Molecular basis for PrimPol recruitment to replication forks by RPA

Thomas A. Guilliam, Nigel Brissett, Aaron Ehlinger, Benjamin A. Keen, Peter Kolesar, Elaine M. Taylor, Laura J. Bailey, Howard D. Lindsay, Walter J. Chazin, Aidan J. Doherty

Research output: Contribution to journalArticle

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 multifunctional 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 PrimPol's 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 PrimPol's mode of recruitment to stalled forks to facilitate repriming and restart.
Original languageEnglish
Article number15222
JournalNature Communications
Volume8
DOIs
Publication statusPublished - 23 May 2017

Bibliographical note

This work is licensed under a Creative Commons Attribution 4.0
International License. The images or other third party material in this
article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material.
To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

Fingerprint Dive into the research topics of 'Molecular basis for PrimPol recruitment to replication forks by RPA'. Together they form a unique fingerprint.

  • Cite this

    Guilliam, T. A., Brissett, N., Ehlinger, A., Keen, B. A., Kolesar, P., Taylor, E. M., Bailey, L. J., Lindsay, H. D., Chazin, W. J., & Doherty, A. J. (2017). Molecular basis for PrimPol recruitment to replication forks by RPA. Nature Communications, 8, [15222 ]. https://doi.org/10.1038/ncomms15222