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)

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    116 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

    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

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  • 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