Genome-wide mapping of sister chromatid exchange events in single yeast cells using Strand-seq

Clemence Claussin, David Porubsky, Diana C. J. Spierings, Nancy Halsema, Stefan Rentas, Victor Guryev, Peter M. Lansdorp, Michael Chang

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Abstract

Homologous recombination involving sister chromatids is the most accurate, and thus most frequently used, form of recombination-mediated DNA repair. Despite its importance, sister chromatid recombination is not easily studied because it does not result in a change in DNA sequence, making recombination between sister chromatids difficult to detect. We have previously developed a novel DNA template strand sequencing technique, called Strand-seq, that can be used to map sister chromatid exchange (SCE) events genome-wide in single cells. An increase in the rate of SCE is an indicator of elevated recombination activity and of genome instability, which is a hallmark of cancer. In this study, we have adapted Strand-seq to detect SCE in the yeast Saccharomyces cerevisiae. We provide the first quantifiable evidence that most spontaneous SCE events in wild-type cells are not due to the repair of DNA double-strand breaks.

Original languageEnglish
Article number30560
Number of pages17
JournaleLife
Volume6
DOIs
Publication statusPublished - 12-Dec-2017

Keywords

  • REPLICATION PROTEIN-A
  • CONSERVATIVE DNA-SYNTHESIS
  • SACCHAROMYCES-CEREVISIAE
  • BREAK REPAIR
  • HOMOLOGOUS RECOMBINATION
  • MITOTIC RECOMBINATION
  • RAD52 PROTEIN
  • GENE CONVERSION
  • INVERTED-REPEAT
  • CROSSING-OVER

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