A Conserved Interaction between a C-Terminal Motif in Norovirus VPg and the HEAT-1 Domain of eIF4G Is Essential for Translation Initiation
dc.contributor.author
Leen, Eoin N.
dc.contributor.author
Sorgeloos, Frédéric
dc.contributor.author
Correia, Samantha
dc.contributor.author
Chaudhry, Yasmin
dc.contributor.author
Cannac, Fabien
dc.contributor.author
Pastore, Chiara
dc.contributor.author
Xu, Yingqi
dc.contributor.author
Graham, Stephen C.
dc.contributor.author
Matthews, Stephen J.
dc.contributor.author
Goodfellow, Ian G.
dc.contributor.author
Curry, Stephen
dc.date.accessioned
2018-11-06T17:57:10Z
dc.date.available
2017-06-12T01:30:43Z
dc.date.available
2018-11-06T17:57:10Z
dc.date.issued
2016-01-06
dc.identifier.issn
1553-7374
dc.identifier.issn
1553-7366
dc.identifier.other
10.1371/journal.ppat.1005379
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/113392
dc.identifier.doi
10.3929/ethz-b-000113392
dc.description.abstract
Translation initiation is a critical early step in the replication cycle of the positive-sense, single-stranded RNA genome of noroviruses, a major cause of gastroenteritis in humans. Norovirus RNA, which has neither a 5´ m7G cap nor an internal ribosome entry site (IRES), adopts an unusual mechanism to initiate protein synthesis that relies on interactions between the VPg protein covalently attached to the 5´-end of the viral RNA and eukaryotic initiation factors (eIFs) in the host cell. For murine norovirus (MNV) we previously showed that VPg binds to the middle fragment of eIF4G (4GM; residues 652–1132). Here we have used pull-down assays, fluorescence anisotropy, and isothermal titration calorimetry (ITC) to demonstrate that a stretch of ~20 amino acids at the C terminus of MNV VPg mediates direct and specific binding to the HEAT-1 domain within the 4GM fragment of eIF4G. Our analysis further reveals that the MNV C terminus binds to eIF4G HEAT-1 via a motif that is conserved in all known noroviruses. Fine mutagenic mapping suggests that the MNV VPg C terminus may interact with eIF4G in a helical conformation. NMR spectroscopy was used to define the VPg binding site on eIF4G HEAT-1, which was confirmed by mutagenesis and binding assays. We have found that this site is non-overlapping with the binding site for eIF4A on eIF4G HEAT-1 by demonstrating that norovirus VPg can form ternary VPg-eIF4G-eIF4A complexes. The functional significance of the VPg-eIF4G interaction was shown by the ability of fusion proteins containing the C-terminal peptide of MNV VPg to inhibit in vitro translation of norovirus RNA but not cap- or IRES-dependent translation. These observations define important structural details of a functional interaction between norovirus VPg and eIF4G and reveal a binding interface that might be exploited as a target for antiviral therapy.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
PLOS
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.title
A Conserved Interaction between a C-Terminal Motif in Norovirus VPg and the HEAT-1 Domain of eIF4G Is Essential for Translation Initiation
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 4.0 International
ethz.journal.title
PLoS Pathogens
ethz.journal.volume
12
en_US
ethz.journal.issue
1
en_US
ethz.journal.abbreviated
PLoS Pathog
ethz.pages.start
e1005379
en_US
ethz.size
34 p.
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.identifier.wos
ethz.identifier.scopus
ethz.publication.place
Lawrence, KS
ethz.publication.status
published
en_US
ethz.date.deposited
2017-06-12T01:34:23Z
ethz.source
ECIT
ethz.identifier.importid
imp59365426df97420238
ethz.ecitpid
pub:175083
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2017-07-13T12:48:40Z
ethz.rosetta.lastUpdated
2024-02-02T06:33:43Z
ethz.rosetta.versionExported
true
ethz.COinS
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