Challenging the Roles of NSP3 and Untranslated Regions in Rotavirus mRNA Translation

Gratia, Matthieu; Vende, Patrice; Charpilienne, Annie; Baron, Hilma Carolina; Laroche, Cécile; Sarot, Emeline; Pyronnet, Stéphane; Duarte, Márcia do Rocio; Poncet, Didier

doi:10.1371/journal.pone.0145998

Cited by 18 publications

(20 citation statements)

References 49 publications

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“…NSP3 also engages the scaffolding protein eIF4G, which, in turn, interacts with the cap binding protein eIF4E (41). In this way, rotavirus recapitulates the mRNA circularization that enables efficient translation (42)(43)(44)(45)(46). Like rotavirus mRNAs, reovirus mRNAs contain a conserved tetranucleotide sequence (CATC) at their 3= termini (7).…”

Section: Discussionmentioning

confidence: 99%

Nonstructural Protein σ1s Is Required for Optimal Reovirus Protein Expression

Phillips

Stuart

Simon

2018

J Virol

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Reovirus non-structural protein σ1s is required for the establishment of viremia and hematogenous viral dissemination. However, the function of the σ1s protein during the reovirus replication cycle is not known. In this study, we found that σ1s was required for efficient reovirus replication in SV40-immortalized endothelial cells (SVECs), mouse embryonic fibroblasts, human umbilical vein endothelial cells (HUVECs), and T84 human colonic epithelial cells. In each of these cell lines, wild type reovirus produced substantially higher viral titers than a σ1s-deficient mutant. The σ1s protein was not required for early events in the reovirus infection, as no difference in infectivity between the wild type and σ1s-null viruses was observed. However, wild type virus produced markedly higher viral protein levels than the σ1s-deficient strain. The disparity in viral replication did not result from differences in viral transcription or protein stability. We further found that the σ1s protein was dispensable for cell killing and induction of type-1 interferon responses. In the absence of σ1s, viral factory (VF) maturation was impaired, but sufficient to support low levels of reovirus replication. Together, our results indicate that σ1s is not absolutely essential for viral protein production, but rather potentiates reovirus protein expression to facilitate reovirus replication. Our findings suggest that σ1s enables hematogenous reovirus dissemination by promoting efficient viral protein synthesis, and thereby reovirus replication, in cells that are required for reovirus spread to the blood.Hematogenous dissemination is critical a step in the pathogenesis of many viruses. For reovirus, nonstructural protein σ1s is required for viral spread via the blood. However, the mechanism by σ1s promotes reovirus dissemination is unknown. Here, we identified σ1s as a viral mediator of reovirus protein expression. We found several cultured cell lines in which σ1s is required for efficient reovirus replication. In these cells, wild type virus produced substantially higher levels of viral protein than a σ1s-deficient mutant. The σ1s protein was not required for viral mRNA transcription or viral protein stability. Owing to reduced levels of viral protein synthesized in the absence of σ1s, maturation of viral factories was impaired and significantly fewer viral progeny were produced. Taken together, our findings indicate that σ1s is required for optimal reovirus protein production, and thereby viral replication, in cells required hematogenous reovirus dissemination.

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Section: Discussionmentioning

confidence: 99%

Nonstructural Protein σ1s Is Required for Optimal Reovirus Protein Expression

Phillips

Stuart

Simon

2018

J Virol

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“…In both constructs, the nucleotide in position +4, which is not optimal in gs11 (U instead of G), was not changed. In a recent work with gs4 Gratia et al reported that optimisation of both positions -3 and +4 is important for expression efficiency [34]. However, the strong difference in expression of the two constructs with start codons sharing the same nucleotide context (GUG AUG U), strongly suggests that the IM inhibitory activity on translation cannot be ascribed to a less effective start codon recognition.…”

Section: Discussionmentioning

confidence: 99%

An Inhibitory Motif on the 5’UTR of Several Rotavirus Genome Segments Affects Protein Expression and Reverse Genetics Strategies

et al. 2016

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Rotavirus genome consists of eleven segments of dsRNA, each encoding one single protein. Viral mRNAs contain an open reading frame (ORF) flanked by relatively short untranslated regions (UTRs), whose role in the viral cycle remains elusive. Here we investigated the role of 5’UTRs in T7 polymerase-driven cDNAs expression in uninfected cells. The 5’UTRs of eight genome segments (gs3, gs5-6, gs7-11) of the simian SA11 strain showed a strong inhibitory effect on the expression of viral proteins. Decreased protein expression was due to both compromised transcription and translation and was independent of the ORF and the 3’UTR sequences. Analysis of several mutants of the 21-nucleotide long 5’UTR of gs 11 defined an inhibitory motif (IM) represented by its primary sequence rather than its secondary structure. IM was mapped to the 5’ terminal 6-nucleotide long pyrimidine-rich tract 5’-GGY(U/A)UY-3’. The 5’ terminal position within the mRNA was shown to be essentially required, as inhibitory activity was lost when IM was moved to an internal position. We identified two mutations (insertion of a G upstream the 5’UTR and the U to A mutation of the fifth nucleotide of IM) that render IM non-functional and increase the transcription and translation rate to levels that could considerably improve the efficiency of virus helper-free reverse genetics strategies.

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“…Second, NSP3 inactivates eIF2 by stimulating its phosphorylation. [46] Efficient translation of rotavirus mRNA, which lacks the 3' poly(A) tail, does not require either of these factors. [47] NSP4 is a viral enterotoxin that induces diarrhoea and was the first viral enterotoxin discovered.…”

Section: Proteinsmentioning

confidence: 99%

Rotavirus

Beards¹

2017

Wiki J Med

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Rotavirus is the most common cause of diarrhoeal disease among infants and young children. [1] It is a genus of double-stranded RNA viruses in the family Reoviridae. Nearly every child in the world is infected with rotavirus at least once by the age of five. [2] Immunity develops with each infection, so subsequent infections are less severe; adults are rarely affected. [3] There are eight species of this virus, referred to as A, B, C, D, E, F, G and H. Rotavirus A, the most common species, causes more than 90% of rotavirus infections in humans.The virus is transmitted by the faecal-oral route. It infects and damages the cells that line the small intestine and causes gastroenteritis (which is often called "stomach flu" despite having no relation to influenza). Although rotavirus was discovered in 1973 by Ruth Bishop and her colleagues by electron micrograph images [4] and accounts for approximately one third of hospitalisations for severe diarrhoea in infants and children, [5] its importance has historically been underestimated within the public health community, particularly in developing countries. [6] In addition to its impact on human health, rotavirus also infects animals, and is a pathogen of livestock. [7] Rotavirus is usually an easily managed disease of childhood, but in 2013, rotavirus caused 37 percent of deaths of children from diarrhoea and 215,000 deaths worldwide, [8] and almost two million more become severely ill. [6] Most of these deaths occurred in developing countries. [9] In the United States, before initiation of the rotavirus vaccination programme, rotavirus caused about 2.7 million cases of severe gastroenteritis in children, almost 60,000 hospitalisations, and around 37 deaths each year. [10] Following rotavirus vaccine introduction in the United States, hospitalisation rates have fallen significantly. [11][12] Public health campaigns to combat rotavirus focus on providing oral rehydration therapy for infected children and vaccination to prevent the disease. [13] The incidence and severity of rotavirus infections has declined significantly in countries that have added rotavirus vaccine to their routine childhood immunisation policies. [14][15] [16] Virology Types of rotavirusThere are eight species of rotavirus, referred to as groups A, B, C, D, E, F, G, and H. [17] Humans are primarily infected by species A, B and C, most commonly by species A. A-E species cause disease in other animals, [18] species E and H in pigs, and D, F and G in birds.[19] [20] Within rotavirus A there are different strains, called serotypes.[21] As with influenza virus, a dual classification system is used based on two proteins on the surface of the virus. The glycoprotein VP7 defines the G serotypes and the protease-sensitive protein VP4 defines P serotypes.[22] Because the two genes that determine G-types and P-types can be passed on separately to progeny viruses, different combinations are found.[22] A whole genome genotyping system has been established for group A rotaviruses, which has been used to ...

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Challenging the Roles of NSP3 and Untranslated Regions in Rotavirus mRNA Translation

Cited by 18 publications

References 49 publications

Nonstructural Protein σ1s Is Required for Optimal Reovirus Protein Expression

Nonstructural Protein σ1s Is Required for Optimal Reovirus Protein Expression

An Inhibitory Motif on the 5’UTR of Several Rotavirus Genome Segments Affects Protein Expression and Reverse Genetics Strategies

Rotavirus

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