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TitleStructure of the pre-initiation complex explains CMGE biogenesis.
Journal, issue, pagesNature, Year 2026
Publish dateFeb 23, 2026 (structure data deposition date)
AuthorsThomas Pühringer / Berta Canal / Giacomo Palm / Agata Butryn / Emma C Couves / Oliver Willhoft / Jacob S Lewis / John F X Diffley / Alessandro Costa /
PubMed AbstractWhen cells enter S phase, bidirectional DNA replication is initiated through the kinase-regulated recruitment of three activators (Cdc45, GINS and Pol ε) to a duplex-DNA-loaded double hexamer of ...When cells enter S phase, bidirectional DNA replication is initiated through the kinase-regulated recruitment of three activators (Cdc45, GINS and Pol ε) to a duplex-DNA-loaded double hexamer of minichromosome maintenance (MCM) ATPases. Together, these proteins form two CMGE helicases that establish divergent replication forks as they become separated. Here, to gain an understanding of CMGE biogenesis, we reconstituted the pre-initiation complex with purified yeast proteins. The cryo-electron-microscopy structure shows a set of firing factors caught in the act of assembling two symmetrical CMGEs. We show how stepwise complex formation reshapes MCM in preparation for DNA opening, and we explain how ATP promotes firing-factor ejection and CMGE maturation. We find that although Sld2 facilitates the recruitment of GINS to MCM, as expected, it also aids the efficient separation of the CMGE dimer, and is essential for the ejection of the lagging strand from MCM. These findings have direct implications for our understanding of the metazoan Sld2 orthologue, RECQL4, and point to a replication-fork establishment mechanism that is conserved across eukaryotes.
External linksNature / PubMed:42310460
MethodsEM (single particle)
Resolution2.7 - 3.7 Å
Structure data

PDB-28vy:
sCMGE assembled on ARS1 DNA with Sld2 and RPA
Method: ELECTRON MICROSCOPY / Resolution: 2.7 Å

PDB-9rhi:
pre-Initiation Complex on ARS1 DNA (monomer)
Method: ELECTRON MICROSCOPY / Resolution: 3.2 Å

PDB-9rhj:
Pre-Initiation Complex on ARS1 DNA (dimer)
Method: ELECTRON MICROSCOPY / Resolution: 3.4 Å

PDB-9rhl:
Phospho-DH bound by Sld3-MBD on ARS1 DNA
Method: ELECTRON MICROSCOPY / Resolution: 3.1 Å

PDB-9rhm:
Phospho-MCM double hexamer bound to Sld3-Sld7-Cdc45 on ARS1 DNA
Method: ELECTRON MICROSCOPY / Resolution: 3.7 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-ZN:
Unknown entry

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

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM

Source
  • saccharomyces cerevisiae (brewer's yeast)
  • synthetic construct (others)
KeywordsREPLICATION / Macromolecular complex / DNA / ATPase / Helicase MCM2-7 / Macromolecular Complex DNA ATPase Helicase MCM2-7 / Helicase / MCM2-7

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