BRCA2 and RAD51 promote double-strand break formation and cell death in response to Gemcitabine

Rebecca M Jones; Panagiotis Kotsantis; Grant Stewart; Petra Groth; Eva Petermann

doi:10.1158/1535-7163.MCT-13-0862

BRCA2 and RAD51 promote double-strand break formation and cell death in response to Gemcitabine

Rebecca M Jones
, Panagiotis Kotsantis
, Grant Stewart
, Petra Groth
, Eva Petermann

University of Birmingham
Karolinska Institutet

Research output: Contribution to journal › Article › peer-review

39 Citations (Scopus)

Abstract

Replication inhibitors cause replication fork stalling and double-strand breaks (DSBs) that result from processing of stalled forks. During recovery from replication blocks, the homologous recombination (HR) factor RAD51 mediates fork restart and DSB repair. HR defects therefore sensitise cells to replication inhibitors, with clear implications for cancer therapy. Gemcitabine is a potent replication inhibitor used to treat cancers with mutations in HR genes such as BRCA2. Here we investigate why, paradoxically, mutations in HR genes protect cells from killing by Gemcitabine. Using DNA replication and -damage assays in mammalian cells, we show that even short Gemcitabine treatments cause persistent replication inhibition. BRCA2 and RAD51 are recruited to chromatin early after removal of the drug, actively inhibit replication fork progression and promote the formation of MUS81- and XPF-dependent DSBs that remain unrepaired. Our data suggest that HR intermediates formed at Gemcitabine-stalled forks are converted into DSBs and thus contribute to Gemcitabine-induced cell death, which could have implications for the treatment response of HR-deficient tumours.

Original language	English
Pages (from-to)	2412-2421
Number of pages	10
Journal	Molecular Cancer Therapeutics
Volume	13
Issue number	10
DOIs	https://doi.org/10.1158/1535-7163.MCT-13-0862
Publication status	Published - 2 Oct 2014

Funding

This work was supported by the Medical Research Council (MR/J007595/1; to E. Petermann), the Association for International Cancer Research (13-1048; to E. Petermann), Wellcome Trust (ISSFPP12; to E. Petermann), Cancer Research UK (C17183/A13030; to G.S. Stewart). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

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SDG 3 Good Health and Well-being

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title = "BRCA2 and RAD51 promote double-strand break formation and cell death in response to Gemcitabine",

abstract = "Replication inhibitors cause replication fork stalling and double-strand breaks (DSBs) that result from processing of stalled forks. During recovery from replication blocks, the homologous recombination (HR) factor RAD51 mediates fork restart and DSB repair. HR defects therefore sensitise cells to replication inhibitors, with clear implications for cancer therapy. Gemcitabine is a potent replication inhibitor used to treat cancers with mutations in HR genes such as BRCA2. Here we investigate why, paradoxically, mutations in HR genes protect cells from killing by Gemcitabine. Using DNA replication and -damage assays in mammalian cells, we show that even short Gemcitabine treatments cause persistent replication inhibition. BRCA2 and RAD51 are recruited to chromatin early after removal of the drug, actively inhibit replication fork progression and promote the formation of MUS81- and XPF-dependent DSBs that remain unrepaired. Our data suggest that HR intermediates formed at Gemcitabine-stalled forks are converted into DSBs and thus contribute to Gemcitabine-induced cell death, which could have implications for the treatment response of HR-deficient tumours.",

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T1 - BRCA2 and RAD51 promote double-strand break formation and cell death in response to Gemcitabine

AU - Jones, Rebecca M

AU - Kotsantis, Panagiotis

AU - Stewart, Grant

AU - Groth, Petra

AU - Petermann, Eva

PY - 2014/10/2

Y1 - 2014/10/2

N2 - Replication inhibitors cause replication fork stalling and double-strand breaks (DSBs) that result from processing of stalled forks. During recovery from replication blocks, the homologous recombination (HR) factor RAD51 mediates fork restart and DSB repair. HR defects therefore sensitise cells to replication inhibitors, with clear implications for cancer therapy. Gemcitabine is a potent replication inhibitor used to treat cancers with mutations in HR genes such as BRCA2. Here we investigate why, paradoxically, mutations in HR genes protect cells from killing by Gemcitabine. Using DNA replication and -damage assays in mammalian cells, we show that even short Gemcitabine treatments cause persistent replication inhibition. BRCA2 and RAD51 are recruited to chromatin early after removal of the drug, actively inhibit replication fork progression and promote the formation of MUS81- and XPF-dependent DSBs that remain unrepaired. Our data suggest that HR intermediates formed at Gemcitabine-stalled forks are converted into DSBs and thus contribute to Gemcitabine-induced cell death, which could have implications for the treatment response of HR-deficient tumours.

AB - Replication inhibitors cause replication fork stalling and double-strand breaks (DSBs) that result from processing of stalled forks. During recovery from replication blocks, the homologous recombination (HR) factor RAD51 mediates fork restart and DSB repair. HR defects therefore sensitise cells to replication inhibitors, with clear implications for cancer therapy. Gemcitabine is a potent replication inhibitor used to treat cancers with mutations in HR genes such as BRCA2. Here we investigate why, paradoxically, mutations in HR genes protect cells from killing by Gemcitabine. Using DNA replication and -damage assays in mammalian cells, we show that even short Gemcitabine treatments cause persistent replication inhibition. BRCA2 and RAD51 are recruited to chromatin early after removal of the drug, actively inhibit replication fork progression and promote the formation of MUS81- and XPF-dependent DSBs that remain unrepaired. Our data suggest that HR intermediates formed at Gemcitabine-stalled forks are converted into DSBs and thus contribute to Gemcitabine-induced cell death, which could have implications for the treatment response of HR-deficient tumours.

UR - https://aacrjournals.org/mct/article/13/10/2412/128821/BRCA2-and-RAD51-Promote-Double-Strand-Break

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DO - 10.1158/1535-7163.MCT-13-0862

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JO - Molecular Cancer Therapeutics

JF - Molecular Cancer Therapeutics

IS - 10

ER -

BRCA2 and RAD51 promote double-strand break formation and cell death in response to Gemcitabine

Abstract

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UN SDGs

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