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Genome-wide fork-collapse sites in MDA-MB-231 cells from ATR inhibition [human]

ABSTRACT: Genome-wide fork-collapse sites in MDA-MB-231 cells from ATR inhibition [human]

PROVIDER: PRJNA475583 | ENA |

REPOSITORIES: ENA

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Genome-wide fork-collapse sites in MDA-MB-231 cells from ATR inhibition [human]

Project description:Transient obstruction of DNA polymerase progression activates the ATR checkpoint kinase, which suppresses fork breakage, strand resection, and RPA accumulation. Herein, we use a developed DNA break-detection assay, BrITL, to identify replication-problematic loci that become processed into persistent double-strand breaks across the human genome from ATR inhibition.

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Genome-wide fork-collapse sites in MEF and MDA-MB-231 from ATR inhibition

Project description:Genome-wide fork-collapse sites in MEF and MDA-MB-231 from ATR inhibition

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Genome-wide fork-collapse sites in MEFs from ATR inhibition

Project description:Transient obstruction of DNA polymerase progression activates the ATR checkpoint kinase, which suppresses fork breakage, strand resection, and RPA accumulation. Herein, we use RPA ChIP-Seq to identify replication-problematic loci (RPLs) across the mammalian genome from ATR inhibition.

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Project description:Transient obstruction of DNA polymerase progression activates the ATR checkpoint kinase, which suppresses fork breakage, strand resection, and RPA accumulation. Herein, we use a developed DNA break-detection assay, BrITL, to identify replication-problematic loci (RPLs) that become processed into persistent double-strand breaks across the mammalian genome from ATR inhibition.

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Genome-wide fork-collapse sites in MEFs from ATR inhibition [mouse]

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Project description:This SuperSeries is composed of the SubSeries listed below.

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Checkpoint-mediated DNA polymerase e exonuclease activity curbing counteracts resection-driven fork collapse

Project description:DNA polymerase epsilon (Pole) carries out leading strand synthesis with high fidelity owing to its exonuclease activity. Pole polymerase and exonuclease activities are in balance, due to partitioning of nascent strands between catalytic sites, so that net end resection occurs when synthesis is impaired. Stalling of chromosomal DNA synthesis activates replication checkpoint kinases, required to preserve the functional integrity of replication forks. We found that Pole is phosphorylated in a Rad53CHK1-dependent manner upon fork stalling, likely to limit Pole-driven nascent strand resection that causes replication fork collapse. In stress conditions Pole phosphorylation occurs on serine 430 of the Pol2 catalytic subunit. A S430 phosphomimic limits strand partitioning and exonucleolytic processivity, while non-phosphorylatable Pol2-S430A bypasses checkpoint regulation causing stalled fork resection and collapse. We propose that checkpoint kinases switch Pole to an exonuclease-safe mode by curbing active site partitioning thus preventing nascent strand resection and stabilizing stalled replication forks.

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