Structure of human MUTYH and functional profiling of cancer-associated variants reveal an allosteric network between its [4Fe-4S] cluster cofactor and active site required for DNA repair
MUTYH is a clinically important DNA glycosylase that thwarts mutations by initiating base-excision repair at 8-oxoguanine (OG):A lesions. The roles for its [4Fe-4S] cofactor in DNA repair remain enigmatic. Functional profiling of cancer-associated variants near the [4Fe-4S] cofactor reveals that most variations abrogate both retention of the cofactor and enzyme activity. Surprisingly, R241Q and N238S retained the metal cluster and bound substrate DNA tightly, but were completely inactive. We determine the crystal structure of human MUTYH bound to a transition state mimic and this shows that Arg241 and Asn238 build an H-bond network connecting the [4Fe-4S] cluster to the catalytic Asp236 that mediates base excision. The structure of the bacterial MutY variant R149Q, along with molecular dynamics simulations of the human enzyme, support a model in which the cofactor functions to position and activate the catalytic Asp. These results suggest that allosteric cross-talk between the DNA binding [4Fe-4S] cofactor and the base excision site of MUTYH regulate its DNA repair function.
Authors
- Merve Demir (ORCID: https://orcid.org/0000-0002-5721-8268)
- Nuria Tamayo (ORCID: https://orcid.org/0000-0002-0497-5230)
- Carlos H. Trasviña‐Arenas (ORCID: https://orcid.org/0000-0002-9881-375X)
- Martin P. Horvath (ORCID: https://orcid.org/0000-0003-3057-9722)
- G. Andrés Cisneros (ORCID: https://orcid.org/0000-0001-6629-3430)
- Sheila S. David (ORCID: https://orcid.org/0000-0001-5873-7935)
- Mohammad Hashemian (ORCID: https://orcid.org/0000-0003-3891-4126)
- Nallely Hoyos-Gonzalez
- Andrew J. Fisher (ORCID: https://orcid.org/0000-0003-3488-6594)
- Upeksha C. Dissanayake (ORCID: https://orcid.org/0000-0002-6991-3452)
- Wen-Jen Lin
Institutions
- The University of Texas at Dallas (US)
- University of Utah (US)
- University of California System (US)
- Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (MX)
- University of California, Davis (US)
Publication Details
- Journal
- Nature Communications
- Published
- 2025-04-15
- DOI
- https://doi.org/10.1038/s41467-025-58361-w
- Citations
- 12
- Primary Topic
- DNA Repair Mechanisms
- Type
- article
- Field-Weighted Citation Impact
- 9.67
Funders
- National Science Foundation
- U.S. Department of Energy
- University of North Texas
- Consejo Nacional de Ciencia y Tecnología
- National Institutes of Health
- Office of Science
- National Cancer Institute
- National Institute of General Medical Sciences
- Division of Chemistry
- Office of Research Infrastructure Programs, National Institutes of Health
- Argonne National Laboratory