Real-time direct observation of single-molecule dna hydrolysis by exonuclease iii

Hirofumi Kurita, Ken Ichi Inaishi, Ken Torii, Madoka Urisu, Michihiko Nakano, Shinji Katsura, Akira Mizuno

Research output: Contribution to journalArticlepeer-review

21 Citations (Scopus)

Abstract

Single-molecule DNA digestion by exonuclease III, which has 3′ to 5′ exonuclease activity, was analyzed using a micro-channel with two-layer laminar flow. First, a DNA-bead complex was optically trapped in one layer in the absence of exonuclease III permitted the DNA to be stretched by the laminar flow. The exonuclease III reaction was initiated by moving the trapped DNA-bead complex to another layer of flow, which contained exonuclease III. As the reaction proceeded, the fluorescently-stained DNA was observed to shorten. The process was photographed; examination of the photographs showed that the DNA molecule shortened in a linear fashion with respect to the reaction time. The digestion rate obtained from the single-molecule experiment was compared to that measured from a bulk experiment and was found to be ca. 28 times higher than the bulk digestion rate.

Original languageEnglish
Pages (from-to)473-480
Number of pages8
JournalJournal of Biomolecular Structure and Dynamics
Volume25
Issue number5
DOIs
Publication statusPublished - Jan 1 2008
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

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