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Structure paper

TitleSynergism between CMG helicase and leading strand DNA polymerase at replication fork.
Journal, issue, pagesNat Commun, Vol. 14, Issue 1, Page 5849, Year 2023
Publish dateSep 20, 2023
AuthorsZhichun Xu / Jianrong Feng / Daqi Yu / Yunjing Huo / Xiaohui Ma / Wai Hei Lam / Zheng Liu / Xiang David Li / Toyotaka Ishibashi / Shangyu Dang / Yuanliang Zhai /
PubMed AbstractThe replisome that replicates the eukaryotic genome consists of at least three engines: the Cdc45-MCM-GINS (CMG) helicase that separates duplex DNA at the replication fork and two DNA polymerases, ...The replisome that replicates the eukaryotic genome consists of at least three engines: the Cdc45-MCM-GINS (CMG) helicase that separates duplex DNA at the replication fork and two DNA polymerases, one on each strand, that replicate the unwound DNA. Here, we determined a series of cryo-electron microscopy structures of a yeast replisome comprising CMG, leading-strand polymerase Polε and three accessory factors on a forked DNA. In these structures, Polε engages or disengages with the motor domains of the CMG by occupying two alternative positions, which closely correlate with the rotational movement of the single-stranded DNA around the MCM pore. During this process, the polymerase remains stably coupled to the helicase using Psf1 as a hinge. This synergism is modulated by a concerted rearrangement of ATPase sites to drive DNA translocation. The Polε-MCM coupling is not only required for CMG formation to initiate DNA replication but also facilitates the leading-strand DNA synthesis mediated by Polε. Our study elucidates a mechanism intrinsic to the replisome that coordinates the activities of CMG and Polε to negotiate any roadblocks, DNA damage, and epigenetic marks encountered during translocation along replication forks.
External linksNat Commun / PubMed:37730685 / PubMed Central
MethodsEM (single particle)
Resolution3.07 - 7.39 Å
Structure data

EMDB-37211, PDB-8kg6:
Yeast replisome in state I
Method: EM (single particle) / Resolution: 3.07 Å

EMDB-37213, PDB-8kg8:
Yeast replisome in state II
Method: EM (single particle) / Resolution: 4.23 Å

EMDB-37215, PDB-8kg9:
Yeast replisome in state III
Method: EM (single particle) / Resolution: 4.52 Å

EMDB-37343, PDB-8w7m:
Yeast replisome in state V
Method: EM (single particle) / Resolution: 4.12 Å

EMDB-37345, PDB-8w7s:
Yeast replisome in state IV
Method: EM (single particle) / Resolution: 7.39 Å

Chemicals

ChemComp-ZN:
Unknown entry

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

ChemComp-MG:
Unknown entry

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

Source
  • saccharomyces cerevisiae (brewer's yeast)
  • saccharomyces cerevisiae s288c (yeast)
KeywordsREPLICATION / replisome / complex / DNA replication

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