BRCA1 is required for maintenance of phospho-Chk1 and G2/M arrest during DNA cross-link repair in DT40 cells

Margarethe Draga, Elizabeth B. Madgett, Cassandra J. Vandenberg, David du Plessis, Aisling Kaufmann, Petra Werler, Prasun Chakraborty, Noel F. Lowndes, Kevin Hiom (Lead / Corresponding author)

    Research output: Contribution to journalArticle

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    Abstract

    The Fanconi anemia DNA repair pathway is pivotal for the efficient repair of DNA interstrand cross-links. Here, we show that FA-defective (Fancc(-)) DT40 cells arrest in G2 phase following cross-link damage and trigger apoptosis. Strikingly, cell death was reduced in Fancc(-) cells by additional deletion of the BRCA1 tumor suppressor, resulting in elevated clonogenic survival. Increased resistance to cross-link damage was not due to loss of toxic BRCA1-mediated homologous recombination but rather through the loss of a G2 checkpoint. This proapoptotic role also required the BRCA1-A complex member ABRAXAS (FAM175A). Finally, we show that BRCA1 promotes G2 arrest and cell death by prolonging phosphorylation of Chk1 on serine 345 after DNA damage to sustain arrest. Our data imply that DNA-induced cross-link death in cells defective in the FA pathway is dependent on the ability of BRCA1 to prolong cell cycle arrest in G2 phase.

    Original languageEnglish
    Pages (from-to)3829-3840
    Number of pages12
    JournalMolecular and Cellular Biology
    Volume35
    Issue number22
    Early online date31 Aug 2015
    DOIs
    Publication statusPublished - Nov 2015

    Fingerprint

    Cell Death
    G2 Phase
    Maintenance
    DNA Repair
    DNA
    Fanconi Anemia
    Poisons
    Homologous Recombination
    Cell Cycle Checkpoints
    Serine
    DNA Damage
    Phosphorylation
    Apoptosis
    Neoplasms

    Keywords

    • Animals
    • Apoptosis
    • Avian proteins
    • BRCA1 protein
    • Carrier proteins
    • Cell line
    • Chickens
    • DNA repair
    • Fanconi anemia
    • Fanconi anemia complementation group C protein
    • G2 phase cell cycle checkpoints
    • Gene deletion
    • Phosphorylation
    • Protein kinases

    Cite this

    Draga, Margarethe ; Madgett, Elizabeth B. ; Vandenberg, Cassandra J. ; du Plessis, David ; Kaufmann, Aisling ; Werler, Petra ; Chakraborty, Prasun ; Lowndes, Noel F. ; Hiom, Kevin. / BRCA1 is required for maintenance of phospho-Chk1 and G2/M arrest during DNA cross-link repair in DT40 cells. In: Molecular and Cellular Biology. 2015 ; Vol. 35, No. 22. pp. 3829-3840.
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    abstract = "The Fanconi anemia DNA repair pathway is pivotal for the efficient repair of DNA interstrand cross-links. Here, we show that FA-defective (Fancc(-)) DT40 cells arrest in G2 phase following cross-link damage and trigger apoptosis. Strikingly, cell death was reduced in Fancc(-) cells by additional deletion of the BRCA1 tumor suppressor, resulting in elevated clonogenic survival. Increased resistance to cross-link damage was not due to loss of toxic BRCA1-mediated homologous recombination but rather through the loss of a G2 checkpoint. This proapoptotic role also required the BRCA1-A complex member ABRAXAS (FAM175A). Finally, we show that BRCA1 promotes G2 arrest and cell death by prolonging phosphorylation of Chk1 on serine 345 after DNA damage to sustain arrest. Our data imply that DNA-induced cross-link death in cells defective in the FA pathway is dependent on the ability of BRCA1 to prolong cell cycle arrest in G2 phase.",
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    author = "Margarethe Draga and Madgett, {Elizabeth B.} and Vandenberg, {Cassandra J.} and {du Plessis}, David and Aisling Kaufmann and Petra Werler and Prasun Chakraborty and Lowndes, {Noel F.} and Kevin Hiom",
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    language = "English",
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    Draga, M, Madgett, EB, Vandenberg, CJ, du Plessis, D, Kaufmann, A, Werler, P, Chakraborty, P, Lowndes, NF & Hiom, K 2015, 'BRCA1 is required for maintenance of phospho-Chk1 and G2/M arrest during DNA cross-link repair in DT40 cells', Molecular and Cellular Biology, vol. 35, no. 22, pp. 3829-3840. https://doi.org/10.1128/MCB.01497-14

    BRCA1 is required for maintenance of phospho-Chk1 and G2/M arrest during DNA cross-link repair in DT40 cells. / Draga, Margarethe; Madgett, Elizabeth B.; Vandenberg, Cassandra J.; du Plessis, David; Kaufmann, Aisling; Werler, Petra; Chakraborty, Prasun; Lowndes, Noel F.; Hiom, Kevin (Lead / Corresponding author).

    In: Molecular and Cellular Biology, Vol. 35, No. 22, 11.2015, p. 3829-3840.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - BRCA1 is required for maintenance of phospho-Chk1 and G2/M arrest during DNA cross-link repair in DT40 cells

    AU - Draga, Margarethe

    AU - Madgett, Elizabeth B.

    AU - Vandenberg, Cassandra J.

    AU - du Plessis, David

    AU - Kaufmann, Aisling

    AU - Werler, Petra

    AU - Chakraborty, Prasun

    AU - Lowndes, Noel F.

    AU - Hiom, Kevin

    N1 - Copyright © 2015 Draga et al.

    PY - 2015/11

    Y1 - 2015/11

    N2 - The Fanconi anemia DNA repair pathway is pivotal for the efficient repair of DNA interstrand cross-links. Here, we show that FA-defective (Fancc(-)) DT40 cells arrest in G2 phase following cross-link damage and trigger apoptosis. Strikingly, cell death was reduced in Fancc(-) cells by additional deletion of the BRCA1 tumor suppressor, resulting in elevated clonogenic survival. Increased resistance to cross-link damage was not due to loss of toxic BRCA1-mediated homologous recombination but rather through the loss of a G2 checkpoint. This proapoptotic role also required the BRCA1-A complex member ABRAXAS (FAM175A). Finally, we show that BRCA1 promotes G2 arrest and cell death by prolonging phosphorylation of Chk1 on serine 345 after DNA damage to sustain arrest. Our data imply that DNA-induced cross-link death in cells defective in the FA pathway is dependent on the ability of BRCA1 to prolong cell cycle arrest in G2 phase.

    AB - The Fanconi anemia DNA repair pathway is pivotal for the efficient repair of DNA interstrand cross-links. Here, we show that FA-defective (Fancc(-)) DT40 cells arrest in G2 phase following cross-link damage and trigger apoptosis. Strikingly, cell death was reduced in Fancc(-) cells by additional deletion of the BRCA1 tumor suppressor, resulting in elevated clonogenic survival. Increased resistance to cross-link damage was not due to loss of toxic BRCA1-mediated homologous recombination but rather through the loss of a G2 checkpoint. This proapoptotic role also required the BRCA1-A complex member ABRAXAS (FAM175A). Finally, we show that BRCA1 promotes G2 arrest and cell death by prolonging phosphorylation of Chk1 on serine 345 after DNA damage to sustain arrest. Our data imply that DNA-induced cross-link death in cells defective in the FA pathway is dependent on the ability of BRCA1 to prolong cell cycle arrest in G2 phase.

    KW - Animals

    KW - Apoptosis

    KW - Avian proteins

    KW - BRCA1 protein

    KW - Carrier proteins

    KW - Cell line

    KW - Chickens

    KW - DNA repair

    KW - Fanconi anemia

    KW - Fanconi anemia complementation group C protein

    KW - G2 phase cell cycle checkpoints

    KW - Gene deletion

    KW - Phosphorylation

    KW - Protein kinases

    U2 - 10.1128/MCB.01497-14

    DO - 10.1128/MCB.01497-14

    M3 - Article

    VL - 35

    SP - 3829

    EP - 3840

    JO - Molecular and Cellular Biology

    JF - Molecular and Cellular Biology

    SN - 0270-7306

    IS - 22

    ER -