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Genome-wide binding of hSTN1 in response to hydroxyurea-induced replication stress

ABSTRACT: We find that hSTN1 is preferentially enriched at GC-rich repetitive sequences genome-wide after hydroxyurea treatment.

ORGANISM(S): Homo sapiens

PROVIDER: GSE82123 | GEO | 2016/08/06

SECONDARY ACCESSION(S): PRJNA324101

REPOSITORIES: GEO

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Genome-wide binding of hSTN1 in response to hydroxyurea-induced replication stress

Project description:We find that hSTN1 is preferentially enriched at GC-rich repetitive sequences genome-wide after hydroxyurea treatment. HeLa cells stably expressing myc-tagged hSTN1 were treated with 2 mM hydroxyurea, crosslinked, and ChIP-seq was performed.

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Project description:To investigate the gene expression changes in gestational day 9 embryos exposed to hydroxyurea in utero. Agilent microarray technology was used to identify genes that responded to hydroxyurea exposure. Understanding the stress response of the embryo will help us advance education of birth defect prevention. Gestational day 9 embryos were exposed to 400 mg/kg hydroxyurea, a dose that induces a moderate number of malformations, including fetal resorptions, growth retardation, external and skeletal defects. We identified several genes involved in responding to oxidative stress, including NRF-2, PRDX1, and TXNIP. Other genes also significantly changed were involved in cell cycle arrest and apoptosis. The dysregulation in gene expression during organogenesis contributes to the developmental toxicity of hydroxyurea. Gestational day 9 mouse embryos were treated with saline or 400 mg/kg hydroxyurea by intraperitoneal injection. Dams were euthanized after 3 hours and embryos explanted. 6 indendent experiments were performed for each sample.

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Project description:Mammalian DNA replication relies on various DNA helicases and nuclease activities to ensure accurate genetic duplication, but how different helicase and nuclease activities are properly directed remains unclear. Here, we identify the ubiquitin-specific protease, USP50, as a chromatin-associated protein required to promote ongoing replication, fork restart, telomere maintenance, cellular survival following hydroxyurea or pyridostatin treatment, and suppression of DNA breaks near GC-rich sequences. We find that USP50 supports proper WRN: FEN1 localisation at or near stalled replication forks. Nascent DNA in cells lacking USP50 shows increased association of the DNA2 nuclease and RECQL4 and RECQL5 helicases and replication defects in cells lacking USP50, or FEN1 are driven by these proteins. Consequently, suppression of DNA2 or RECQL4/5 improves USP50-depleted cell resistance to agents inducing replicative stress and restores telomere stability. These data define an unexpected regulatory protein that promotes the balance of helicase and nuclease use at ongoing and stalled replication forks..

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Project description:Multicenter Study of Hydroxyurea (MSH)

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Project description:Multicenter Study of Hydroxyurea (MSH)

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