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 journalArticlepeer-review

    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

    Keywords

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

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology

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