Single-stranded DNA catenation mediated by human EVL and a type i topoisomerase

Motoki Takaku, Daisuke Takahashi, Shinichi MacHida, Hiroyuki Ueno, Noriko Hosoya, Shukuko Ikawa, Kiyoshi Miyagawa, Takehiko Shibata, Hitoshi Kurumizaka

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    Abstract

    The human Ena/Vasp-like (EVL) protein is considered to be a bifunctional protein, involved in both actin remodeling and homologous recombination. In the present study, we found that human EVL forms heat-stable multimers of circular single-stranded DNA (ssDNA) molecules in the presence of a type I topoisomerase in vitro. An electron microscopic analysis revealed that the heat-stable ssDNA multimers formed by EVL and topoisomerase were ssDNA catemers. The ssDNA catenation did not occur when either EVL or topoisomerase was omitted from the reaction mixture. A deletion analysis revealed that the ssDNA catenation completely depended on the annealing activity of EVL. Human EVL was captured from a human cell extract by TOPO IIIα-conjugated beads, and the interaction between EVL and TOPO IIIα was confirmed by a surface plasmon resonance analysis. Purified TOPO IIIα catalyzed the ssDNA catenation with EVL as efficiently as the Escherichia coli topoisomerase I. Since the ssDNA cutting and rejoining reactions, which are the sub-steps of ssDNA catenation, may be an essential process in homologous recombination, EVL and TOPO IIIα may function in the processing of DNA intermediates formed during homologous recombination.

    Original languageEnglish
    Pages (from-to)7579-7586
    Number of pages8
    JournalNucleic Acids Research
    Volume38
    Issue number21
    DOIs
    Publication statusPublished - 2010 Nov

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    ASJC Scopus subject areas

    • Genetics

    Cite this

    Takaku, M., Takahashi, D., MacHida, S., Ueno, H., Hosoya, N., Ikawa, S., Miyagawa, K., Shibata, T., & Kurumizaka, H. (2010). Single-stranded DNA catenation mediated by human EVL and a type i topoisomerase. Nucleic Acids Research, 38(21), 7579-7586. https://doi.org/10.1093/nar/gkq630