Polyprotein processing and intermolecular interactions within the viral replication complex spatially and temporally control norovirus protease activity

Emmott, Edward ORCID: 0000-0002-3239-8178, de Rougemont, Alexis, Hosmillo, Myra, Lu, Jia, Fitzmaurice, Timothy, Haas, Juergen and Goodfellow, Ian
(2019) Polyprotein processing and intermolecular interactions within the viral replication complex spatially and temporally control norovirus protease activity. JOURNAL OF BIOLOGICAL CHEMISTRY, 294 (11). pp. 4259-4271.

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Norovirus infections are a major cause of acute viral gastroenteritis and a significant burden on global human health. A vital process for norovirus replication is the processing of the nonstructural polyprotein by a viral protease into the viral components required to form the viral replication complex. This cleavage occurs at different rates, resulting in the accumulation of stable precursor forms. Here, we characterized how precursor forms of the norovirus protease accumulate during infection. Using stable forms of the protease precursors, we demonstrated that all of them are proteolytically active <i>in vitro</i>, but that when expressed in cells, their activities are determined by both substrate and protease localization. Although all precursors could cleave a replication complex-associated substrate, only a subset of precursors lacking the NS4 protein were capable of efficiently cleaving a cytoplasmic substrate. By mapping the full range of protein-protein interactions among murine and human norovirus proteins with the LUMIER assay, we uncovered conserved interactions between replication complex members that modify the localization of a protease precursor subset. Finally, we demonstrate that fusion to the membrane-bound replication complex components permits efficient cleavage of a fused substrate when active polyprotein-derived protease is provided in <i>trans</i> These findings offer a model for how norovirus can regulate the timing of substrate cleavage throughout the replication cycle. Because the norovirus protease represents a key target in antiviral therapies, an improved understanding of its function and regulation, as well as identification of interactions among the other nonstructural proteins, offers new avenues for antiviral drug design.

Item Type: Article
Uncontrolled Keywords: protease, fluorescence resonance energy transfer (FRET), viral replication, plus-stranded RNA virus, RNA virus, Calicivirus, Norovirus, Precursor
Divisions: Faculty of Health and Life Sciences
Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology
Depositing User: Symplectic Admin
Date Deposited: 24 Apr 2023 07:25
Last Modified: 24 Apr 2023 07:25
DOI: 10.1074/jbc.RA118.006780
Open Access URL: https://www.jbc.org/article/S0021-9258(20)41831-9/...
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3169898