Abstract
Most spontaneous damage to bases in DNA is corrected through the action of the base-excision DNA repair pathway. Base excision repair is initiated by DNA glycosylases, lesion-specific enzymes that intercept aberrant bases in DNA and catalyze their excision. How such proteins accomplish the feat of catalyzing no fewer than five sequential reaction steps using a single active site has been unknown. To help answer this, we report the structure of a trapped catalytic intermediate in DNA repair by human 8-oxoguanine DNA glycosylase. This structure and supporting biochemical results reveal that the enzyme sequesters the excised lesion base and exploits it as a cofactor to participate in catalysis. To our knowledge, the present example represents the first documented case of product-assisted catalysis in an enzyme-catalyzed reaction.
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Acknowledgements
Use of the Argonne National Laboratory Structural Biology Center beamlines at the Advanced Photon Source was supported by the U.S. Department of Energy, Office of Biological and Environmental Research. J.C.F. was supported by an NIH BCMB training grant and by a Graduate Research Fellowship from the NSF. S.D.B is an Eli Lilly Fellow. We are grateful to D.P.G. Norman for assistance with data collection and processing and for helpful discussions. We thank C. Heaton and the entire MacCHESS staff for assistance with data collection. R. Sanishvili, S. Korolev, S. Ginell and A. Joachimak of SBC-CAT at the Advanced Photon Source, Argonne National Laboratory provided valuable advice and assistance.
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Fromme, J., Bruner, S., Yang, W. et al. Product-assisted catalysis in base-excision DNA repair. Nat Struct Mol Biol 10, 204–211 (2003). https://doi.org/10.1038/nsb902
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DOI: https://doi.org/10.1038/nsb902
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