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Topoisomerase I-Mediated DNA Relaxation as a Tool to Study Intercalation of Small Molecules into Supercoiled DNA

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Drug-DNA Interaction Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 613))

Abstract

Several biochemical and biophysical methods are available to study the intercalation of a small molecule between two consecutive base pairs of DNA. Among them, the topoisomerase I-mediated DNA relaxation assay has proved highly efficient, relatively easy to handle and very informative to investigate drug binding to DNA. The test relies on the use of a supercoiled plasmid to mimic the topological constraints of genomic DNA. The three main components of the assay - the topoisomerase I enzyme, DNA helix and intercalating small molecules - are presented here in a structural context. The principle of the assay is described in detail, along with a typical experimental protocol.

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Acknowledgements

M.-H.D.-C. thanks the Institut pour la Recherche sur le Cancer (IRCL), Association pour la Recherche contre le Cancer (ARC) and the Ligue Nationale contre le Cancer (Comité du Nord) for grants. P.P. thanks the Institut pour la Recherche sur le Cancer sur le Cancer de Lille (IRCL), the Conseil Régional Nord-Pas-de-Calais and the ARC for a PhD fellowship. The authors are grateful to Sabine Depauw for her technical expertise.

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Authors and Affiliations

  1. INSERM U-837, Jean-Pierre Aubert Research Center (JPARC), Institut de Recherches sur le Cancer de Lille, Lille, France

    Paul Peixoto & Marie-Hélène David-Cordonnier

  2. IMPRT-IFR114, Lille, France

    Paul Peixoto, Christian Bailly & Marie-Hélène David-Cordonnier

  3. Jean-Pierre Aubert Research Center (JPARC), Institut de Recherches sur le Cancer de Lille, Lille, France

    Christian Bailly

  4. Pierre Fabre Research Institute, Lille, France

    Christian Bailly

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  1. Paul Peixoto
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  2. Christian Bailly
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  3. Marie-Hélène David-Cordonnier
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Correspondence to Christian Bailly .

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Editors and Affiliations

  1. School of Biological Sciences, University of Southampton, Southampton, SO17 1BJ, United Kingdom

    Keith R. Fox

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Peixoto, P., Bailly, C., David-Cordonnier, MH. (2010). Topoisomerase I-Mediated DNA Relaxation as a Tool to Study Intercalation of Small Molecules into Supercoiled DNA. In: Fox, K. (eds) Drug-DNA Interaction Protocols. Methods in Molecular Biology, vol 613. Humana Press. https://doi.org/10.1007/978-1-60327-418-0_15

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  • DOI: https://doi.org/10.1007/978-1-60327-418-0_15

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