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Structural basis for recognition and repair of the endogenous mutagen 8-oxoguanine in DNA

Nature volume 403, pages 859–866 (2000)Cite this article

Abstract

Spontaneous oxidation of guanine residues in DNA generates 8-oxoguanine (oxoG). By mispairing with adenine during replication, oxoG gives rise to a G·C → T·A transversion, a frequent somatic mutation in human cancers. The dedicated repair pathway for oxoG centres on 8-oxoguanine DNA glycosylase (hOGG1), an enzyme that recognizes oxoG·C base pairs, catalysing expulsion of the oxoG and cleavage of the DNA backbone. Here we report the X-ray structure of the catalytic core of hOGG1 bound to oxoG·C-containing DNA at 2.1 Å resolution. The structure reveals the mechanistic basis for the recognition and catalytic excision of DNA damage by hOGG1 and by other members of the enzyme superfamily to which it belongs. The structure also provides a rationale for the biochemical effects of inactivating mutations and polymorphisms in hOGG1. One known mutation, R154H, converts hOGG1 to a pro-mutator by relaxing the specificity of the enzyme for the base opposite oxoG.

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Figure 1: The role of hOGG1 in preventing mutagenesis by oxidized guanines in DNA.
Figure 2: Overall structure of the hOGG1–DNA complex.
Figure 3: Sequence/structural alignment of hOGG1 with other HhH–GPD superfamily members.
Figure 4: Contact interface between hOGG1 and oxoG·C-containing DNA.
Figure 5: Electrostatic potential surface representation (GRASP44) of hOGG1 showing the unusually neutral DNA–protein interaction surface.
Figure 6: Recognition of the oxoG lesion and the estranged cytosine.
Figure 7
Figure 8: Enzymatic activity and substrate specificity of R154H mutant hOGG1.

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Acknowledgements

We thank E. Lawson for assisting in DNA synthesis and protein expression; H. Huang, Q. Fan, L. Chen, S. Ealick, A. Lau, S. Liemann, H. Nash and members of the Verdine, S. C. Harrison and D.C. Wiley laboratories for helpful discussions; C. Schafmeister, R. Crouse, M. Przetakiewicz, R. Sweet, C. Metz and the staff of the CHESS A1 and NSLS X12C synchrotron beamlines for invaluable help with data collection; and K. Haushalter and X. Zhang for critical reading of this manuscript. This research was funded by the NIH.

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  1. Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, 02138, Massachusetts, USA

    Steven D. Bruner, Derek P. G. Norman & Gregory L. Verdine

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Correspondence to Gregory L. Verdine.

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Bruner, S., Norman, D. & Verdine, G. Structural basis for recognition and repair of the endogenous mutagen 8-oxoguanine in DNA. Nature 403, 859–866 (2000). https://doi.org/10.1038/35002510

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