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

Author:

Hurdiss Daniel L.ORCID,El Kazzi Priscila,Bauer Lisa,Papageorgiou Nicolas,Ferron François P.,Donselaar Tim,van Vliet Arno L.W.,Canard BrunoORCID,Decroly EtienneORCID,Brancale AndreaORCID,Zeev-Ben-Mordehai Tzviya,Förster Friedrich,van Kuppeveld Frank J.M,Coutard BrunoORCID

Abstract

AbstractThe enterovirus genus encompasses many clinically important human pathogens such as poliovirus, coxsackieviruses, echoviruses, numbered enteroviruses and rhinoviruses. These viruses are the etiological agents of several human diseases, including hand-foot-and-mouth disease, neonatal sepsis, encephalitis, meningitis, paralysis and respiratory infections. There is an unmet need for antivirals to treat these diseases. The non-structural protein 2C is a AAA+ helicase and plays a key role in viral replication. As such, it is an attractive target for antiviral drug development. Several repurposing screens with FDA-approved drugs have identified 2C-targeting compounds such as fluoxetine and dibucaine, but the molecular basis of 2C inhibition has remained enigmatic. Here we present the 1.5 Å resolution crystal structure of the soluble fragment of coxsackievirus B3 2C protein in complex with (S)-fluoxetine (SFX), which reveals a conserved, hydrophobic drug-binding pocket which is distal to the ATP binding site. To decipher the molecular mechanism of inhibition by fluoxetine and other 2C-targeting compounds, we engineered a soluble, hexameric and ATPase competent 2C protein. Using this system, we show that SFX, dibucaine, HBB and guanidine hydrochloride inhibit 2C ATPase activity in a dose-dependent manner. Moreover, using cryo-EM analysis, we demonstrate that SFX and dibucaine lock 2C in a defined hexameric state, rationalizing their mode of inhibition and allowing us to generate the first reconstruction of the oligomeric complex. Taken together, these results provide important structural and mechanistic insights into 2C inhibition and provide a robust engineering strategy which can be used for structural, functional and drug-screening analysis of 2C proteins from current or future enteroviruses.

Publisher

Cold Spring Harbor Laboratory

Reference60 articles.

1. Tapparel, C. , Siegrist, F. , Petty, T. J. & Kaiser, L. Picornavirus and enterovirus diversity with associated human diseases. Infection, Genetics and Evolution vol. 14 (2013).

2. Thibaut, H. J. et al. Toward antiviral therapy/prophylaxis for rhinovirus-induced exacerbations of chronic obstructive pulmonary disease: Challenges, opportunities, and strategies. Rev. Med. Virol. 26, (2016).

3. Chapman, N. M. & Kim, K. S. Persistent coxsackievirus infection: Enterovirus persistence in chronic myocarditis and dilated cardiomyopathy. Current Topics in Microbiology and Immunology vol. 323 (2008).

4. Puenpa, J. , Wanlapakorn, N. , Vongpunsawad, S. & Poovorawan, Y. The History of Enterovirus A71 Outbreaks and Molecular Epidemiology in the Asia-Pacific Region. Journal of Biomedical Science vol. 26 (2019).

5. Holm-Hansen, C. C. , Midgley, S. E. & Fischer, T. K. Global emergence of enterovirus D68: A systematic review. The Lancet Infectious Diseases vol. 16 (2016).

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1. Enterovirus A71 antivirals: Past, present, and future;Acta Pharmaceutica Sinica B;2021-08

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