End-labeling and analysis of Spo11-oligonucleotide complexes in Saccharomyces cerevisiae.
During meiosis Spo11 catalyzes the formation of DNA double-strand breaks, becoming covalently attached to the 5' ends on both sides of the break during this process. Spo11 is removed from the DSB by single-stranded endonucleolytic cleavage flanking the DSB, liberating a short-lived species consisting of Spo11 protein covalently linked to a short oligonucleotide. The method presented here details how to detect these Spo11-oligo complexes in extracts made from meiotic yeast cells.
MRC Genome Damage and Stability Centre, University of Sussex, Falmer, UK.
Matthew J
MJ
Neale
Scott
S
Keeney
D000465:Algorithms; D053903:DNA Breaks, Double-Stranded; D004250:DNA Topoisomerases, Type II / Q000032:analysis / Q000378:metabolism*; D004706:Endodeoxyribonucleases; D020287:In Situ Nick-End Labeling / Q000379:methods*; D046911:Macromolecular Substances / Q000032:analysis* / Q000378:metabolism; D008954:Models, Biological; D009841:Oligonucleotides / Q000032:analysis / Q000378:metabolism*; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis / Q000378:metabolism*; D000099257:Meiotic Recombination Protein SPO11
D004250:DNA Topoisomerases, Type II; D004706:Endodeoxyribonucleases; D046911:Macromolecular Substances; D009841:Oligonucleotides; D029701:Saccharomyces cerevisiae Proteins; C518938:Spo11 protein, S cerevisiae; D000099257:Meiotic Recombination Protein SPO11