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.
MeSH terms
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Algorithms
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DNA Breaks, Double-Stranded*
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DNA Topoisomerases, Type II / analysis
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DNA Topoisomerases, Type II / metabolism*
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Endodeoxyribonucleases
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In Situ Nick-End Labeling / methods*
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Macromolecular Substances / analysis*
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Macromolecular Substances / metabolism
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Models, Biological
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Oligonucleotides / analysis
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Oligonucleotides / metabolism*
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Protein Binding
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Saccharomyces cerevisiae / chemistry
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins / analysis
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Saccharomyces cerevisiae Proteins / metabolism*
Substances
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Macromolecular Substances
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Oligonucleotides
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Saccharomyces cerevisiae Proteins
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Endodeoxyribonucleases
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Spo11 protein, S cerevisiae
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DNA Topoisomerases, Type II