The putative nuclear localization signal of the human RAD52 protein is a potential sumoylation site

Kengo Saito, Wataru Kagawa, Takehiro Suzuki, Hidekazu Suzuki, Shigeyuki Yokoyama, Hisato Saitoh, Satoshi Tashiro, Naoshi Dohmae, Hitoshi Kurumizaka

    Research output: Contribution to journalArticle

    15 Citations (Scopus)

    Abstract

    RAD52, a key factor in homologous recombination (HR), plays important roles in both RAD51-dependent and -independent HR pathways. Several studies have suggested a link between the functional regulation of RAD52 and the protein modification by a small ubiquitin-like modifier (SUMO). However, the molecular mechanism underlying the regulation of RAD52 by SUMO is unknown. To begin investigating this mechanism, we identified possible target sites for sumoylation in the human RAD52 protein by preparing a RAD52-SUMO complex using an established Escherichia coli sumoylation system. Mass spectrometry and amino acid sequencing of the enzymatically digested fragments of the purified complex revealed that the putative nuclear localization signal located near the C terminus of RAD52 was sumoylated. Biochemical studies of the RAD52-SUMO complex suggested that sumoylation at the identified site has no apparent effect on the DNA binding, D-loop formation, ssDNA annealing and RAD51-binding activities of RAD52. On the other hand, visualization of the GFP-fused RAD52 protein in the human cell that contained mutations at the identified sumoylation sites showed clear differences in the cytosolic and nuclear distributions of the protein. These results suggest the possibility of sumoylation playing an important role in the nuclear transport of RAD52.

    Original languageEnglish
    Pages (from-to)833-842
    Number of pages10
    JournalJournal of Biochemistry
    Volume147
    Issue number6
    DOIs
    Publication statusPublished - 2010 Jun

    Fingerprint

    Sumoylation
    Nuclear Localization Signals
    Ubiquitin
    Homologous Recombination
    Proteins
    Cell Nucleus Active Transport
    Escherichia coli
    Mass spectrometry
    Protein Sequence Analysis
    Nuclear Proteins
    Visualization
    Cells
    Annealing
    Mass Spectrometry
    Amino Acids
    human RAD52 protein
    DNA
    Mutation

    Keywords

    • enzymes
    • homologous recombination
    • post translational modification
    • RAD52
    • self-association
    • sumoylation

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology

    Cite this

    Saito, K., Kagawa, W., Suzuki, T., Suzuki, H., Yokoyama, S., Saitoh, H., ... Kurumizaka, H. (2010). The putative nuclear localization signal of the human RAD52 protein is a potential sumoylation site. Journal of Biochemistry, 147(6), 833-842. https://doi.org/10.1093/jb/mvq020

    The putative nuclear localization signal of the human RAD52 protein is a potential sumoylation site. / Saito, Kengo; Kagawa, Wataru; Suzuki, Takehiro; Suzuki, Hidekazu; Yokoyama, Shigeyuki; Saitoh, Hisato; Tashiro, Satoshi; Dohmae, Naoshi; Kurumizaka, Hitoshi.

    In: Journal of Biochemistry, Vol. 147, No. 6, 06.2010, p. 833-842.

    Research output: Contribution to journalArticle

    Saito, K, Kagawa, W, Suzuki, T, Suzuki, H, Yokoyama, S, Saitoh, H, Tashiro, S, Dohmae, N & Kurumizaka, H 2010, 'The putative nuclear localization signal of the human RAD52 protein is a potential sumoylation site', Journal of Biochemistry, vol. 147, no. 6, pp. 833-842. https://doi.org/10.1093/jb/mvq020
    Saito, Kengo ; Kagawa, Wataru ; Suzuki, Takehiro ; Suzuki, Hidekazu ; Yokoyama, Shigeyuki ; Saitoh, Hisato ; Tashiro, Satoshi ; Dohmae, Naoshi ; Kurumizaka, Hitoshi. / The putative nuclear localization signal of the human RAD52 protein is a potential sumoylation site. In: Journal of Biochemistry. 2010 ; Vol. 147, No. 6. pp. 833-842.
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