Redox-State-Dependent Structural Changes within a Prokaryotic 6–4 Photolyase
Photolyases repair UV damage to DNA by using absorbed blue light. Within the photolyase/cryptochrome superfamily (PCSf), a major subgroup consists of prokaryotic (6-4) photolyases. These enzymes rely on flavin adenine dinucleotide (FAD) as a catalytic cofactor, besides an ancillary antenna chromophore, and a [4Fe-4S] cluster with yet unknown function. For the prokaryotic 6-4 photolyase of <i>Caulobacter crescentus</i>, we investigated structural changes associated with its different redox states by damage-free crystallography using X-ray free-electron lasers. EPR and optical spectroscopy confirmed redox-dependent structural transitions, including the formation of an oxidized [4Fe-4S]<sup>3+</sup> cluster with the dynamic cleavage of a single iron-sulfur bond. Photoreduction to the catalytic FADH<sup>-</sup> state alters the flavin binding site at the proximal aromatic pair Y390/F394 that is part of the electron transport pathway. Upon oxidation, the observable structural transitions of the protein matrix around the [4Fe-4S] cluster may affect DNA binding and are consistent with the much-debated role of the iron-sulfur cluster in DNA-binding proteins for quenching electron holes.
Authors
- Manuel Maestre‐Reyna (ORCID: https://orcid.org/0000-0002-9566-7216)
- Jungmin Kang (ORCID: https://orcid.org/0000-0002-3506-6400)
- Hans‐Joachim Emmerich (ORCID: https://orcid.org/0000-0002-3260-4509)
- Antoine Royant (ORCID: https://orcid.org/0000-0002-1919-8649)
- Takehiko Tosha (ORCID: https://orcid.org/0000-0002-8971-0759)
- Eriko Nango (ORCID: https://orcid.org/0000-0001-9851-7355)
- Jéssica C. Soares
- Fangjia Luo (ORCID: https://orcid.org/0000-0003-0631-6236)
- Nicolas Caramello (ORCID: https://orcid.org/0000-0003-0025-0213)
- Lars‐Oliver Essen (ORCID: https://orcid.org/0000-0003-4272-4026)
- Mohamed Watad
- Junpei Yamamoto (ORCID: https://orcid.org/0000-0002-5036-991X)
- Sylvain Engilberge (ORCID: https://orcid.org/0000-0001-8680-6790)
- Kensuke Tono (ORCID: https://orcid.org/0000-0003-1218-3759)
- Po‐Hsun Wang (ORCID: https://orcid.org/0000-0001-7359-934X)
- Yuhei Hosokawa (ORCID: https://orcid.org/0000-0001-9559-8567)
- Christian Rosner
- Shigeki Owada (ORCID: https://orcid.org/0000-0002-1451-7612)
- Andrea Bologna (ORCID: https://orcid.org/0009-0007-5375-7057)
- V. FUCHS
- Antonio J. Pierik (ORCID: https://orcid.org/0000-0002-1509-6370)
- Mai Nakamura (ORCID: https://orcid.org/0000-0002-4516-5496)
- Yoshitaka Bessho (ORCID: https://orcid.org/0000-0001-9297-7473)
- Ming‐Daw Tsai (ORCID: https://orcid.org/0000-0003-1374-0414)
Institutions
- Centre National de la Recherche Scientifique (FR)
- University of Kaiserslautern (DE)
- Universität Hamburg (DE)
- Philipps University of Marburg (DE)
- University of Sannio (IT)
- National Taiwan University (TW)
- Tohoku University (JP)
- Commissariat à l'Énergie Atomique et aux Énergies Alternatives (FR)
- European Synchrotron Radiation Facility (FR)
- CEA Grenoble (FR)
- Institute of Biological Chemistry, Academia Sinica (TW)
- Faculty (United Kingdom) (GB)
- SPring-8 (JP)
- Japan Synchrotron Radiation Research Institute (JP)
- Institut de Biologie Structurale (FR)
- Hamburg Centre for Ultrafast Imaging
- The University of Tokyo (JP)
- Université Grenoble Alpes (FR)
- The University of Osaka (JP)
Publication Details
- Journal
- Journal of the American Chemical Society
- Published
- 2025-04-29
- DOI
- https://doi.org/10.1021/jacs.4c18116
- Citations
- 2
- Primary Topic
- Light effects on plants
- Type
- article
- Field-Weighted Citation Impact
- 3.50
Funders
- Deutsche Forschungsgemeinschaft
- Agence Nationale de la Recherche
- Academia Sinica
- Centre National de la Recherche Scientifique
- National Taiwan University
- National Science and Technology Council
- Japan Society for the Promotion of Science
- Air Force Office of Scientific Research