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Biochemical and Genetic Analyses of DNA Topoisomerase 1-Mediated DNA Damage

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Camptothecins in Cancer Therapy

Part of the book series: Cancer Drug Discovery and Development ((CDD&D))

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Abstract

Eukaryotic DNA topoisomerase 1 (Top1) is a highly conserved enzyme that catalyzes changes in the linkage of DNA strands (reviewed in refs. 1–3). Such changes in DNA topology are important during cellular processes involving DNA, including DNA replication, recombination, transcription, and chromosome condensation (1–3). The monomeric Top1 enzyme, encoded by the Top1 gene, binds to duplex DNA and catalyzes the transient cleavage and relegation of a single DNA strand. This is achieved by the nucleophilic attack of the active site tyrosine on a DNA phosphodiester bond to generate a phosphotyrosyl linkage between the enzyme and the 3′-end of the nicked DNA. The formation of this enzyme-linked nick allows for the rotation of the noncovalently held DNA end around phosphodiester bonds in the nonscissile strand to effect changes in DNA linking number. In a second transesterification reaction, the 5′OH DNA end attacks the phosphotyrosyl bond to restore the phosphodiester backbone bond and liberate the enzyme. The formation of a covalent Top1p-DNA complex is the hallmark of topoisomerase-catalyzed reactions and acts to conserve the energy of the cleaved DNA bond such that the concerted nicking and relegation of DNA stands does not require an exogenous energy source, such as adenosine triphosphate (ATP). The type IB enzymes, such as eukaryotic Top1, are distinct from type IA and type II enzymes in the formation of a 3′-phosphotyrosyl bond. Recent structural insights suggest mechanistic similarities between type IB enzymes and tyrosine recombinases, such as Cre and Int, which also form a covalent linkage with a 3′-phosphoryl DNA end (4,5).

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Authors and Affiliations

  1. Department of Molecular Pharmacology, St. Jude Children’s Research Hospital, Memphis, TN

    Robert C. A. M. van Waardenburg PhD & Mary-Ann Bjornsti PhD

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  1. Robert C. A. M. van Waardenburg PhD
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Editors and Affiliations

  1. Department of Pharmacy Practice and Science, College of Pharmacy, University of Kentucky, Lexington, KY

    Val R. Adams PharmD

  2. Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY

    Thomas G. Burke PhD

  3. Experimental Therapeutics Program, Lucille P. Markey Cancer Center, University of Kentucky Medical Center, University of Kentucky, Lexington, KY

    Thomas G. Burke PhD

  4. Tigen Pharmaceuticals Inc., Lexington, KY

    Thomas G. Burke PhD

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van Waardenburg, R.C.A.M., Bjornsti, MA. (2005). Biochemical and Genetic Analyses of DNA Topoisomerase 1-Mediated DNA Damage. In: Adams, V.R., Burke, T.G. (eds) Camptothecins in Cancer Therapy. Cancer Drug Discovery and Development. Humana Press. https://doi.org/10.1385/1-59259-866-8:109

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  • DOI: https://doi.org/10.1385/1-59259-866-8:109

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