Synthesis of Hemicatenanes for the Study of Type IA Topoisomerases.

Lee, Shun-Hsiao; Hsieh, Tao-Shih; Siaw, Grace Ee-Lu

doi:10.1007/978-1-4939-7459-7_11

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Synthesis of Hemicatenanes for the Study of Type IA Topoisomerases.

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Lee, Shun-Hsiao
Biertümpfel, Christian / Molecular Mechanisms of DNA Repair, Max Planck Institute of Biochemistry, Max Planck Society;

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Citation

Lee, S.-H., Hsieh, T.-S., & Siaw, G.-E.-L. (2018). Synthesis of Hemicatenanes for the Study of Type IA Topoisomerases. Methods in Molecular Biology, 1703, 153-159. doi:10.1007/978-1-4939-7459-7_11.

Cite as: https://hdl.handle.net/21.11116/0000-0002-FDAA-2

Abstract

Hemicatenane is a structure that forms when two DNA duplexes are physically linked through a single-stranded crossover. It is proposed to be an intermediate resulting from double Holliday junction (dHJ) dissolution, repair of replication stalled forks and late stage replication. Our previous study has shown that hemicatenane can be synthesized and dissolved in vitro by hyperthermophilic type IA topoisomerases. Here we present the protocol of hemicatenane synthesis and its structure detection by 2D agarose gel electrophoresis. The generated product can be used as a substrate to study the biochemical mechanism of hemicatenane processing reactions.