title: Detection of covalent DNA-bound Spo11 and topoisomerase complexes.
abstract: 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.
authors.affiliation: North West Cancer Research Fund Institute, Bangor University, LL57 2UW, Bangor, UK. E.hartsuiker@bangor.ac.uk
authors.forename: Edgar
authors.initials: E
authors.lastname: Hartsuiker
mesh_terms: D053903:DNA Breaks, Double-Stranded; D004264:DNA Topoisomerases, Type I / Q000235:genetics / Q000378:metabolism*; D004250:DNA Topoisomerases, Type II / Q000235:genetics / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000378:metabolism*; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*; D000099257:Meiotic Recombination Protein SPO11
chemical_list: D004264:DNA Topoisomerases, Type I; D004250:DNA Topoisomerases, Type II; D004706:Endodeoxyribonucleases; D029702:Schizosaccharomyces pombe Proteins; D000099257:Meiotic Recombination Protein SPO11