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Structure of the heterodimeric core primase

Nature Structural & Molecular Biology volume 12, pages 1137–1144 (2005)Cite this article

Abstract

Primases are DNA-dependent RNA polymerases that synthesize the oligoribonucleotide primers essential to DNA replication. In archaeal and eukaryotic organisms, the core primase is a heterodimeric enzyme composed of a small and a large subunit. Here we report a crystallographic and biochemical analysis of the core primase from the archaeon Sulfolobus solfataricus. The structure provides the first three-dimensional description of the large subunit and its interaction with the small subunit. The evolutionary conservation of amino acids at the protein-protein interface implies that the observed mode of subunit association is conserved among archaeal and eukaryotic primases. The orientation of the large subunit in the core primase probably excludes its direct involvement in catalysis. Modeling of a DNA-RNA helix together with structure-based site-directed mutagenesis provides insight into the mechanism of template DNA binding and RNA primer synthesis.

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Figure 1: The Sso core primase.
Figure 2: Structure of the Sso core primase.
Figure 3: The PriS-PriL interface of the Sso core primase.
Figure 4: Analysis of the PriS-PriL interface.
Figure 5: The Gly-Ser dipeptide insertion between PriS residues Glu198 and Ile199 weakens the interaction of the primase subunits.
Figure 6: Interaction of the Sso core primase with DNA template and RNA primer.

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Acknowledgements

This research was supported by a Wellcome Trust senior research fellowship award to L.P. and by the Medical Research Council in the laboratory of S.D.B. We thank X.-J. Lu for help with X3DNA.

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Authors and Affiliations

  1. MRC Cancer Cell Unit, Hutchison MRC Research Centre, Hills Road, Cambridge, CB2 2XZ, UK

    Si-Houy Lao-Sirieix & Stephen D Bell

  2. Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge, CB2 1GA, UK

    Ravi K Nookala & Luca Pellegrini

  3. Laboratory of Molecular Biophysics, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK

    Pietro Roversi

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  1. Si-Houy Lao-Sirieix
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Correspondence to Luca Pellegrini.

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Supplementary information

Supplementary Fig. 1

Crystallographic analysis of the Sso core primase (PDF 871 kb)

Supplementary Fig. 2

Structure-based multiple PriS sequence alignment. (PDF 1147 kb)

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Lao-Sirieix, SH., Nookala, R., Roversi, P. et al. Structure of the heterodimeric core primase. Nat Struct Mol Biol 12, 1137–1144 (2005). https://doi.org/10.1038/nsmb1013

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