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Solution structure of a viral DNA polymerase X and evidence for a mutagenic function

Nature Structural Biology volume 8, pages 942–946 (2001)Cite this article

Abstract

The African swine fever virus DNA polymerase X (ASFV Pol X or Pol X), the smallest known nucleotide polymerase, has recently been reported to be an extremely low fidelity polymerase that may be involved in strategic mutagenesis of the viral genome. Here we report the solution structure of Pol X. The structure, unique within the realm of nucleotide polymerases, consists of only palm and fingers subdomains. Despite the absence of a thumb subdomain, which is important for DNA binding in other polymerases, we show that Pol X binds DNA with very high affinity. Further structural analyses suggest a novel mode of DNA binding that may contribute to low fidelity synthesis. We also demonstrate that the ASFV DNA ligase is a low fidelity ligase capable of sealing a nick that contains a G-G mismatch. This supports the hypothesis of a virus-encoded, mutagenic base excision repair pathway consisting of a tandem Pol X/ligase mutator.

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Figure 1: NMR solution structure of Pol X and structural comparison with Pol β.
Figure 2: Mapped DNA binding site of Pol X.
Figure 3: Comparison of DNA Binding by Pol X and Pol β.
Figure 4: Fidelity of ASFV DNA ligase.

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Acknowledgements

We thank C. Yuan for assistance in initial NMR experiments and L. Dixon for her gift of the plasmid pLD20 containing an ASFV genomic fragment that included locus NP419L. R. Coleman and R. Perez assisted in the preparation of DNA. This work was supported by a grant from the National Institutes of Health.

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Authors and Affiliations

  1. Department of Chemistry, The Ohio State University, Columbus, 43210, Ohio, USA

    Alexander K. Showalter, Mei-I Su & Ming-Daw Tsai

  2. The Campus Chemical Instrument Center, The Ohio State University, Columbus, 43210, Ohio, USA

    In-Ja L. Byeon & Ming-Daw Tsai

  3. Department of Biochemistry, The Ohio State University, Columbus, 43210, Ohio, USA

    Ming-Daw Tsai

  4. The Ohio State Biochemistry Program, The Ohio State University, Columbus, 43210, Ohio, USA

    Ming-Daw Tsai

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  1. Alexander K. Showalter
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Correspondence to In-Ja L. Byeon or Ming-Daw Tsai.

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Showalter, A., Byeon, IJ., Su, MI. et al. Solution structure of a viral DNA polymerase X and evidence for a mutagenic function. Nat Struct Mol Biol 8, 942–946 (2001). https://doi.org/10.1038/nsb1101-942

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