Identification of a second DNA binding site in the human Rad52 protein

Wataru Kagawa, Ako Kagawa, Kengo Saito, Shukuko Ikawa, Takehiko Shibata, Hitoshi Kurumizaka, Shigeyuki Yokoyama

    Research output: Contribution to journalArticle

    40 Citations (Scopus)

    Abstract

    Rad52 plays essential roles in homology-dependent double-strand break repair. Various studies have established the functions of Rad52 in Rad51-dependent and Rad51-independent repair processes. However, the precise molecular mechanisms of Rad52 in these processes remain unknown. In the present study we have identified a novel DNA binding site within Rad52 by a structure-based alanine scan mutagenesis. This site is closely aligned with the putative single-stranded DNA binding site determined previously. Mutations in this site impaired the ability of the Rad52-single-stranded DNA complex to form a ternary complex with double-stranded DNA and subsequently catalyze the formation of D-loops. We found that Rad52 introduces positive supercoils into double-stranded DNA and that the second DNA binding site is essential for this activity. Our findings suggest that Rad52 aligns two recombining DNA molecules within the first and second DNA binding sites to stimulate the homology search and strand invasion processes.

    Original languageEnglish
    Pages (from-to)24264-24273
    Number of pages10
    JournalJournal of Biological Chemistry
    Volume283
    Issue number35
    DOIs
    Publication statusPublished - 2008 Aug 29

    Fingerprint

    Rad52 DNA Repair and Recombination Protein
    Binding Sites
    DNA
    Single-Stranded DNA
    Repair
    Mutagenesis
    Alanine
    Mutation
    Molecules

    ASJC Scopus subject areas

    • Biochemistry
    • Cell Biology
    • Molecular Biology

    Cite this

    Kagawa, W., Kagawa, A., Saito, K., Ikawa, S., Shibata, T., Kurumizaka, H., & Yokoyama, S. (2008). Identification of a second DNA binding site in the human Rad52 protein. Journal of Biological Chemistry, 283(35), 24264-24273. https://doi.org/10.1074/jbc.M802204200

    Identification of a second DNA binding site in the human Rad52 protein. / Kagawa, Wataru; Kagawa, Ako; Saito, Kengo; Ikawa, Shukuko; Shibata, Takehiko; Kurumizaka, Hitoshi; Yokoyama, Shigeyuki.

    In: Journal of Biological Chemistry, Vol. 283, No. 35, 29.08.2008, p. 24264-24273.

    Research output: Contribution to journalArticle

    Kagawa, W, Kagawa, A, Saito, K, Ikawa, S, Shibata, T, Kurumizaka, H & Yokoyama, S 2008, 'Identification of a second DNA binding site in the human Rad52 protein', Journal of Biological Chemistry, vol. 283, no. 35, pp. 24264-24273. https://doi.org/10.1074/jbc.M802204200
    Kagawa W, Kagawa A, Saito K, Ikawa S, Shibata T, Kurumizaka H et al. Identification of a second DNA binding site in the human Rad52 protein. Journal of Biological Chemistry. 2008 Aug 29;283(35):24264-24273. https://doi.org/10.1074/jbc.M802204200
    Kagawa, Wataru ; Kagawa, Ako ; Saito, Kengo ; Ikawa, Shukuko ; Shibata, Takehiko ; Kurumizaka, Hitoshi ; Yokoyama, Shigeyuki. / Identification of a second DNA binding site in the human Rad52 protein. In: Journal of Biological Chemistry. 2008 ; Vol. 283, No. 35. pp. 24264-24273.
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