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TitleMechanism of PCNA loading by Ctf18-RFC for leading-strand DNA synthesis.
Journal, issue, pagesScience, Vol. 385, Issue 6708, Page eadk5901, Year 2024
Publish dateAug 2, 2024
AuthorsZuanning Yuan / Roxana Georgescu / Nina Y Yao / Olga Yurieva / Michael E O'Donnell / Huilin Li /
PubMed AbstractThe proliferating cell nuclear antigen (PCNA) clamp encircles DNA to hold DNA polymerases (Pols) to DNA for processivity. The Ctf18-RFC PCNA loader, a replication factor C (RFC) variant, is specific ...The proliferating cell nuclear antigen (PCNA) clamp encircles DNA to hold DNA polymerases (Pols) to DNA for processivity. The Ctf18-RFC PCNA loader, a replication factor C (RFC) variant, is specific to the leading-strand Pol (Polε). We reveal here the underlying mechanism of Ctf18-RFC specificity to Polε using cryo-electron microscopy and biochemical studies. We found that both Ctf18-RFC and Polε contain specific structural features that direct PCNA loading onto DNA. Unlike other clamp loaders, Ctf18-RFC has a disordered ATPase associated with a diverse cellular activities (AAA+) motor that requires Polε to bind and stabilize it for efficient PCNA loading. In addition, Ctf18-RFC can pry prebound Polε off of DNA, then load PCNA onto DNA and transfer the PCNA-DNA back to Polε. These elements in both Ctf18-RFC and Polε provide specificity in loading PCNA onto DNA for Polε.
External linksScience / PubMed:39088616 / PubMed Central
MethodsEM (single particle)
Resolution3.2 - 4.1 Å
Structure data

EMDB-41661: Cryo-EM structure of S. cerevisiae Ctf18-RFC-PCNA complex in Apo state conformation I
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-41662: Cryo-EM structure of S. cerevisiae Ctf18-RFC-PCNA complex in Apo state conformation I
Method: EM (single particle) / Resolution: 3.5 Å

41663

8tw9

EMDB-41663, PDB-8tw9:
Cryo-EM structure of S. cerevisiae PolE-Ctf18-8-1-DNA
Method: EM (single particle) / Resolution: 3.6 Å

41664

8twa

EMDB-41664, PDB-8twa:
Cryo-EM structure of S. cerevisiae Ctf18-RFC-PCNA-PolE-DNA complex
Method: EM (single particle) / Resolution: 4.1 Å

EMDB-41665: Cryo-EM structure of S. cerevisiae Ctf18-RFC-PCNA-DNA complex
Method: EM (single particle) / Resolution: 3.2 Å

44356

9b8r

EMDB-44356, PDB-9b8r:
Cryo-EM structure of S. cerevisiae PolE-core-DNA
Method: EM (single particle) / Resolution: 3.5 Å

Chemicals

ChemComp-SF4:
IRON/SULFUR CLUSTER

ChemComp-AGS:
PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER / ATP-gamma-S, energy-carrying molecule analogue*YM

ChemComp-MG:
Unknown entry

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

Source
  • saccharomyces cerevisiae (brewer's yeast)
  • synthetic construct (others)
  • dna molecule (others)
KeywordsDNA BINDING PROTEIN/DNA / Pol2 / Ctf18-8-1 / DNA / DNA BINDING PROTEIN-DNA complex / Ctf18 / RFC2-5 / PCNA / REPLICATION

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