Elizabeth Michalczyk
Structural basis of chiral wrap and T-segment capture by Escherichia coli DNA gyrase
Michalczyk, Elizabeth; Pakosz-Stępień, Zuzanna; Liston, Jonathon D.; Gittins, Olivia; Pabis, Marta; Heddle, Jonathan G.; Ghilarov, Dmitry
Authors
Zuzanna Pakosz-Stepien zuzanna.pakosz-stepien@durham.ac.uk
Postdoctoral Research Associate
Dr Jonathon Liston jonathon.d.liston@durham.ac.uk
Postdoctoral Research Associate
Olivia Gittins olivia.v.gittins@durham.ac.uk
Postdoctoral Research Associate
Marta Pabis
Professor Jonathan Heddle jonathan.g.heddle@durham.ac.uk
Leverhulme International Professor
Dmitry Ghilarov
Abstract
Type II topoisomerase DNA gyrase transduces the energy of ATP hydrolysis into the negative supercoiling of DNA. The postulated catalytic mechanism involves stabilization of a chiral DNA loop followed by the passage of the T-segment through the temporarily cleaved G-segment resulting in sign inversion. The molecular basis for this is poorly understood as the chiral loop has never been directly observed. We have obtained high-resolution cryoEM structures of Escherichia coli gyrase with chirally wrapped 217 bp DNA with and without the fluoroquinolone moxifloxacin (MFX). Each structure constrains a positively supercoiled figure-of-eight DNA loop stabilized by a GyrA β-pinwheel domain which has the structure of a flat disc. By comparing the catalytic site of the native drug-free and MFX-bound gyrase structures both of which contain a single metal ion, we demonstrate that the enzyme is observed in a native precatalytic state. Our data imply that T-segment trapping is not dependent on the dimerization of the ATPase domains which appears to only be possible after strand passage has taken place.
Citation
Michalczyk, E., Pakosz-Stępień, Z., Liston, J. D., Gittins, O., Pabis, M., Heddle, J. G., & Ghilarov, D. (2024). Structural basis of chiral wrap and T-segment capture by Escherichia coli DNA gyrase. Proceedings of the National Academy of Sciences, 121(49), Article e2407398121. https://doi.org/10.1073/pnas.2407398121
Journal Article Type | Article |
---|---|
Acceptance Date | Oct 17, 2024 |
Online Publication Date | Nov 26, 2024 |
Publication Date | Dec 3, 2024 |
Deposit Date | Nov 27, 2024 |
Publicly Available Date | Nov 27, 2024 |
Journal | Proceedings of the National Academy of Sciences |
Print ISSN | 0027-8424 |
Electronic ISSN | 1091-6490 |
Publisher | National Academy of Sciences |
Peer Reviewed | Peer Reviewed |
Volume | 121 |
Issue | 49 |
Article Number | e2407398121 |
DOI | https://doi.org/10.1073/pnas.2407398121 |
Keywords | Models, Molecular, Escherichia coli - metabolism - enzymology - genetics, DNA Gyrase - metabolism - chemistry, Moxifloxacin - chemistry, Cryoelectron Microscopy, Catalytic Domain, Topoisomerase II Inhibitors - chemistry - pharmacology, antibiotics, DNA crossover, topoisomerase, DNA, Superhelical - metabolism - chemistry, molecular machine, Nucleic Acid Conformation, DNA-binding protein, DNA, Bacterial - metabolism, Fluoroquinolones - chemistry - pharmacology |
Public URL | https://durham-repository.worktribe.com/output/3107520 |
Files
Published Journal Article
(3.4 Mb)
PDF
Publisher Licence URL
http://creativecommons.org/licenses/by/4.0/
You might also like
A DNA Origami Bubble Blower for Liposome Production
(2024)
Journal Article
Reengineering of an Artificial Protein Cage for Efficient Packaging of Active Enzymes
(2024)
Journal Article
Downloadable Citations
About Durham Research Online (DRO)
Administrator e-mail: dro.admin@durham.ac.uk
This application uses the following open-source libraries:
SheetJS Community Edition
Apache License Version 2.0 (http://www.apache.org/licenses/)
PDF.js
Apache License Version 2.0 (http://www.apache.org/licenses/)
Font Awesome
SIL OFL 1.1 (http://scripts.sil.org/OFL)
MIT License (http://opensource.org/licenses/mit-license.html)
CC BY 3.0 ( http://creativecommons.org/licenses/by/3.0/)
Powered by Worktribe © 2025
Advanced Search