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TitleThe human pre-replication complex is an open complex.
Journal, issue, pagesCell, Vol. 186, Issue 1, Page 98-111.e21, Year 2023
Publish dateJan 5, 2023
AuthorsJian Li / Jiangqing Dong / Weitao Wang / Daqi Yu / Xinyu Fan / Yan Chit Hui / Clare S K Lee / Wai Hei Lam / Nathan Alary / Yang Yang / Yingyi Zhang / Qian Zhao / Chun-Long Chen / Bik-Kwoon Tye / Shangyu Dang / Yuanliang Zhai /
PubMed AbstractIn eukaryotes, DNA replication initiation requires assembly and activation of the minichromosome maintenance (MCM) 2-7 double hexamer (DH) to melt origin DNA strands. However, the mechanism for this ...In eukaryotes, DNA replication initiation requires assembly and activation of the minichromosome maintenance (MCM) 2-7 double hexamer (DH) to melt origin DNA strands. However, the mechanism for this initial melting is unknown. Here, we report a 2.59-Å cryo-electron microscopy structure of the human MCM-DH (hMCM-DH), also known as the pre-replication complex. In this structure, the hMCM-DH with a constricted central channel untwists and stretches the DNA strands such that almost a half turn of the bound duplex DNA is distorted with 1 base pair completely separated, generating an initial open structure (IOS) at the hexamer junction. Disturbing the IOS inhibits DH formation and replication initiation. Mapping of hMCM-DH footprints indicates that IOSs are distributed across the genome in large clusters aligning well with initiation zones designed for stochastic origin firing. This work unravels an intrinsic mechanism that couples DH formation with initial DNA melting to license replication initiation in human cells.
External linksCell / PubMed:36608662
MethodsEM (single particle)
Resolution2.59 - 2.95 Å
Structure data

32258

7w1y

EMDB-32258, PDB-7w1y:
Human MCM double hexamer bound to natural DNA duplex (polyAT/polyTA)
Method: EM (single particle) / Resolution: 2.59 Å

EMDB-33320: Cryo-EM map of hMCM-DH R195A/L209G mutant
Method: EM (single particle) / Resolution: 2.95 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

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

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

Source
  • homo sapiens (human)
  • Human (human)
KeywordsCELL CYCLE/DNA / replication / CELL CYCLE-DNA complex

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