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TitleStructural basis for DNA strand separation by a hexameric replicative helicase.
Journal, issue, pagesNucleic Acids Res, Vol. 43, Issue 17, Page 8551-8563, Year 2015
Publish dateSep 30, 2015
AuthorsYuriy Chaban / Jonathan A Stead / Ksenia Ryzhenkova / Fiona Whelan / Ekaterina P Lamber / Alfred Antson / Cyril M Sanders / Elena V Orlova /
PubMed AbstractHexameric helicases are processive DNA unwinding machines but how they engage with a replication fork during unwinding is unknown. Using electron microscopy and single particle analysis we determined ...Hexameric helicases are processive DNA unwinding machines but how they engage with a replication fork during unwinding is unknown. Using electron microscopy and single particle analysis we determined structures of the intact hexameric helicase E1 from papillomavirus and two complexes of E1 bound to a DNA replication fork end-labelled with protein tags. By labelling a DNA replication fork with streptavidin (dsDNA end) and Fab (5' ssDNA) we located the positions of these labels on the helicase surface, showing that at least 10 bp of dsDNA enter the E1 helicase via a side tunnel. In the currently accepted 'steric exclusion' model for dsDNA unwinding, the active 3' ssDNA strand is pulled through a central tunnel of the helicase motor domain as the dsDNA strands are wedged apart outside the protein assembly. Our structural observations together with nuclease footprinting assays indicate otherwise: strand separation is taking place inside E1 in a chamber above the helicase domain and the 5' passive ssDNA strands exits the assembly through a separate tunnel opposite to the dsDNA entry point. Our data therefore suggest an alternative to the current general model for DNA unwinding by hexameric helicases.
External linksNucleic Acids Res / PubMed:26240379 / PubMed Central
MethodsEM (single particle)
Resolution18.0 - 23.0 Å
Structure data

3087

5a9k

EMDB-3087, PDB-5a9k:
Structural basis for DNA strand separation by a hexameric replicative helicase
Method: EM (single particle) / Resolution: 19.0 Å

EMDB-3088:
Structural basis for DNA strand separation by a hexameric replicative helicase
Method: EM (single particle) / Resolution: 23.0 Å

EMDB-3089:
Structural basis for DNA strand separation by a hexameric replicative helicase
Method: EM (single particle) / Resolution: 18.0 Å

EMDB-3090:
Structural basis for DNA strand separation by a hexameric replicative helicase
Method: EM (single particle) / Resolution: 18.0 Å

EMDB-3091:
Structural basis for DNA strand separation by a hexameric replicative helicase
Method: EM (single particle) / Resolution: 20.0 Å

Chemicals

ChemComp-PO4:
PHOSPHATE ION

ChemComp-MG:
Unknown entry

ChemComp-HOH:
WATER

Source
  • bovine papillomavirus
  • Streptomyces avidinii (bacteria)
  • Mus musculus (house mouse)
KeywordsHYDROLASE / DNA REPLICATION FORK

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