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Structure of Saccharomyces cerevisiae DNA polymerase epsilon by cryo–electron microscopy

Nature Structural & Molecular Biology volume 13, pages 3543 (2006) | Download Citation

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Abstract

The structure of the multisubunit yeast DNA polymerase ε (Pol ε) was determined to 20-Å resolution using cryo-EM and single-particle image analysis. A globular domain comprising the catalytic Pol2 subunit is flexibly connected to an extended structure formed by subunits Dpb2, Dpb3 and Dpb4. Consistent with the reported involvement of the latter in interaction with nucleic acids, the Dpb portion of the structure directly faces a single cleft in the Pol2 subunit that seems wide enough to accommodate double-stranded DNA. Primer-extension experiments reveal that Pol ε processivity requires a minimum length of primer-template duplex that corresponds to the dimensions of the extended Dpb structure. Together, these observations suggest a mechanism for interaction of Pol ε with DNA that might explain how the structure of the enzyme contributes to its intrinsic processivity.

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Acknowledgements

F.J.A. is a Scholar of the Leukemia and Lymphoma Society of America. N.S. and O.C. were supported by the Kempe foundation. E.J. is supported by grants from The Swedish Research Council, The Swedish Cancer Society and The Medical Faculty of Umeå University. We thank M. Sedova for help with figures of the final Pol ε model and T.A. Kunkel for comments on the primer-extension assays.

Author information

Author notes

    • Iris K Cheung
    •  & Nasim Sabouri

    These authors contributed equally to this work.

Affiliations

  1. Department of Structural Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.

    • Francisco J Asturias
    • , Iris K Cheung
    •  & Daniel Wepplo
  2. Department of Medical Biochemistry and Biophysics, Umeå University, SE-901 87 Umeå, Sweden.

    • Nasim Sabouri
    • , Olga Chilkova
    •  & Erik Johansson

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Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Francisco J Asturias.

Supplementary information

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  1. 1.

    Supplementary Fig. 1

    Determination of single-hit kinetics conditions

  2. 2.

    Supplementary Fig. 2

    Relative motion of Pol epsilon domains

  3. 3.

    Supplementary Table 1

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DOI

https://doi.org/10.1038/nsmb1040

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