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Title3D architecture of DNA Pol alpha reveals the functional core of multi-subunit replicative polymerases.
Journal, issue, pagesEMBO J, Vol. 28, Issue 13, Page 1978-1987, Year 2009
Publish dateJul 8, 2009
AuthorsSebastian Klinge / Rafael Núñez-Ramírez / Oscar Llorca / Luca Pellegrini /
PubMed AbstractEukaryotic DNA replication requires the coordinated activity of the multi-subunit DNA polymerases: Pol alpha, Pol delta and Pol epsilon. The conserved catalytic and regulatory B subunits associate in ...Eukaryotic DNA replication requires the coordinated activity of the multi-subunit DNA polymerases: Pol alpha, Pol delta and Pol epsilon. The conserved catalytic and regulatory B subunits associate in a constitutive heterodimer that represents the functional core of all three replicative polymerases. Here, we combine X-ray crystallography and electron microscopy (EM) to describe subunit interaction and 3D architecture of heterodimeric yeast Pol alpha. The crystal structure of the C-terminal domain (CTD) of the catalytic subunit bound to the B subunit illustrates a conserved mechanism of accessory factor recruitment by replicative polymerases. The EM reconstructions of Pol alpha reveal a bilobal shape with separate catalytic and regulatory modules. Docking of the B-CTD complex in the EM reconstruction shows that the B subunit is tethered to the polymerase domain through a structured but flexible linker. Our combined findings provide a structural template for the common functional architecture of the three major replicative DNA polymerases.
External linksEMBO J / PubMed:19494830 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2.5 - 22.9 Å
Structure data

EMDB-1618:
3D reconstruction of heterodimeric yeast Pol alpha using electron microscopy
Method: EM (single particle) / Resolution: 22.9 Å

PDB-3flo:
Crystal structure of the carboxyl-terminal domain of yeast DNA polymerase alpha in complex with its B subunit
Method: X-RAY DIFFRACTION / Resolution: 2.5 Å

Chemicals

ChemComp-SO4:
SULFATE ION / Sulfate

ChemComp-ZN:
Unknown entry

ChemComp-HOH:
WATER / Water

Source
  • saccharomyces cerevisiae (brewer's yeast)
KeywordsTRANSFERASE / Protein-protein complex / phosphoesterase fold / OB fold / Zinc-binding motif / DNA replication / Nucleus / Phosphoprotein / DNA-binding / DNA-directed DNA polymerase / Nucleotidyltransferase

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