PXD074413-1

PXD074413 is an original dataset announced via ProteomeXchange.

Title	Hydrogen-deuterium exchange points to interactions at the helicase - topoisomerase interface in reverse gyrase that play a role in positive DNA supercoiling
Description	Reverse gyrase is a DNA topoisomerase that catalyzes positive DNA supercoiling in an ATP-dependent reaction. The enzyme consists of a helicase and a topoisomerase domain. The isolated helicase-like domain is a DNA-stimulated ATPase, and the isolated topoisomerase domain can relax supercoiled DNA. For positive supercoiling of DNA, the two domains need to cooperate, presumably through conformational changes. Here, we probed conformational changes of Thermotoga maritima reverse gyrase on DNA and nucleotide binding using hydrogen deuterium exchange (HDX). While ADPNP binding does not induce major conformational changes, binding of DNA leads to increased exposure of a region at the helicase-topoisomerase interface, encompassing R501 and E705, the side chains of which engage in electrostatic interactions. Removing the charges in an R501/E705A variant leads to a moderate increase in supercoiling activity, suggesting a slightly inhibitory role of this interactions. A R501/E705C variant, crosslinked to fix the two side chains in a disulphide bond, on the other hand, is inactive. Collectively these data show that rearrangements at the helicase and topoisomerase interface are important for the functional cooperation of the domains and for the enzymatic activity of reverse gyrase.
HostingRepository	PRIDE
AnnounceDate	2026-08-24
AnnouncementXML	Submission_2026-08-23_16:10:23.567.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Wieland Steinchen
SpeciesList	scientific name: Thermotoga maritima; NCBI TaxID: NEWT:2336;
ModificationList	No PTMs are included in the dataset
Instrument	Synapt MS

Revision	Datetime	Status	ChangeLog Entry
0	2026-02-12 15:43:29	ID requested
⏵ 1	2026-08-23 16:10:24	announced

10.1016/j.jbc.2026.113233;

Mhaindarkar VP, Collin F, Koldewey P, Klostermeier D, Hydrogen-deuterium exchange points to inter-domain interactions in reverse gyrase that modulate conformational changes in positive DNA supercoiling. J Biol Chem, 302(7):113233(2026) [pubmed]

ProteomeXchange project tag: Hydrogen Deuterium Exchange (HDX-MS)

submitter keyword: topoisomerase, helicase,reverse gyrase, conformational changes, positive DNA supercoiling

Dagmar Klostermeier
contact affiliation	University of Muenster, Institute for Physical Chemistry, Corrensstrasse 30, 48149 Muenster, Germany
contact email	dagmar.klostermeier@uni-muenster.de
lab head
Wieland Steinchen
contact affiliation	Philipps-University Marburg, Department of Chemistry & SYNMIKRO, Karl-von-Frisch-Straße 14, 35043 Marburg, GERMANY
contact email	wieland.steinchen@synmikro.uni-marburg.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/08/PXD074413

PRIDE project URI

• PRIDE
  1. PXD074413
     1. Label: PRIDE project
     2. Name: Hydrogen-deuterium exchange points to interactions at the helicase - topoisomerase interface in reverse gyrase that play a role in positive DNA supercoiling