Fluoxetine targets an allosteric site in the enterovirus 2C AAA+ ATPase and stabilizes a ring-shaped hexameric complex

Hurdiss, Daniel L., El Kazzi, Priscila, Bauer, Lisa, Papageorgiou, Nicolas, Ferron, François P., Donselaar, Tim, van Vliet, Arno L.W., Shamorkina, Tatiana M., Snijder, Joost, Canard, Bruno, Decroly, Etienne, Brancale, Andrea ORCID: https://orcid.org/0000-0002-9728-3419, Zeev-Ben-Mordehai, Tzviya, Förster, Friedrich, van Kuppeveld, Frank J.M. and Coutard, Bruno 2022. Fluoxetine targets an allosteric site in the enterovirus 2C AAA+ ATPase and stabilizes a ring-shaped hexameric complex. Science Advances 8 (1) , eabj7615. 10.1126/sciadv.abj7615

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Abstract

Enteroviruses are globally prevalent human pathogens responsible for many diseases. The nonstructural protein 2C is a AAA+ helicase and plays a key role in enterovirus replication. Drug repurposing screens identified 2C-targeting compounds such as fluoxetine and dibucaine, but how they inhibit 2C is unknown. Here, we present a crystal structure of the soluble and monomeric fragment of coxsackievirus B3 2C protein in complex with (S)-fluoxetine (SFX), revealing an allosteric binding site. To study the functional consequences of SFX binding, we engineered an adenosine triphosphatase (ATPase)–competent, hexameric 2C protein. Using this system, we show that SFX, dibucaine, HBB [2-(α-hydroxybenzyl)-benzimidazole], and guanidine hydrochloride inhibit 2C ATPase activity. Moreover, cryo–electron microscopy analysis demonstrated that SFX and dibucaine lock 2C in a defined hexameric state, rationalizing their mode of inhibition. Collectively, these results provide important insights into 2C inhibition and a robust engineering strategy for structural, functional, and drug-screening analysis of 2C proteins.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Pharmacy
Additional Information:	Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).
Publisher:	American Association for the Advancement of Science
ISSN:	2375-2548
Date of First Compliant Deposit:	26 January 2022
Date of Acceptance:	10 November 2021
Last Modified:	05 Jan 2024 05:51
URI:	https://orca.cardiff.ac.uk/id/eprint/146989

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