5N8A

Structure of RPA70N in complex with PrimPol (fragment 480-560)

Summary for 5N8A

• Entry DOI: 10.2210/pdb5n8a/pdb
• Related: 5N85
• Descriptor: DNA-directed primase/polymerase protein, Replication protein A 70 kDa DNA-binding subunit (3 entities in total)
• Functional Keywords: complex, replication, basic cleft, protein binding
• Biological source: Homo sapiens (Human); More
• Cellular location: Nucleus: Q96LW4 P27694
• Total number of polymer chains: 2
• Total formula weight: 24515.37

Authors

Brissett, N.C.,Doherty, A.J. (deposition date: 2017-02-23, release date: 2017-06-07, Last modification date: 2024-01-17)

Primary citation

Guilliam, T.A.,Brissett, N.C.,Ehlinger, A.,Keen, B.A.,Kolesar, P.,Taylor, E.M.,Bailey, L.J.,Lindsay, H.D.,Chazin, W.J.,Doherty, A.J.
Molecular basis for PrimPol recruitment to replication forks by RPA.
Nat Commun, 8:15222-15222, 2017

PubMed 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.

PubMed: 28534480
DOI: 10.1038/ncomms15222

Experimental method

X-RAY DIFFRACTION (1.28 Å)