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TitleStructure of a human replisome shows the organisation and interactions of a DNA replication machine.
Journal, issue, pagesEMBO J, Vol. 40, Issue 23, Page e108819, Year 2021
Publish dateDec 1, 2021
AuthorsMorgan L Jones / Yasemin Baris / Martin R G Taylor / Joseph T P Yeeles /
PubMed AbstractThe human replisome is an elaborate arrangement of molecular machines responsible for accurate chromosome replication. At its heart is the CDC45-MCM-GINS (CMG) helicase, which, in addition to ...The human replisome is an elaborate arrangement of molecular machines responsible for accurate chromosome replication. At its heart is the CDC45-MCM-GINS (CMG) helicase, which, in addition to unwinding the parental DNA duplex, arranges many proteins including the leading-strand polymerase Pol ε, together with TIMELESS-TIPIN, CLASPIN and AND-1 that have key and varied roles in maintaining smooth replisome progression. How these proteins are coordinated in the human replisome is poorly understood. We have determined a 3.2 Å cryo-EM structure of a human replisome comprising CMG, Pol ε, TIMELESS-TIPIN, CLASPIN and AND-1 bound to replication fork DNA. The structure permits a detailed understanding of how AND-1, TIMELESS-TIPIN and Pol ε engage CMG, reveals how CLASPIN binds to multiple replisome components and identifies the position of the Pol ε catalytic domain. Furthermore, the intricate network of contacts contributed by MCM subunits and TIMELESS-TIPIN with replication fork DNA suggests a mechanism for strand separation.
External linksEMBO J / PubMed:34694004 / PubMed Central
MethodsEM (single particle)
Resolution3.2 - 4.3 Å
Structure data

EMDB-13375, PDB-7pfo:
Core human replisome
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-13376:
AND-1/CDC45/GINS multibody
Method: EM (single particle) / Resolution: 4.2 Å

EMDB-13377:
Pol-Epsilon/CDC45/GINS multibody refinement
Method: EM (single particle) / Resolution: 4.3 Å

EMDB-13384:
Core human replisome minus CLASPIN
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-13457:
cryoSPARC refinement of core human replisome displaying lagging strand density
Method: EM (single particle) / Resolution: 4.0 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

ChemComp-ANP:
PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER / AMP-PNP, energy-carrying molecule analogue*YM

ChemComp-SO4:
SULFATE ION

Source
  • homo sapiens (human)
KeywordsREPLICATION / DNA replication / helicase / CMG / TIMELESS / TIPIN / CLASPIN / AND-1 / EPSILON / POLYMERASE / REPLISOME

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