AI Article Synopsis
- Holliday junction (HJ) resolution is essential for homologous recombination, and resolvases change the junction's structure, potentially affecting their activity.
- A DNA origami nano-scaffold was used to create HJs with either flexible or inflexible cores by altering DNA arm lengths.
- Only the flexible junction was effectively resolved by the Bacillus subtilis RecU resolvase, highlighting the role of junction structure in the resolution process.
Article 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.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4066755 | PMC |
| http://dx.doi.org/10.1093/nar/gku320 | DOI Listing |
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