Detection of Covalent DNA-Bound Spo11 and Topoisomerase Complexes

  • Edgar HartsuikerEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 745)


Topoisomerases can release topological stress and resolve DNA catenanes by a DNA strand breakage and re-ligation mechanism. During the lifetime of the DNA break, the topoisomerase remains covalently linked to the DNA and removes itself when the break is re-ligated. While the lifetime of a covalent topoisomerase–DNA complex is usually short, several clinically important cancer drugs kill cancer cells by inhibiting the removal of covalently linked topoisomerases. The topoisomerase-like protein Spo11 is responsible for meiotic double strand break formation. Spo11 is not able to remove itself and is removed by nucleolytic cleavage. This chapter describes a method which allows the reproducible and quantitative detection of proteins covalently bound to the DNA.

Key words

Topoisomerase I topoisomerase II Spo11 Schizosaccharomyces pombe MRN complex Tdp1 


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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.North West Cancer Research Fund Institute, Bangor UniversityBangorUK

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