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TitleStructural basis for the increased processivity of D-family DNA polymerases in complex with PCNA.
Journal, issue, pagesNat Commun, Vol. 11, Issue 1, Page 1591, Year 2020
Publish dateMar 27, 2020
AuthorsClément Madru / Ghislaine Henneke / Pierre Raia / Inès Hugonneau-Beaufet / Gérard Pehau-Arnaudet / Patrick England / Erik Lindahl / Marc Delarue / Marta Carroni / Ludovic Sauguet /
PubMed AbstractReplicative DNA polymerases (DNAPs) have evolved the ability to copy the genome with high processivity and fidelity. In Eukarya and Archaea, the processivity of replicative DNAPs is greatly enhanced ...Replicative DNA polymerases (DNAPs) have evolved the ability to copy the genome with high processivity and fidelity. In Eukarya and Archaea, the processivity of replicative DNAPs is greatly enhanced by its binding to the proliferative cell nuclear antigen (PCNA) that encircles the DNA. We determined the cryo-EM structure of the DNA-bound PolD-PCNA complex from Pyrococcus abyssi at 3.77 Å. Using an integrative structural biology approach - combining cryo-EM, X-ray crystallography, protein-protein interaction measurements, and activity assays - we describe the molecular basis for the interaction and cooperativity between a replicative DNAP and PCNA. PolD recruits PCNA via a complex mechanism, which requires two different PIP-boxes. We infer that the second PIP-box, which is shared with the eukaryotic Polα replicative DNAP, plays a dual role in binding either PCNA or primase, and could be a master switch between an initiation and a processive phase during replication.
External linksNat Commun / PubMed:32221299 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2.3 - 3.77 Å
Structure data

10401

6t8h

EMDB-10401, PDB-6t8h:
Cryo-EM structure of the DNA-bound PolD-PCNA processive complex from P. abyssi
Method: EM (single particle) / Resolution: 3.77 Å

PDB-6t7x:
Crystal structure of PCNA from P. abyssi
Method: X-RAY DIFFRACTION / Resolution: 2.3 Å

PDB-6t7y:
Structure of PCNA bound to cPIP motif of DP2 from P. abyssi
Method: X-RAY DIFFRACTION / Resolution: 2.7 Å

Chemicals

ChemComp-HOH:
WATER / Water

ChemComp-FE:
Unknown entry / Iron

ChemComp-ZN:
Unknown entry

Source
  • pyrococcus abyssi (archaea)
  • synthetic construct (others)
  • pyrococcus abyssi (strain ge5 / orsay) (archaea)
KeywordsREPLICATION / PCNA / Pyrococcus abyssi / sliding-clamp / DP2 / PolD / PIP-box / DP1

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