Structure-Activity Relationships Reveal Key Features of 8-Oxoguanine: A Mismatch Detection by the MutY Glycosylase

Amelia H. Manlove, Paige L. McKibbin, Emily L. Doyle, Chandrima Majumdar, Michelle L. Hamm, Sheila S. David

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Base excision repair glycosylases locate and remove damaged bases in DNA with remarkable specificity. The MutY glycosylases, unusual for their excision of undamaged adenines mispaired to the oxidized base 8-oxoguanine (OG), must recognize both bases of the mispair in order to prevent promutagenic activity. Moreover, MutY must effectively find OG:A mismatches within the context of highly abundant and structurally similar T:A base pairs. Very little is known about the factors that initiate MutY's interaction with the substrate when it first encounters an intrahelical OG:A mispair, or about the order of recognition checkpoints. Here, we used structure-activity relationships (SAR) to investigate the features that influence the in vitro measured parameters of mismatch affinity and adenine base excision efficiency by E. coli MutY. We also evaluated the impacts of the same substrate alterations on MutY-mediated repair in a cellular context. Our results show that MutY relies strongly on the presence of the OG base and recognizes multiple structural features at different stages of recognition and catalysis to ensure that only inappropriately mispaired adenines are excised. Notably, some OG modifications resulted in more dramatic reductions in cellular repair than in the in vitro kinetic parameters, indicating their importance for initial recognition events needed to locate the mismatch within DNA. Indeed, the initial encounter of MutY with its target base pair may rely on specific interactions with the 2-amino group of OG in the major groove, a feature that distinguishes OG:A from T:A base pairs. These results furthermore suggest that inefficient substrate location in human MutY homologue variants may prove predictive for the early onset colorectal cancer phenotype known as MUTYH-Associated Polyposis, or MAP.

Original languageEnglish (US)
Pages (from-to)2335-2344
Number of pages10
JournalACS Chemical Biology
Volume12
Issue number9
DOIs
StatePublished - Sep 15 2017

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

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    Manlove, A. H., McKibbin, P. L., Doyle, E. L., Majumdar, C., Hamm, M. L., & David, S. S. (2017). Structure-Activity Relationships Reveal Key Features of 8-Oxoguanine: A Mismatch Detection by the MutY Glycosylase. ACS Chemical Biology, 12(9), 2335-2344. https://doi.org/10.1021/acschembio.7b00389