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Investigation of Break-Induced Replication in Yeast

  • Beth Osia
  • Rajula Elango
  • Juraj Kramara
  • Steven A. Roberts
  • Anna MalkovaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2153)

Abstract

Repair of double-strand DNA breaks (DSBs) is important for preserving genomic integrity and stability. Break-induced replication (BIR) is a mechanism aimed to repair one-ended double-strand DNA breaks, similar to those formed by replication fork collapse or by telomere erosion. Unlike S-phase replication, BIR is carried out by a migrating DNA bubble and is associated with conservative inheritance of newly synthesized DNA. This unusual DNA synthesis leads to high level of mutagenesis and chromosomal rearrangements during BIR. Here, we focus on several genetic and molecular methods to investigate BIR using our system in yeast Saccharomyces cerevisiae where BIR is initiated by a site-specific DNA break, and the repair involves two copies of chromosome III.

Key words

Break-induced replication Double-strand break Single-stranded DNA Homologous recombination Contour-clamped homogenous electric field electrophoresis Gross chromosomal rearrangements APOBEC 

Notes

Acknowledgments

A.M. is supported by R35GM127006 grant from NIGMS, R03ES029306, and R21 ES030307 from NIEHS. S.A.R. is supported by R01 award CA218112 from the National Cancer Institute.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  • Beth Osia
    • 1
  • Rajula Elango
    • 2
    • 3
  • Juraj Kramara
    • 1
  • Steven A. Roberts
    • 4
  • Anna Malkova
    • 1
    Email author
  1. 1.Department of BiologyUniversity of IowaIowa CityUSA
  2. 2.Department of Medicine, Division of Hematology-Oncology, Cancer Research InstituteHarvard Medical SchoolBostonUSA
  3. 3.Beth Israel Deaconess Medical CenterHarvard Medical SchoolBostonUSA
  4. 4.School of Molecular Biosciences, Center for Reproductive BiologyWashington State UniversityPullmanUSA

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