Direct analysis of Holliday junction resolving enzyme in a DNA origami nanostructure

Yuki Suzuki, Masayuki Endo, Cristina Cañas, Silvia Ayora, Juan C. Alonso, Hiroshi Sugiyama, Kunio Takeyasu

Research output: Contribution to journalArticlepeer-review

19 Citations (Scopus)

Abstract

Holliday junction (HJ) resolution is a fundamental step for completion of homologous recombination. HJ resolving enzymes (resolvases) distort the junction structure upon binding and prior cleavage, raising the possibility that the reactivity of the enzyme can be affected by a particular geometry and topology at the junction. Here, we employed a DNA origami nano-scaffold in which each arm of a HJ was tethered through the base-pair hybridization, allowing us to make the junction core either flexible or inflexible by adjusting the length of the DNA arms. Both flexible and inflexible junctions bound to Bacillus subtilis RecU HJ resolvase, while only the flexible junction was efficiently resolved into two duplexes by this enzyme. This result indicates the importance of the structural malleability of the junction core for the reaction to proceed. Moreover, cleavage preferences of RecU-mediated reaction were addressed by analyzing morphology of the reaction products.

Original languageEnglish
Pages (from-to)7421-7428
Number of pages8
JournalNucleic acids research
Volume42
Issue number11
DOIs
Publication statusPublished - 2014 Jun 17
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

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