Direct Analysis of Holliday Junction Resolving Enzyme in a DNA Origami Nanostructure

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2014 May 6

PMID 24792171

Citations 10

Authors

Yuki Suzuki

Masayuki Endo

Cristina Canas

Silvia Ayora

Juan C Alonso

Hiroshi Sugiyama

Kunio Takeyasu

Affiliations

Soon will be listed here.

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.

Citing Articles

Minimizing Structural Heterogeneity in DNA Self-Assembled Dye Templating via DNA Origami-Tuned Conformations.

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Structural characterization of human protein NCYM and its complex with a newly identified DNA aptamer using atomic force microscopy and small-angle X-ray scattering.

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Processing of stalled replication forks in Bacillus subtilis.

Carrasco B, Torres R, Moreno-Del Alamo M, Ramos C, Ayora S, Alonso J FEMS Microbiol Rev. 2023; 48(1).

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Dubrovin E Biophys Rev. 2023; 15(5):1015-1033.

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DisA Restrains the Processing and Cleavage of Reversed Replication Forks by the RuvAB-RecU Resolvasome.

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