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Molecular basis for PrimPol recruitment to replication forks by RPA

DOI: 10.1038/ncomms15222 DOI Help

Authors: Thomas A. Guilliam (University of Sussex) , Nigel C. Brissett (University of Sussex) , Aaron Ehlinger (Vanderbilt University School of Medicine) , Benjamin A. Keen (University of Sussex) , Peter Kolesar (University of Sussex) , Elaine M. Taylor (Lancaster University) , Laura J. Bailey (University of Sussex) , Howard D. Lindsay (Lancaster University) , Walter J. Chazin (Vanderbilt University School of Medicine) , Aidan J. Doherty (University of Sussex)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Communications , VOL 8

State: Published (Approved)
Published: May 2017
Diamond Proposal Number(s): 10088

Open Access Open Access

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.

Journal Keywords: DNA replication; Replisome; X-ray crystallography

Diamond Keywords: Enzymes

Subject Areas: Biology and Bio-materials, Chemistry

Instruments: I03-Macromolecular Crystallography

Added On: 07/06/2017 10:16


Discipline Tags:

Life Sciences & Biotech Biophysics Structural biology Chemistry Biochemistry

Technical Tags:

Diffraction Macromolecular Crystallography (MX)