Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Chromatin immunopreciptiation of human lymphoblastoid untreated and treated with aphidicolin (APH), trichostatin A (TSA), or APH plus TSA cells to map the origins of DNA replication at common fragile sites

ABSTRACT: Map the origins of DNA replication in human cells at 7 common fragile sites and their flanking non-fragile sequences as well as a 200kb containing the rare fragile site FRAXA, and a 1,075kb non-fragile region on chr22 . The origins were mapped in untreated cells, as well as, in cells treated with aphidicolin (APH), trichostatin A (TSA), or APH plus TSA. The origin mapping experiment is based on the fact that during S phase, short newly replicated DNA fragments (300bp-1kb) are generated only at the origins of replication. By combining the nascent strand assay with a tiled microarray platform, we have previoulsy developed a rapid, non-PCR based, high-throughput approach to map active origins in asynchronous human cells (Lucas et al., 2007, AC 17668008).

ORGANISM(S): Homo sapiens

SUBMITTER: Isabelle Lucas

PROVIDER: E-TABM-348 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

ACCESS DATA

Publications

Impaired replication dynamics at the FRA3B common fragile site.

Palakodeti Aparna A Lucas Isabelle I Jiang Yanwen Y Young David J DJ Fernald Anthony A AA Karrison Theodore T Le Beau Michelle M MM

Human molecular genetics 20100101 1

Chromosomal common fragile sites (CFSs) are genetically unstable regions of the genome that are induced by conditions that impair DNA replication. In this report, we show that treatment with the DNA polymerase inhibitor, aphidicolin (APH), slows the replication rate throughout S phase. To investigate the unusual sensitivity of CFSs to APH-induced replication stress, we examined replication dynamics within a 50 kb region of the most frequently expressed CFS, FRA3B. We mapped four origins of repli ...[more]

PMID: 19815620

Similar Datasets

Project description:Chromosomal DNA replication involves the coordinated activity of hundreds to thousands of replication origins. Individual replication origins are subject to epigenetic regulation of their activity during S-phase, resulting in differential efficiencies and timings of replication initiation during S-phase. This regulation is thought to involve chromatin structure and organization into timing domains with differential ability to recruit limiting replication factors. Rif1 has recently been identified as a genome-wide regulator of replication timing in fission yeast and in mammalian cells. However, previous studies in budding yeast have suggested that Rif1’s role in controlling replication timing may be limited to subtelomeric domains and derives from its established role in telomere length regulation. We have analyzed replication timing by analyzing BrdU incorporation genome-wide, and report that Rif1 regulates the timing of late/dormant replication origins throughout the S. cerevisiae genome. Analysis of pfa4∆ cells, which are defective in palmitoylation and membrane association of Rif1, suggests that replication timing regulation by Rif1 is independent of its role in localizing telomeres to the nuclear periphery. Intra-S checkpoint signaling is intact in rif1∆ cells, and checkpoint-defective mec1∆ cells do not comparably deregulate replication timing, together indicating that Rif1 regulates replication timing through a mechanism independent of this checkpoint. Our results indicate that the Rif1 mechanism regulates origin timing irrespective of proximity to a chromosome end, and suggest instead that telomere sequences merely provide abundant binding sites for proteins that recruit Rif1. Still, the abundance of Rif1 binding in telomeric domains may facilitate Rif1-mediated repression of non-telomeric origins that are more distal from centromeres. 30 total samples: (6 samples - BrdU- HU arrest 45min with 2 replicates, strains: WT, rif1 delta, pfa4 delta) (12 samples -S-phase BrdU time course with 2 replicates at 25 and 35 min, strains: WT, rif1 delta, mec1_100) (12 samples - S-phase BrdU time course with 2 replicates at 25 and 35 min, strains: sml1 delta, sml1 delta rif1 delta, sml1 delta mec1 delta)

Dataset Information

Chromatin immunopreciptiation of human lymphoblastoid untreated and treated with aphidicolin (APH), trichostatin A (TSA), or APH plus TSA cells to map the origins of DNA replication at common fragile sites

Publications

Impaired replication dynamics at the FRA3B common fragile site.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets