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TitleMolecular Basis for ATP-Hydrolysis-Driven DNA Translocation by the CMG Helicase of the Eukaryotic Replisome.
Journal, issue, pagesCell Rep, Vol. 28, Issue 10, Page 2673-22688.e8, Year 2019
Publish dateSep 3, 2019
AuthorsPatrik Eickhoff / Hazal B Kose / Fabrizio Martino / Tatjana Petojevic / Ferdos Abid Ali / Julia Locke / Nele Tamberg / Andrea Nans / James M Berger / Michael R Botchan / Hasan Yardimci / Alessandro Costa /
PubMed AbstractIn the eukaryotic replisome, DNA unwinding by the Cdc45-MCM-Go-Ichi-Ni-San (GINS) (CMG) helicase requires a hexameric ring-shaped ATPase named minichromosome maintenance (MCM), which spools single- ...In the eukaryotic replisome, DNA unwinding by the Cdc45-MCM-Go-Ichi-Ni-San (GINS) (CMG) helicase requires a hexameric ring-shaped ATPase named minichromosome maintenance (MCM), which spools single-stranded DNA through its central channel. Not all six ATPase sites are required for unwinding; however, the helicase mechanism is unknown. We imaged ATP-hydrolysis-driven translocation of the CMG using cryo-electron microscopy (cryo-EM) and found that the six MCM subunits engage DNA using four neighboring protomers at a time, with ATP binding promoting DNA engagement. Morphing between different helicase states leads us to suggest a non-symmetric hand-over-hand rotary mechanism, explaining the asymmetric requirements of ATPase function around the MCM ring of the CMG. By imaging of a higher-order replisome assembly, we find that the Mrc1-Csm3-Tof1 fork-stabilization complex strengthens the interaction between parental duplex DNA and the CMG at the fork, which might support the coupling between DNA translocation and fork unwinding.
External linksCell Rep / PubMed:31484077 / PubMed Central
MethodsEM (single particle)
Resolution3.7 - 4.46 Å
Structure data

4785

6raw

EMDB-4785: D. melanogaster CMG-DNA with ATP, State 1A
PDB-6raw: D. melanogaster CMG-DNA, State 1A
Method: EM (single particle) / Resolution: 3.7 Å

4786

6rax

EMDB-4786: D. melanogaster CMG-DNA with ATP, State 1B
PDB-6rax: D. melanogaster CMG-DNA, State 1B
Method: EM (single particle) / Resolution: 3.99 Å

4787

6ray

EMDB-4787: D. melanogaster CMG-DNA with ATP, State 2A
PDB-6ray: D. melanogaster CMG-DNA, State 2A
Method: EM (single particle) / Resolution: 4.28 Å

4788

6raz

EMDB-4788: D. melanogaster CMG-DNA with ATP, State 2B
PDB-6raz: D. melanogaster CMG-DNA, State 2B
Method: EM (single particle) / Resolution: 4.46 Å

Chemicals

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
  • drosophila melanogaster (fruit fly)
KeywordsREPLICATION / Helicase / ATPase / AAA+ / DNA unwinding / HYDOLASE / HYDROLASE

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