Cutts, E.E. orcid.org/0000-0003-3290-4293, Saravanan, S. orcid.org/0000-0002-5974-7710, Girvan, P. orcid.org/0000-0003-1411-0094 et al. (4 more authors) (2025) Substrate accessibility regulation of human TopIIa decatenation by cohesin. Nature Communications, 16 (1). 7200. ISSN: 2041-1723
Abstract
Human topoisomerase II alpha (TOP2α) resolves DNA intertwines between sister chromatids during mitosis. How cohesin, an SMC complex that holds sister chromatids, affects TOP2α decatenation is unclear. To addres this, we developed a quadruple-trap optical tweezers assay to create DNA braids and study TOP2α decatenation at the single-molecule level in real-time. We show that TOP2α resolves both single and multiple braids but becomes inefficient at forces exceeding 28 pN. TOP2α is sensitive to DNA geometry, exhibiting a chiral preference for right-handed braid crossings, and requires loading directly at DNA crossovers to act. Pre-loading TOP2α onto individual DNA strands before braid formation, in the presence of ATP, prevents decatenation. Finally, we show that human cohesin, but not condensin I, binds stably to DNA braids and blocks TOP2α activity. Our study provides novel insights into the role of substrate accessibility in regulating TOP2α‘s activity and highlights cohesin as a barrier to decatenation.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © Crown 2025. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ |
Keywords: | DNA; Single-molecule biophysics |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Science (Sheffield) > School of Biosciences (Sheffield) |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 12 Aug 2025 09:31 |
Last Modified: | 12 Aug 2025 09:31 |
Status: | Published |
Publisher: | Springer Science and Business Media LLC |
Refereed: | Yes |
Identification Number: | 10.1038/s41467-025-62505-3 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:230277 |