Region and amino acid residues required for Rad51C binding in the human Xrcc3 protein

Hitoshi Kurumizaka, Rima Enomoto, Maki Nakada, Keiko Eda, Shigeyuki Yokoyama, Takehiko Shibata

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    15 Citations (Scopus)

    Abstract

    The Xrcc3 protein, which is required for the homologous recombinational repair of damaged DNA, forms a complex with the Rad51C protein in human cells. Mutations in either the Xrcc3 or Rad51C gene cause extreme sensitivity to DNA-damaging agents and generate the genomic instability frequently found in tumors. In the present study, we found that the Xrcc3 segment containing amino acid residues 63-346, Xrcc363-346, is the Rad51C-binding region. Biochemical analyses revealed that Xrcc363-346 forms a complex with Rad51C, and the Xrcc363-346-Rad51C complex possesses ssDNA and dsDNA binding abilities comparable to those of the full-length Xrcc3-Rad51C complex. Based on the structure of RecA, which is thought to be the ancestor of Xrcc3, six Xrcc3 point mutants were designed. Two-hybrid and biochemical analyses of the Xrcc3 point mutants revealed that Tyr139 and Phe249 are essential amino acid residues for Rad51C binding. Superposition of the Xrcc3 Tyr139 and Phe249 residues on the RecA structure suggested that Tyr139 may function to ensure proper folding and Phe249 may be important to constitute the Rad51C-binding interface in Xrcc3.

    Original languageEnglish
    Pages (from-to)4041-4050
    Number of pages10
    JournalNucleic Acids Research
    Volume31
    Issue number14
    DOIs
    Publication statusPublished - 2003 Jul 15

    ASJC Scopus subject areas

    • Genetics

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  • Cite this

    Kurumizaka, H., Enomoto, R., Nakada, M., Eda, K., Yokoyama, S., & Shibata, T. (2003). Region and amino acid residues required for Rad51C binding in the human Xrcc3 protein. Nucleic Acids Research, 31(14), 4041-4050. https://doi.org/10.1093/nar/gkg442