Detection of covalent DNA-bound Spo11 and topoisomerase complexes

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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
Original languageEnglish
Title of host publicationDNA recombination: Methods and Protocols
PublisherSpringer Science and Business Media LLC
Pages65-77
ISBN (Print)9781617791291
DOIs
Publication statusPublished - 28 Apr 2011
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