Direct observation of strand passage by dna-topoisomerase and its limited processivity

Katsunori Yogo, Taisaku Ogawa, Masahito Hayashi, Yoshie Harada, Takayuki Nishizaka, Kazuhiko Kinosita

    Research output: Contribution to journalArticle

    8 Citations (Scopus)

    Abstract

    Type-II DNA topoisomerases resolve DNA entanglements such as supercoils, knots and catenanes by passing one segment of DNA duplex through a transient enzyme-bridged double-stranded break in another segment. The ATP-dependent passage reaction has previously been demonstrated at the single-molecule level, showing apparent processivity at saturating ATP. Here we directly observed the strand passage by human topoisomerase IIα, after winding a pair of fluorescently stained DNA molecules with optical tweezers for 30 turns into an X-shaped braid. On average 0.51±0.33 μm (11±6 turns) of a braid was unlinked in a burst of reactions taking 8±4 s, the unlinked length being essentially independent of the enzyme concentration between 0.25-37 pM. The time elapsed before the start of processive unlinking decreased with the enzyme concentration, being ~100 s at 3.7 pM. These results are consistent with a scenario where the enzyme binds to one DNA for a period of ~10 s, waiting for multiple diffusional encounters with the other DNA to transport it across the break ~10 times, and then dissociates from the binding site without waiting for the exhaustion of transportable DNA segments.

    Original languageEnglish
    Article numbere34920
    JournalPLoS One
    Volume7
    Issue number4
    DOIs
    Publication statusPublished - 2012 Apr 9

    Fingerprint

    Observation
    DNA
    DNA topoisomerase (ATP-hydrolysing)
    Type II DNA Topoisomerase
    Enzymes
    enzymes
    Catenanes
    Adenosine Triphosphate
    Optical Tweezers
    Optical tweezers
    Molecules
    knots
    binding sites
    Binding Sites

    ASJC Scopus subject areas

    • Agricultural and Biological Sciences(all)
    • Biochemistry, Genetics and Molecular Biology(all)
    • Medicine(all)

    Cite this

    Yogo, K., Ogawa, T., Hayashi, M., Harada, Y., Nishizaka, T., & Kinosita, K. (2012). Direct observation of strand passage by dna-topoisomerase and its limited processivity. PLoS One, 7(4), [e34920]. https://doi.org/10.1371/journal.pone.0034920

    Direct observation of strand passage by dna-topoisomerase and its limited processivity. / Yogo, Katsunori; Ogawa, Taisaku; Hayashi, Masahito; Harada, Yoshie; Nishizaka, Takayuki; Kinosita, Kazuhiko.

    In: PLoS One, Vol. 7, No. 4, e34920, 09.04.2012.

    Research output: Contribution to journalArticle

    Yogo, K, Ogawa, T, Hayashi, M, Harada, Y, Nishizaka, T & Kinosita, K 2012, 'Direct observation of strand passage by dna-topoisomerase and its limited processivity', PLoS One, vol. 7, no. 4, e34920. https://doi.org/10.1371/journal.pone.0034920
    Yogo K, Ogawa T, Hayashi M, Harada Y, Nishizaka T, Kinosita K. Direct observation of strand passage by dna-topoisomerase and its limited processivity. PLoS One. 2012 Apr 9;7(4). e34920. https://doi.org/10.1371/journal.pone.0034920
    Yogo, Katsunori ; Ogawa, Taisaku ; Hayashi, Masahito ; Harada, Yoshie ; Nishizaka, Takayuki ; Kinosita, Kazuhiko. / Direct observation of strand passage by dna-topoisomerase and its limited processivity. In: PLoS One. 2012 ; Vol. 7, No. 4.
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