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

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.

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Item Type:	Article
Authors/Creators:	Piberger, A.L. Bowry, A. Kelly, R.D.W. Walker, A.K. González-Acosta, D. Bailey, L.J. Doherty, A.J. Méndez, J. Morris, J.R. Bryant, H.E. https://orcid.org/0000-0003-2720-7020 Petermann, E.
Copyright, Publisher and Additional Information:	© The Author(s) 2020. Open Access: This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Keywords:	Cancer; Genomic instability; Mutation
Dates:	Accepted: 15 October 2020 Published (online): 17 November 2020 Published: 17 November 2020
Institution:	The University of Sheffield
Academic Units:	The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) > Department of Human Metabolism (Sheffield) The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) > Department of Oncology (Sheffield)
Funding Information:	Funder Grant number YORKSHIRE CANCER RESEARCH CO/RL
Depositing User:	Symplectic Sheffield
Date Deposited:	23 Nov 2020 16:12
Last Modified:	23 Nov 2020 16:12
Status:	Published
Publisher:	Springer Science and Business Media LLC
Refereed:	Yes
Identification Number:	10.1038/s41467-020-19570-7
Related URLs:	PubMed URL
Open Archives Initiative ID (OAI ID):	oai:eprints.whiterose.ac.uk:168266

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

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