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Structural basis for removal of adenine mispaired with 8-oxoguanine by MutY adenine DNA glycosylase

A Correction to this article was published on 12 February 2004

Abstract

The genomes of aerobic organisms suffer chronic oxidation of guanine to the genotoxic product 8-oxoguanine (oxoG)1. Replicative DNA polymerases misread oxoG residues and insert adenine instead of cytosine opposite the oxidized base. Both bases in the resulting A·oxoG mispair are mutagenic lesions, and both must undergo base-specific replacement to restore the original C·G pair. Doing so represents a formidable challenge to the DNA repair machinery, because adenine makes up roughly 25% of the bases in most genomes. The evolutionarily conserved enzyme adenine DNA glycosylase (called MutY in bacteria and hMYH in humans) initiates repair of A·oxoG to C·G by removing the inappropriately paired adenine base from the DNA backbone. A central issue concerning MutY function is the mechanism by which A·oxoG mispairs are targeted among the vast excess of A·T pairs. Here we report the use of disulphide crosslinking2 to obtain high-resolution crystal structures of MutY–DNA lesion-recognition complexes. These structures reveal the basis for recognizing both lesions in the A·oxoG pair and for catalysing removal of the adenine base.

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Figure 1: Pathway for oxidation and repair of guanine in DNA.
Figure 2: MutY–DNA complex.
Figure 3: Enzyme–DNA interface and oxoG recognition.
Figure 4: Close-up views of the MutY active site.

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Acknowledgements

We thank H. Nash for the reduced abasic phosphoramidite; Y. Korkhin for assistance with data collection and processing; M. Becker for beamline assistance; S. Bruner and J. J. Miranda for critically reading the manuscript; and Enanta Pharmaceuticals for use of their X-ray generator and detector. Some data for this study were measured at beamline X25 of the National Synchrotron Light Source; financial support for this beamline comes from the NIH and the United States Department of Energy.

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Correspondence to Gregory L. Verdine.

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The authors declare that they have no competing financial interests.

Supplementary information

Supplementary figure 1: Disulfide cross-linking strategy and results. (PDF 164 kb)

Supplementary figure 2: Stereoview of the interactions between the enzyme and oxoG-strand of DNA. (PDF 166 kb)

41586_2004_BFnature02306_MOESM3_ESM.pdf

Supplementary figure 3: MutY Sequence alignment showing the amino acid sequences from humans, the bacterium Escherichia coli (E.co.), the thermophilic bacterium Bacillus stearothermophilus (B.st., the ortholog used in this work), and the fission yeast Schizosaccharomyces pombe (S.po.). (PDF 93 kb)

Supplementary figure 4: Structural and crystallographic results. (PDF 126 kb)

Supplementary figure 5: Substrate and product adenine active site superposition. (PDF 129 kb)

Supplementary table: Crystallographic statistics. (PDF 81 kb)

Supplementary information, methods and figure legends. (PDF 42 kb)

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Fromme, J., Banerjee, A., Huang, S. et al. Structural basis for removal of adenine mispaired with 8-oxoguanine by MutY adenine DNA glycosylase. Nature 427, 652–656 (2004). https://doi.org/10.1038/nature02306

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  • DOI: https://doi.org/10.1038/nature02306

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