Molecular basis for PrimPol recruitment to replication forks by RPA

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

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Abstract

DNA damage and secondary structures can act as potent obstacles to the replication machinery. Persistent stalling events lead to genomic instability and therefore numerous cellular tolerance mechanisms exist to complete genome duplication in the presence of such impediments. In addition to translesion synthesis (TLS) polymerases, 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 restart downstream of lesions and secondary structures. Here, we report that PrimPol is recruited to re-prime stalled replication through its interaction with RPA. Using crystallographic and biophysical approaches, we identify that PrimPol possesses two RPA-interacting motifs and identify the key residues required for these interactions. We demonstrate that one of these motifs is critical for PrimPol’s recruitment to stalled replication forks in vivo thus facilitating its role in re-priming DNA synthesis. In addition, biochemical analysis reveals that RPA serves to stimulate the primase activity of PrimPol. Together, these findings provide unprecedented molecular insights into PrimPol’s mode of recruitment to stalled forks that enables it to efficiently re-prime the restart of DNA replication.

Item Type: Journal Article
Journal or Publication Title: Nature Communications
Uncontrolled Keywords: /dk/atira/pure/subjectarea/asjc/3100
Subjects:
Departments: Faculty of Health and Medicine > Medicine
ID Code: 85534
Deposited By: ep_importer_pure
Deposited On: 23 May 2017 12:18
Refereed?: Yes
Published?: Published
Last Modified: 15 Nov 2019 03:06
URI: https://eprints.lancs.ac.uk/id/eprint/85534

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