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TitleThe dynamic nature of the human origin recognition complex revealed through five cryoEM structures.
Journal, issue, pagesElife, Vol. 9, Year 2020
Publish dateAug 18, 2020
AuthorsMatt J Jaremko / Kin Fan On / Dennis R Thomas / Bruce Stillman / Leemor Joshua-Tor /
PubMed AbstractGenome replication is initiated from specific origin sites established by dynamic events. The Origin Recognition Complex (ORC) is necessary for orchestrating the initiation process by binding to ...Genome replication is initiated from specific origin sites established by dynamic events. The Origin Recognition Complex (ORC) is necessary for orchestrating the initiation process by binding to origin DNA, recruiting CDC6, and assembling the MCM replicative helicase on DNA. Here we report five cryoEM structures of the human ORC (HsORC) that illustrate the native flexibility of the complex. The absence of ORC1 revealed a compact, stable complex of ORC2-5. Introduction of ORC1 opens the complex into several dynamic conformations. Two structures revealed dynamic movements of the ORC1 AAA+ and ORC2 winged-helix domains that likely impact DNA incorporation into the ORC core. Additional twist and pinch motions were observed in an open ORC conformation revealing a hinge at the ORC5·ORC3 interface that may facilitate ORC binding to DNA. Finally, a structure of ORC was determined with endogenous DNA bound in the core revealing important differences between human and yeast origin recognition.
External linksElife / PubMed:32808929 / PubMed Central
MethodsEM (single particle)
Resolution3.2 - 4.4 Å
Structure data

22417

7jpo

EMDB-22417, PDB-7jpo:
ORC-O1AAA: Human Origin Recognition Complex (ORC) with dynamic/unresolved ORC2 WH
Method: EM (single particle) / Resolution: 3.2 Å

22418

7jpp

EMDB-22418, PDB-7jpp:
ORC-O2WH: Human Origin Recognition Complex (ORC) with dynamic/unresolved ORC1 AAA+ domain
Method: EM (single particle) / Resolution: 3.7 Å

22419

7jpq

EMDB-22419, PDB-7jpq:
ORC-O2-5: Human Origin Recognition Complex (ORC) with subunits 2,3,4,5
Method: EM (single particle) / Resolution: 3.5 Å

22420

7jpr

EMDB-22420, PDB-7jpr:
ORC-OPEN: Human Origin Recognition Complex (ORC) in an open conformation
Method: EM (single particle) / Resolution: 4.0 Å

22421

7jps

EMDB-22421, PDB-7jps:
ORC-DNA: Human Origin Recognition Complex (ORC) with DNA bound in the core
Method: EM (single particle) / Resolution: 4.4 Å

Chemicals

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

ChemComp-MG:
Unknown entry

ChemComp-K:
Unknown entry

Source
  • homo sapiens (human)
  • spodoptera frugiperda (fall armyworm)
KeywordsREPLICATION / AAA+ / ORC / DNA-binding / cryoEM / replication/DNA / replication-DNA complex

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