Molecular basis for PrimPol recruitment to replication forks by RPA

Guilliam, Thomas A, Brissett, Nigel C, Ehlinger, Aaron, Keen, Benjamin A, Kolesar, Peter, Taylor, Elaine, Bailey, Laura J, Lindsay, Howard, Chazin, Walter J and Doherty, Aidan J (2017) Molecular basis for PrimPol recruitment to replication forks by RPA. Nature Communications, 8. a15222. ISSN 2041-1723

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Abstract

DNA damage and secondary structures can stall the replication machinery. Cells possess numerous tolerance mechanisms to complete genome duplication in the presence of such impediments. In addition to translesion synthesis (TLS) polymerases, most eukaryotic cells contain a multi-functional replicative enzyme called Primase-Polymerase (PrimPol) that is capable of directly bypassing DNA damage by TLS, as well as repriming replication downstream of impediments. Here, we report that PrimPol is recruited to reprime through its interaction with RPA. Using biophysical and crystallographic approaches, we identify that PrimPol possesses two RPA-binding motifs and ascertained the key residues required for these interactions. We demonstrate that one of these motifs is critical for PrimPolʼs recruitment to stalled replication forks in vivo. In addition, biochemical analysis reveals that RPA serves to stimulate the primase activity of PrimPol. Together, these findings provide significant molecular insights into PrimPolʼs mode of recruitment to stalled forks to facilitate repriming and restart.

Item Type: Article
Keywords: DNA replication; stalling; PrimPol; RPA; priming
Schools and Departments: School of Life Sciences > Sussex Centre for Genome Damage and Stability
Research Centres and Groups: Genome Damage and Stability Centre
Depositing User: Aidan Doherty
Date Deposited: 22 May 2017 11:59
Last Modified: 12 Sep 2017 12:03
URI: http://sro.sussex.ac.uk/id/eprint/68177

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Project NameSussex Project NumberFunderFunder Ref
The role of a novel family of eukaryotic DNA polymerases in mitochondrial DNA replicationG0207BBSRC-BIOTECHNOLOGY & BIOLOGICAL SCIENCES RESEARCH COUNCILBB/H019723/1
Understanding the role of PrimPol in damage tolerance during genome replication in eukaryotic cellsG1621BBSRC-BIOTECHNOLOGY & BIOLOGICAL SCIENCES RESEARCH COUNCILBB/M008800/1
Cell cycle regulation of the NHEJ DNA double-strand break repair pathway in eukaryotesG1554BBSRC-BIOTECHNOLOGY & BIOLOGICAL SCIENCES RESEARCH COUNCILBB/M004236/1