BRCA1-independent ubiquitination of FANCD2

Cassandra J. Vandenberg, Fanni Gergely, Chong Yi Ong, Paul Pace, Donna L. Mallery, Kevin Hiom (Lead / Corresponding author), Ketan J. Patel (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    115 Citations (Scopus)

    Abstract

    Monoubiquitination of the FANCD2 protein is a key step in the Fanconi anemia (FA) tumor suppressor pathway, coinciding with this molecule's accumulation at sites of genome damage. Strong circumstantial evidence points to a requirement for the BRCA1 gene product in this step. Here, we show that the purified BRCA1/BARD1 complex, together with E1 and UbcH5a, is sufficient to reconstitute the monoubiquitination of FANCD2 in vitro. Although siRNA-mediated knockdown of BRCA1 in human cells results in defective targeting of FANCD2 to sites of DNA damage, it does not lead to a defect in FANCD2 ubiquitination. Furthermore, ablation of the RING finger domains of either BRCA1 or BARD1 in the chicken B cell line DT40 also leaves FANCD2 modification intact. Consequently, while BRCA1 affects the accumulation of FANCD2 at sites of DNA damage, BRCA1/BARD1 E3 ligase activity is not essential for the monoubiquitination of FANCD2.

    Original languageEnglish
    Pages (from-to)247-254
    Number of pages8
    JournalMolecular Cell
    Volume12
    Issue number1
    DOIs
    Publication statusPublished - Jul 2003

    Fingerprint

    Ubiquitination
    Fanconi Anemia Complementation Group D2 Protein
    DNA Damage
    RING Finger Domains
    BRCA1 Protein
    Fanconi Anemia
    Ubiquitin-Protein Ligases
    Small Interfering RNA
    Chickens
    B-Lymphocytes
    Genome
    Cell Line
    Neoplasms
    In Vitro Techniques

    Keywords

    • Animals
    • BRCA1 protein
    • Carrier proteins
    • Cell-free system
    • DNA damage
    • Fanconi anemia
    • Fanconi anemia complementation group D2 protein
    • Gene expression regulation, Neoplastic
    • HeLa cells
    • Humans
    • Iron-binding proteins
    • Ligases
    • Mutation
    • Nuclear proteins
    • Protein structure, Tertiary
    • RNA, Small interfering
    • Tumor suppressor proteins
    • Ubiquitin
    • Ubiquitin-conjugating Enzymes
    • Ubiquitin-protein ligases

    Cite this

    Vandenberg, C. J., Gergely, F., Ong, C. Y., Pace, P., Mallery, D. L., Hiom, K., & Patel, K. J. (2003). BRCA1-independent ubiquitination of FANCD2. Molecular Cell, 12(1), 247-254. https://doi.org/10.1016/S1097-2765(03)00281-8
    Vandenberg, Cassandra J. ; Gergely, Fanni ; Ong, Chong Yi ; Pace, Paul ; Mallery, Donna L. ; Hiom, Kevin ; Patel, Ketan J. / BRCA1-independent ubiquitination of FANCD2. In: Molecular Cell. 2003 ; Vol. 12, No. 1. pp. 247-254.
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    abstract = "Monoubiquitination of the FANCD2 protein is a key step in the Fanconi anemia (FA) tumor suppressor pathway, coinciding with this molecule's accumulation at sites of genome damage. Strong circumstantial evidence points to a requirement for the BRCA1 gene product in this step. Here, we show that the purified BRCA1/BARD1 complex, together with E1 and UbcH5a, is sufficient to reconstitute the monoubiquitination of FANCD2 in vitro. Although siRNA-mediated knockdown of BRCA1 in human cells results in defective targeting of FANCD2 to sites of DNA damage, it does not lead to a defect in FANCD2 ubiquitination. Furthermore, ablation of the RING finger domains of either BRCA1 or BARD1 in the chicken B cell line DT40 also leaves FANCD2 modification intact. Consequently, while BRCA1 affects the accumulation of FANCD2 at sites of DNA damage, BRCA1/BARD1 E3 ligase activity is not essential for the monoubiquitination of FANCD2.",
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    Vandenberg, CJ, Gergely, F, Ong, CY, Pace, P, Mallery, DL, Hiom, K & Patel, KJ 2003, 'BRCA1-independent ubiquitination of FANCD2', Molecular Cell, vol. 12, no. 1, pp. 247-254. https://doi.org/10.1016/S1097-2765(03)00281-8

    BRCA1-independent ubiquitination of FANCD2. / Vandenberg, Cassandra J.; Gergely, Fanni; Ong, Chong Yi; Pace, Paul; Mallery, Donna L.; Hiom, Kevin (Lead / Corresponding author); Patel, Ketan J. (Lead / Corresponding author).

    In: Molecular Cell, Vol. 12, No. 1, 07.2003, p. 247-254.

    Research output: Contribution to journalArticle

    TY - JOUR

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    AU - Vandenberg, Cassandra J.

    AU - Gergely, Fanni

    AU - Ong, Chong Yi

    AU - Pace, Paul

    AU - Mallery, Donna L.

    AU - Hiom, Kevin

    AU - Patel, Ketan J.

    PY - 2003/7

    Y1 - 2003/7

    N2 - Monoubiquitination of the FANCD2 protein is a key step in the Fanconi anemia (FA) tumor suppressor pathway, coinciding with this molecule's accumulation at sites of genome damage. Strong circumstantial evidence points to a requirement for the BRCA1 gene product in this step. Here, we show that the purified BRCA1/BARD1 complex, together with E1 and UbcH5a, is sufficient to reconstitute the monoubiquitination of FANCD2 in vitro. Although siRNA-mediated knockdown of BRCA1 in human cells results in defective targeting of FANCD2 to sites of DNA damage, it does not lead to a defect in FANCD2 ubiquitination. Furthermore, ablation of the RING finger domains of either BRCA1 or BARD1 in the chicken B cell line DT40 also leaves FANCD2 modification intact. Consequently, while BRCA1 affects the accumulation of FANCD2 at sites of DNA damage, BRCA1/BARD1 E3 ligase activity is not essential for the monoubiquitination of FANCD2.

    AB - Monoubiquitination of the FANCD2 protein is a key step in the Fanconi anemia (FA) tumor suppressor pathway, coinciding with this molecule's accumulation at sites of genome damage. Strong circumstantial evidence points to a requirement for the BRCA1 gene product in this step. Here, we show that the purified BRCA1/BARD1 complex, together with E1 and UbcH5a, is sufficient to reconstitute the monoubiquitination of FANCD2 in vitro. Although siRNA-mediated knockdown of BRCA1 in human cells results in defective targeting of FANCD2 to sites of DNA damage, it does not lead to a defect in FANCD2 ubiquitination. Furthermore, ablation of the RING finger domains of either BRCA1 or BARD1 in the chicken B cell line DT40 also leaves FANCD2 modification intact. Consequently, while BRCA1 affects the accumulation of FANCD2 at sites of DNA damage, BRCA1/BARD1 E3 ligase activity is not essential for the monoubiquitination of FANCD2.

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    KW - Carrier proteins

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    KW - Fanconi anemia complementation group D2 protein

    KW - Gene expression regulation, Neoplastic

    KW - HeLa cells

    KW - Humans

    KW - Iron-binding proteins

    KW - Ligases

    KW - Mutation

    KW - Nuclear proteins

    KW - Protein structure, Tertiary

    KW - RNA, Small interfering

    KW - Tumor suppressor proteins

    KW - Ubiquitin

    KW - Ubiquitin-conjugating Enzymes

    KW - Ubiquitin-protein ligases

    U2 - 10.1016/S1097-2765(03)00281-8

    DO - 10.1016/S1097-2765(03)00281-8

    M3 - Article

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    JO - Molecular Cell

    JF - Molecular Cell

    SN - 1097-2765

    IS - 1

    ER -

    Vandenberg CJ, Gergely F, Ong CY, Pace P, Mallery DL, Hiom K et al. BRCA1-independent ubiquitination of FANCD2. Molecular Cell. 2003 Jul;12(1):247-254. https://doi.org/10.1016/S1097-2765(03)00281-8