The RNA pseudoknots in foot-and-mouth disease virus are dispensable for genome replication, but essential for the production of infectious virus.

Non-coding regions of viral RNA (vRNA) genomes are critically important in the regulation of gene expression. In particular, pseudoknot (PK) structures, which are present in a wide range of RNA molecules, have a variety of roles. The 5' untranslated region (5' UTR) of foot-and-mouth diseas...

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Main Authors: Joseph C Ward, Lidia Lasecka-Dykes, Chris Neil, Oluwapelumi O Adeyemi, Sarah Gold, Niall McLean-Pell, Caroline Wright, Morgan R Herod, David Kealy, Emma Warner, Terry Jackson, Donald P King, Tobias J Tuthill, David J Rowlands, Nicola J Stonehouse
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2022-06-01
Series:PLoS Pathogens
Online Access:https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1010589&type=printable
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author Joseph C Ward
Lidia Lasecka-Dykes
Chris Neil
Oluwapelumi O Adeyemi
Sarah Gold
Niall McLean-Pell
Caroline Wright
Morgan R Herod
David Kealy
Emma Warner
Terry Jackson
Donald P King
Tobias J Tuthill
David J Rowlands
Nicola J Stonehouse
author_facet Joseph C Ward
Lidia Lasecka-Dykes
Chris Neil
Oluwapelumi O Adeyemi
Sarah Gold
Niall McLean-Pell
Caroline Wright
Morgan R Herod
David Kealy
Emma Warner
Terry Jackson
Donald P King
Tobias J Tuthill
David J Rowlands
Nicola J Stonehouse
author_sort Joseph C Ward
collection DOAJ
description Non-coding regions of viral RNA (vRNA) genomes are critically important in the regulation of gene expression. In particular, pseudoknot (PK) structures, which are present in a wide range of RNA molecules, have a variety of roles. The 5' untranslated region (5' UTR) of foot-and-mouth disease virus (FMDV) vRNA is considerably longer than in other viruses from the picornavirus family and consists of a number of distinctive structural motifs that includes multiple (2, 3 or 4 depending on the virus strain) putative PKs linked in tandem. The role(s) of the PKs in the FMDV infection are not fully understood. Here, using bioinformatics, sub-genomic replicons and recombinant viruses we have investigated the structural conservation and importance of the PKs in the FMDV lifecycle. Our results show that despite the conservation of two or more PKs across all FMDVs, a replicon lacking PKs was replication competent, albeit at reduced levels. Furthermore, in competition experiments, GFP FMDV replicons with less than two (0 or 1) PK structures were outcompeted by a mCherry FMDV wt replicon that had 4 PKs, whereas GFP replicons with 2 or 4 PKs were not. This apparent replicative advantage offered by the additional PKs correlates with the maintenance of at least two PKs in the genomes of FMDV field isolates. Despite a replicon lacking any PKs retaining the ability to replicate, viruses completely lacking PK were not viable and at least one PK was essential for recovery of infections virus, suggesting a role for the PKs in virion assembly. Thus, our study points to roles for the PKs in both vRNA replication and virion assembly, thereby improving understanding the molecular biology of FMDV replication and the wider roles of PK in RNA functions.
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spelling doaj-art-8c98451b274449f58dae2fd9d06a9bcd2025-08-20T03:25:46ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742022-06-01186e101058910.1371/journal.ppat.1010589The RNA pseudoknots in foot-and-mouth disease virus are dispensable for genome replication, but essential for the production of infectious virus.Joseph C WardLidia Lasecka-DykesChris NeilOluwapelumi O AdeyemiSarah GoldNiall McLean-PellCaroline WrightMorgan R HerodDavid KealyEmma WarnerTerry JacksonDonald P KingTobias J TuthillDavid J RowlandsNicola J StonehouseNon-coding regions of viral RNA (vRNA) genomes are critically important in the regulation of gene expression. In particular, pseudoknot (PK) structures, which are present in a wide range of RNA molecules, have a variety of roles. The 5' untranslated region (5' UTR) of foot-and-mouth disease virus (FMDV) vRNA is considerably longer than in other viruses from the picornavirus family and consists of a number of distinctive structural motifs that includes multiple (2, 3 or 4 depending on the virus strain) putative PKs linked in tandem. The role(s) of the PKs in the FMDV infection are not fully understood. Here, using bioinformatics, sub-genomic replicons and recombinant viruses we have investigated the structural conservation and importance of the PKs in the FMDV lifecycle. Our results show that despite the conservation of two or more PKs across all FMDVs, a replicon lacking PKs was replication competent, albeit at reduced levels. Furthermore, in competition experiments, GFP FMDV replicons with less than two (0 or 1) PK structures were outcompeted by a mCherry FMDV wt replicon that had 4 PKs, whereas GFP replicons with 2 or 4 PKs were not. This apparent replicative advantage offered by the additional PKs correlates with the maintenance of at least two PKs in the genomes of FMDV field isolates. Despite a replicon lacking any PKs retaining the ability to replicate, viruses completely lacking PK were not viable and at least one PK was essential for recovery of infections virus, suggesting a role for the PKs in virion assembly. Thus, our study points to roles for the PKs in both vRNA replication and virion assembly, thereby improving understanding the molecular biology of FMDV replication and the wider roles of PK in RNA functions.https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1010589&type=printable
spellingShingle Joseph C Ward
Lidia Lasecka-Dykes
Chris Neil
Oluwapelumi O Adeyemi
Sarah Gold
Niall McLean-Pell
Caroline Wright
Morgan R Herod
David Kealy
Emma Warner
Terry Jackson
Donald P King
Tobias J Tuthill
David J Rowlands
Nicola J Stonehouse
The RNA pseudoknots in foot-and-mouth disease virus are dispensable for genome replication, but essential for the production of infectious virus.
PLoS Pathogens
title The RNA pseudoknots in foot-and-mouth disease virus are dispensable for genome replication, but essential for the production of infectious virus.
title_full The RNA pseudoknots in foot-and-mouth disease virus are dispensable for genome replication, but essential for the production of infectious virus.
title_fullStr The RNA pseudoknots in foot-and-mouth disease virus are dispensable for genome replication, but essential for the production of infectious virus.
title_full_unstemmed The RNA pseudoknots in foot-and-mouth disease virus are dispensable for genome replication, but essential for the production of infectious virus.
title_short The RNA pseudoknots in foot-and-mouth disease virus are dispensable for genome replication, but essential for the production of infectious virus.
title_sort rna pseudoknots in foot and mouth disease virus are dispensable for genome replication but essential for the production of infectious virus
url https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1010589&type=printable
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