A cytosolic disulfide bridge-supported dimerization is crucial for stability and cellular distribution of Coxsackievirus B3 protein 3A.
coxsackievirus B3
enterovirus
homodimerization
infection
protein 3A
Journal
The FEBS journal
ISSN: 1742-4658
Titre abrégé: FEBS J
Pays: England
ID NLM: 101229646
Informations de publication
Date de publication:
07 2022
07 2022
Historique:
revised:
20
12
2021
received:
05
10
2021
accepted:
20
01
2022
pubmed:
24
1
2022
medline:
9
7
2022
entrez:
23
1
2022
Statut:
ppublish
Résumé
RNA viruses in the Picornaviridae family express a large 250 kDa viral polyprotein that is processed by virus-encoded proteinases into mature functional proteins with specific functions for virus replication. One of these proteins is the highly conserved enteroviral transmembrane protein 3A that assists in reorganizing cellular membranes associated with the Golgi apparatus. Here, we studied the molecular properties of the Coxsackievirus B3 (CVB3) protein 3A with regard to its dimerization and its functional stability. By applying mutational analysis and biochemical characterization, we demonstrate that protein 3A forms DTT-sensitive disulfide-linked dimers via a conserved cytosolic cysteine residue at position 38 (Cys38). Homodimerization of CVB3 protein 3A via Cys38 leads to profound stabilization of the protein, whereas a C38A mutation promotes a rapid proteasome-dependent elimination of its monomeric form. The lysosomotropic agent chloroquine (CQ) exerted only minor stabilizing effects on the 3A monomer but resulted in enrichment of the homodimer. Our experimental data demonstrate that disulfide linkages via a highly conserved Cys-residue in enteroviral protein 3A have an important role in the dimerization of this viral protein, thereby preserving its stability and functional integrity.
Substances chimiques
Disulfides
0
Viral Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
3826-3838Informations de copyright
© 2022 The Authors. The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.
Références
Whitton JL, Cornell CT, Feuer R. Host and virus determinants of picornavirus pathogenesis and tropism. Nat Rev Microbiol. 2005;3:765-76.
Thibaut HJ, De Palma AM, Neyts J. Combating enterovirus replication: state-of-the-art on antiviral research. Biochem Pharmacol. 2012;83:185-92.
Baggen J, Thibaut HJ, Strating J, van Kuppeveld FJM. The life cycle of non-polio enteroviruses and how to target it. Nat Rev Microbiol. 2018;16:368-81.
Voss M, Braun V, Bredow C, Kloetzel P-M, Beling A. Coxsackievirus B3 exploits the ubiquitin-proteasome system to facilitate viral replication. Viruses. 2021;13:1360.
Wessels E, Notebaart RA, Duijsings D, Lanke K, Vergeer B, Melchers WJ, et al. Structure-function analysis of the coxsackievirus protein 3A: identification of residues important for dimerization, viral rna replication, and transport inhibition. J Biol Chem. 2006;281:28232-43.
Klima M, Chalupska D, Rozycki B, Humpolickova J, Rezabkova L, Silhan J, et al. Kobuviral non-structural 3A proteins act as molecular harnesses to hijack the host ACBD3 protein. Structure. 2017;25:219-30.
Horova V, Lyoo H, Różycki B, Chalupska D, Smola M, Humpolickova J, et al. Convergent evolution in the mechanisms of ACBD3 recruitment to picornavirus replication sites. PLoS Pathog. 2019;15:e1007962.
Smola M, Horova V, Boura E, Klima M. Structural basis for hijacking of the host ACBD3 protein by bovine and porcine enteroviruses and kobuviruses. Arch Virol. 2020;165:355-66.
Cornell CT, Kiosses WB, Harkins S, Whitton JL. Inhibition of protein trafficking by coxsackievirus B3: multiple viral proteins target a single organelle. J Virol. 2006;80:6637-47.
Strauss DM, Glustrom LW, Wuttke DS. Towards an understanding of the poliovirus replication complex: the solution structure of the soluble domain of the poliovirus 3A protein. J Mol Biol. 2003;330:225-34.
Berman HM, Battistuz T, Bhat TN, Bluhm WF, Bourne PE, Burkhardt K, et al. The protein data bank. Acta Crystallogr D Biol Crystallogr. 2002;58:899-907.
Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat Methods. 2012;9:671-675.