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Crystal structure of a DNA-dependent RNA polymerase (DNA primase)

Nature Structural Biology volume 8, pages 57–61 (2001)Cite this article

Abstract

Primases are essential components of the DNA replication apparatus in every organism. They catalyze the synthesis of oligoribonucleotides on single-stranded DNA, which subsequently serve as primers for the replicative DNA polymerases. In contrast to bacterial primases, the archaeal enzymes are closely related to their eukaryotic counterparts. We have solved the crystal structure of the catalytic primase subunit from the hyperthermophilic archaeon Pyrococcus furiosus at 2.3 Å resolution by multiwavelength anomalous dispersion methods. The structure shows a two-domain arrangement with a novel zinc knuckle motif located in the primase (prim) domain. In this first structure of a complete protein of the archaeal/eukaryotic primase family, the arrangement of the catalytically active residues resembles the active sites of various DNA polymerases that are unrelated in fold.

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Figure 1: Structure based sequence alignment, activity and Cα trace of Pfu-prim.
Figure 2: Structural overview, surface properties and zinc binding region of Pfu-prim.
Figure 3: Active site topology and hypothetical model of Pfu-prim bound to a DNA/RNA hybrid.

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Acknowledgements

We acknowledge the assistance of H.-D. Bartunik and G.P. Bourenkov at DESY, Hamburg. We thank J. Richardson and S. Steinbacher for helpful discussions.

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  1. Abteilung für Strukturforschung, Max-Planck-Institut für Biochemie, Am Klopferspitz 18A, Martinsried, D-82152 , Germany

    Martin A. Augustin, Robert Huber & Jens T. Kaiser

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  1. Martin A. Augustin
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  2. Robert Huber
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  3. Jens T. Kaiser
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Correspondence to Martin A. Augustin.

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Augustin, M., Huber, R. & Kaiser, J. Crystal structure of a DNA-dependent RNA polymerase (DNA primase). Nat Struct Mol Biol 8, 57–61 (2001). https://doi.org/10.1038/83060

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