Cellular Repair of Synthetic Analogs of Oxidative DNA Damage Reveals a Key Structure-Activity Relationship of the Cancer-Associated MUTYH DNA Repair Glycosylase.
| Autor: | Conlon SG; Department of Chemistry, University of California, Davis, One Shields Avenue, Davis, California 95616, United States.; Graduate Program in Chemistry and Chemical Biology, University of California, Davis, One Shields Avenue, Davis, California 95616, United States., Khuu C; Department of Chemistry, University of California, Davis, One Shields Avenue, Davis, California 95616, United States.; Biochemistry, Molecular, Cellular and Developmental Biology Graduate Group, University of California, Davis, One Shields Avenue, Davis, California 95616, United States., Trasviña-Arenas CH; Department of Chemistry, University of California, Davis, One Shields Avenue, Davis, California 95616, United States., Xia T; Department of Chemistry, University of California, Davis, One Shields Avenue, Davis, California 95616, United States.; Graduate Program in Chemistry and Chemical Biology, University of California, Davis, One Shields Avenue, Davis, California 95616, United States., Hamm ML; Department of Chemistry, University of Richmond, 410 Westhampton Way, Richmond, Virginia 23173, United States., Raetz AG; Department of Chemistry, University of California, Davis, One Shields Avenue, Davis, California 95616, United States.; Biochemistry, Molecular, Cellular and Developmental Biology Graduate Group, University of California, Davis, One Shields Avenue, Davis, California 95616, United States., David SS; Department of Chemistry, University of California, Davis, One Shields Avenue, Davis, California 95616, United States.; Graduate Program in Chemistry and Chemical Biology, University of California, Davis, One Shields Avenue, Davis, California 95616, United States.; Biochemistry, Molecular, Cellular and Developmental Biology Graduate Group, University of California, Davis, One Shields Avenue, Davis, California 95616, United States. |
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| Jazyk: | angličtina |
| Zdroj: | ACS central science [ACS Cent Sci] 2024 Jan 26; Vol. 10 (2), pp. 291-301. Date of Electronic Publication: 2024 Jan 26 (Print Publication: 2024). |
| DOI: | 10.1021/acscentsci.3c00784 |
| Abstrakt: | The base excision repair glycosylase MUTYH prevents mutations associated with the oxidatively damaged base, 8-oxo-7,8-dihydroguanine (OG), by removing undamaged misincorporated adenines from OG:A mispairs. Defects in OG:A repair in individuals with inherited MUTYH variants are correlated with the colorectal cancer predisposition syndrome known as MUTYH -associated polyposis (MAP). Herein, we reveal key structural features of OG required for efficient repair by human MUTYH using structure-activity relationships (SAR). We developed a GFP-based plasmid reporter assay to define SAR with synthetically generated OG analogs in human cell lines. Cellular repair results were compared with kinetic parameters measured by adenine glycosylase assays in vitro . Our results show substrates lacking the 2-amino group of OG, such as 8OI:A (8OI = 8-oxoinosine), are not repaired in cells, despite being excellent substrates in in vitro adenine glycosylase assays, new evidence that the search and detection steps are critical factors in cellular MUTYH repair functionality. Surprisingly, modification of the O8/N7H of OG, which is the distinguishing feature of OG relative to G, was tolerated in both MUTYH-mediated cellular repair and in vitro adenine glycosylase activity. The lack of sensitivity to alterations at the O8/N7H in the SAR of MUTYH substrates is distinct from previous work with bacterial MutY, indicating that the human enzyme is much less stringent in its lesion verification. Our results imply that the human protein relies almost exclusively on detection of the unique major groove position of the 2-amino group of OG within OG Competing Interests: The authors declare no competing financial interest. (© 2024 The Authors. Published by American Chemical Society.) |
| Databáze: | MEDLINE |
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