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

Piberger, Ann Liza, Bowry, Akhil, Kelly, Richard D W, Walker, Alexandra K, González-Acosta, Daniel, Bailey, Laura J, Doherty, Aidan J, Méndez, Juan, Morris, Joanna R, Bryant, Helen E and Petermann, Eva (2020) PrimPol-dependent single-stranded gap formation mediates homologous recombination at bulky DNA adducts. Nature Communications, 11 (1). a5863 1-14. ISSN 2041-1723

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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.

Item Type: Article
Keywords: 4-Nitroquinoline-1-oxide, 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide, Benz(a)Anthracenes, Cell Line, DNA Adducts, DNA Primase, DNA, Single-Stranded, DNA-Directed DNA Polymerase, Homologous Recombination, Humans, Multifunctional Enzymes, Quinolones, Rad51 Recombinase, Single Molecule Imaging, Sister Chromatid Exchange
Schools and Departments: School of Life Sciences > Sussex Centre for Genome Damage and Stability
SWORD Depositor: Mx Elements Account
Depositing User: Mx Elements Account
Date Deposited: 25 Aug 2021 07:24
Last Modified: 25 Aug 2021 07:30
URI: http://sro.sussex.ac.uk/id/eprint/101285

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