PrimPol-dependent single-stranded gap formation mediates homologous recombination at bulky DNA adducts

Liza Piberger, Akhil Bowry, Richard Kelly, Alexa Walker, Daniel Gonzalez-Acosta, Laura Bailey, A Doherty, Juan Mendez, Jo Morris, Helen E Bryant, Eva Petermann

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)
109 Downloads (Pure)

Abstract

Stalled replication forks can be restarted and repaired by RAD51-mediated homologous recombination (HR), but HR can also perform post-replicative repair after bypass of the obstacle. Bulky DNA adducts are important replication-blocking lesions, but it is unknown whether they activate HR at stalled forks or behind ongoing forks. Using mainly BPDE-DNA adducts as model lesions, we show that HR induced by bulky adducts in mammalian cells predominantly occurs at post-replicative gaps formed by the DNA/RNA primase PrimPol. RAD51 recruitment under these conditions does not result from fork stalling, but rather occurs at gaps formed by PrimPol re-priming and resection by MRE11 and EXO1. In contrast, RAD51 loading at double-strand breaks does not require PrimPol. At bulky adducts, PrimPol promotes sister chromatid exchange and genetic recombination. Our data support that HR at bulky adducts in mammalian cells involves post-replicative gap repair and define a role for PrimPol in HR-mediated DNA damage tolerance.
Original languageEnglish
Article number5863
JournalNature Communications
Volume11
Issue number1
DOIs
Publication statusPublished - 17 Nov 2020

Keywords

  • 4-Nitroquinoline-1-oxide/toxicity
  • 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide/metabolism
  • Benz(a)Anthracenes/administration & dosage
  • Cell Line
  • DNA Adducts/genetics
  • DNA Primase/genetics
  • DNA, Single-Stranded
  • DNA-Directed DNA Polymerase/genetics
  • Homologous Recombination/physiology
  • Humans
  • Multifunctional Enzymes/genetics
  • Quinolones/toxicity
  • Rad51 Recombinase/genetics
  • Single Molecule Imaging
  • Sister Chromatid Exchange

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