Marion Schuller
Molecular basis for DarT ADP-ribosylation of a DNA base
Schuller, Marion; Butler, Rachel; Ariza, Antonio; Tromans-Coia, Callum; Jankevicius, Gytis; Claridge, Tim; Kendall, Sharon; Goh, Shan; Stewart, Graham; Ahel, Ivan
Authors
Rachel Butler
Antonio Ariza
Callum Tromans-Coia
Gytis Jankevicius
Tim Claridge
Sharon Kendall
Shan Goh
Graham Stewart
Ivan Ahel
Abstract
ADP-ribosyltransferases (ARTs) utilise NAD+ to catalyse substrate ADP-ribosylation, thereby regulating cellular pathways or contributing to toxin-mediated pathogenicity of bacteria. Reversible ADP-ribosylation has traditionally been considered a protein-specific modification, but recent in vitro studies have suggested nucleic acids as targets. Here, we present evidence that specific reversible DNA ADP-ribosylation on thymidine bases occurs in cellulo through the DarT/DarG toxin/antitoxin system which is found in a variety of bacteria including global pathogens such as Mycobacterium tuberculosis, EPEC and Pseudomonas aeruginosa. We report the first DarT structure which identifies this protein as a diverged member of the PARP family. Moreover, a set of high-resolution structures in ligand-free, pre-and post-reaction states reveals a specialised mechanism of catalysis that includes a key active-site arginine, extending the canonical ART toolkit. Comparison with the well-established DNA-repair protein ADP-ribosylation complex, PARP/HPF1, offers insights into how the DarT class of ARTs evolved into specific DNA-modifying enzymes. Together, the structural and mechanistic data provide unprecedented detail for a PARP family member and contribute to fundamental understanding of nucleic acid ADP-ribosylation. We furthermore show that thymine-linked ADP-ribose DNA adducts reversed by DarG antitoxin, functioning as non-canonical DNA-repair factor, are utilised not only for targeted DNA damage to induce toxicity but also as a signalling strategy for cellular processes. Using M. tuberculosis as an exemplar we show that DarTG regulates growth by DNA ADP-ribosylation at the origin of chromosome replication
Citation
Schuller, M., Butler, R., Ariza, A., Tromans-Coia, C., Jankevicius, G., Claridge, T., Kendall, S., Goh, S., Stewart, G., & Ahel, I. (2021). Molecular basis for DarT ADP-ribosylation of a DNA base. Nature, https://doi.org/10.1038/s41586-021-03825-4
Journal Article Type | Article |
---|---|
Acceptance Date | Jul 15, 2021 |
Publication Date | Aug 18, 2021 |
Deposit Date | Jul 15, 2021 |
Publicly Available Date | Feb 18, 2022 |
Journal | Nature |
Print ISSN | 0028-0836 |
Electronic ISSN | 1476-4687 |
Publisher | Nature Research |
Peer Reviewed | Peer Reviewed |
DOI | https://doi.org/10.1038/s41586-021-03825-4 |
Public URL | https://rvc-repository.worktribe.com/output/1550088 |
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Publisher Licence URL
http://creativecommons.org/licenses/by-nc-nd/3.0/
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