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TitleCryo-EM structure of a licensed DNA replication origin.
Journal, issue, pagesNat Commun, Vol. 8, Issue 1, Page 2241, Year 2017
Publish dateDec 21, 2017
AuthorsFerdos Abid Ali / Max E Douglas / Julia Locke / Valerie E Pye / Andrea Nans / John F X Diffley / Alessandro Costa /
PubMed AbstractEukaryotic origins of replication are licensed upon loading of the MCM helicase motor onto DNA. ATP hydrolysis by MCM is required for loading and the post-catalytic MCM is an inactive double hexamer ...Eukaryotic origins of replication are licensed upon loading of the MCM helicase motor onto DNA. ATP hydrolysis by MCM is required for loading and the post-catalytic MCM is an inactive double hexamer that encircles duplex DNA. Origin firing depends on MCM engagement of Cdc45 and GINS to form the CMG holo-helicase. CMG assembly requires several steps including MCM phosphorylation by DDK. To understand origin activation, here we have determined the cryo-EM structures of DNA-bound MCM, either unmodified or phosphorylated, and visualize a phospho-dependent MCM element likely important for Cdc45 recruitment. MCM pore loops touch both the Watson and Crick strands, constraining duplex DNA in a bent configuration. By comparing our new MCM-DNA structure with the structure of CMG-DNA, we suggest how the conformational transition from the loaded, post-catalytic MCM to CMG might promote DNA untwisting and melting at the onset of replication.
External linksNat Commun / PubMed:29269875 / PubMed Central
MethodsEM (single particle)
Resolution4.26 - 7.5 Å
Structure data

EMDB-3833:
Cryo-EM structure of a DDK phosphorylated MCM double hexamer
Method: EM (single particle) / Resolution: 4.26 Å

EMDB-3834:
Cryo-EM structure of an unphosphorylated MCM double hexamer
Method: EM (single particle) / Resolution: 4.77 Å

3960

6f0l

EMDB-3960: Cryo-EM Structure of a Licensed DNA Replication Origin
PDB-6f0l: S. cerevisiae MCM double hexamer bound to duplex DNA
Method: EM (single particle) / Resolution: 6.92 Å

EMDB-4164:
Cryo-EM structure of the DNA bound DDK phosphorylated MCM double hexamer
Method: EM (single particle) / Resolution: 7.5 Å

Chemicals

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

Source
  • Saccharomyces cerevisiae (brewer's yeast)
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
  • synthetic construct (others)
KeywordsHYDROLASE / AAA+ Helicase / Nucleoprotein complex / DNA replication / Double hexamer

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