestingly, ChIP on chip analyses revealed non-canonical chromosomal localization of Rad52 in the HU-treated rad53-K227A cells arrested in early S phase: Rad52 localization at dormant and early DNA replication origins. However, such unusual localization was not dependent on the sumoylation of Rad52. In addition, we also found that Rad52 could be highly sumoylated in the absence of Rad51. Double deletion of RAD51 and RAD53 exhibited the similar levels of Rad52 sumoylation to RAD51 single deletion. The significance and regulation mechanism of Rad52 sumoylation by checkpoint pathways will be discussed. Keywords: ChIP-chip ⢠The goal of the experiment Genome-wide localization of Rad52 binding sites in Saccharomyces cerevisiae ⢠Experimental factor Distribution of Rad52 on chromosome III, IV, and V and the right arm of chromosome VI Strain: wild type, rad53 mutant, and rad53 siz2 mutant (W303 background, expressing myc tagged protein from its native promoter) Cell condition 1: G1 arrest with alpha-factor Cell condition 2: HU treatment ⢠Experimental design ChIP analyses: ChIP using anti-c-Myc antibody. ChIP-chip analyses: In all cases, hybridization data of ChIP fraction was compared with WCE (whole cell extract) fraction. Saccharomyces cerevisiae affymetrix genome tiling array (SC3456a520015F) were used. ⢠Quality control steps taken Confirmation of several loci by quantitative real time PCR. Wild type cells expressing non-tagged Rad52 were used as a negative control of DNA amplification.
2010-05-18 | E-GEOD-14761 | biostudies-arrayexpress
Project description:ChIP-seq profile of RPA in B upon HU treatment
| PRJNA478137 | ENA
Project description:Determination of the genome-wide distribution of ORC by chromatin immunoprecipitation in the Drosophila Kc167 cell line at the beginning of S phase. Kc167 cells were arrested at the G1/S transition with hydroxyurea (HU). Goal was to test whether the ORC binding distribution remained the same between G1 and S. ChIP-Chip of ORC in HU compared to input genomic DNA. Biological Replicates: 1
2014-12-05 | E-GEOD-63914 | biostudies-arrayexpress
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.
2016-10-05 | GSE87612 | GEO
Project description:Determination of the genome-wide distribution of the Mcm2-7 helicase by chromatin immunoprecipitation in the Drosophila Kc167 cell line at the beginning of S phase. Kc167 cells were arrested at the G1/S transition with hydroxyurea (HU). Goal was to evaluate changes in the genome-wide Mcm2-7 distribution throughout the cell cycle, and how this relates to the excess Mcm2-7 loaded onto chromatin in G1. ChIP-Chip of Mcm2-7 in HU compared to input genomic DNA. Biological Replicates: 2
2014-11-27 | E-GEOD-63680 | biostudies-arrayexpress
Project description:Total S phase was measured for wild-type cells undergoing meiS and mitS. Early replication origins were mapped in mitS in wild-type cells, and in meiS for wild-type, sml1 delete, rec8 delete and spo11 delete cells. Comparative genome hybridization was used to measure the kinetics and final extent of DNA replication in wild-type and mutant cells. First, S phase profiles were produced from wild-type cells undergoing pre-mitotic S phase (mitS) and pre-meiotic S phase (meiS) to measure the kinetics of DNA replication in the two S phases. For these experiments, a pool of samples collected throught out all of S phase were labeled and hybridized to a microarray together with a control G1 DNA sample. Second, early replication origins were mapped by exposing cells to levels of HU that strongly reduced DNA replication. Early replication was compared for wild-type cells in mitS to wild-type, sml1, rec8 and spo11 delete strain undergoing meiS. In these experiment, samples were collected after 4 hours exposure to HU. Total genomic DNA was isolated, labeled and hybridized to a microarray together with a control G1 DNA sample. A control G1 vs. G1 sample was prepared to measure the level of noise inherent in the CGH technique.
2012-06-18 | E-GEOD-35662 | biostudies-arrayexpress
Project description:ChIP-on-chip of Rad52 at time 60' after cdc25-22 release in the following strains: cdc25-22, cdc25-22 rad3∆
2018-05-31 | GSE113559 | GEO