End-labeling and analysis of Spo11-oligonucleotide complexes in Saccharomyces cerevisiae

Methods Mol Biol. 2009:557:183-95. doi: 10.1007/978-1-59745-527-5_12.

Abstract

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.

Publication types

  • Review

MeSH terms

  • Algorithms
  • DNA Breaks, Double-Stranded*
  • DNA Topoisomerases, Type II / analysis
  • DNA Topoisomerases, Type II / metabolism*
  • Endodeoxyribonucleases
  • In Situ Nick-End Labeling / methods*
  • Macromolecular Substances / analysis*
  • Macromolecular Substances / metabolism
  • Models, Biological
  • Oligonucleotides / analysis
  • Oligonucleotides / metabolism*
  • Protein Binding
  • Saccharomyces cerevisiae / chemistry
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / analysis
  • Saccharomyces cerevisiae Proteins / metabolism*

Substances

  • Macromolecular Substances
  • Oligonucleotides
  • Saccharomyces cerevisiae Proteins
  • Endodeoxyribonucleases
  • Spo11 protein, S cerevisiae
  • DNA Topoisomerases, Type II