The NS1 Protein from Influenza Virus Stimulates Translation Initiation by Enhancing Ribosome Recruitment to mRNAs

Panthu, Baptiste; Terrier, B.; Carron, Coralie; Traversier, Aurélien; Corbin, Antoine; Balvay, Laurent; Lina, Bruno; Rosa‐Calatrava, Manuel; Ohlmann, Théophile

doi:10.1016/j.jmb.2017.04.007

Cited by 26 publications

(39 citation statements)

References 57 publications

(70 reference statements)

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“…In addition to host shutoff, there is evidence that efficient translation of viral mRNAs depends on short, conserved cis-acting sequences in viral 5’ untranslated regions (UTRs), downstream of the host-derived leader sequences (6). It is not known precisely how these 5’-UTR sequences assist translation, but recent work has shown that when bound to viral mRNAs, the NS1 protein plays an important role in ribosome recruitment (7). By contrast, recent studies employing RNA-sequencing, ribosomal foot-printing and single-molecule fluorescence in-situ hybridization have suggested that host shutoff is mainly achieved by reduction in cellular mRNA levels, and that IAV mRNAs are not preferentially translated (8).…”

Section: Introductionmentioning

confidence: 99%

Stress granule-inducing eukaryotic translation initiation factor 4A inhibitors block influenza A virus replication

Slaine

Kleer

Smith

et al. 2017

Preprint

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Eukaryotic translation initiation factor 4A (eIF4A) is a helicase that facilitates assembly of the translation preinitiation complex by unwinding structured mRNA 5’ untranslated regions. Pateamine A (PatA) and silvestrol are natural products that disrupt eIF4A function and arrest translation, thereby triggering the formation of cytoplasmic aggregates of stalled preinitiation complexes known as stress granules (SGs). Here we examined the effects of eIF4A inhibition by PatA and silvestrol on influenza A virus (IAV) protein synthesis and replication in cell culture. Treatment of infected cells with either PatA or silvestrol at early times post-infection results in SG formation, arrest of viral protein synthesis and failure to replicate the viral genome. PatA, which irreversibly binds to eIF4A, sustained long-term blockade of IAV replication following drug withdrawal, and inhibited IAV replication at concentrations that had minimal cytotoxicity. By contrast, the antiviral effects of silvestrol were fully reversible; drug withdrawal caused rapid SG dissolution and resumption of viral protein synthesis. IAV inhibition by silvestrol was invariably associated with cytotoxicity. PatA blocked replication of genetically divergent IAV strains, suggesting common dependence on host eIF4A activity. This study demonstrates the feasibility of targeting core host protein synthesis machinery to prevent viral replication.IMPORTANCEInfluenza A virus (IAV) relies on cellular protein synthesis to decode viral messenger RNAs. Pateamine A and silvestrol are natural products that inactivate an essential protein synthesis protein known as eIF4A. Here we show that IAV is sensitive to these eIF4A inhibitor drugs. Treatment of infected cells with pateamine A or silvestrol prevented synthesis of viral proteins, viral genome replication and release of infectious virions. The irreversible eIF4A inhibitor pateamine A sustained long-term blockade of viral replication, whereas viral protein synthesis quickly resumed after silvestrol was removed from infected cells. Prolonged incubation of either infected or uninfected cells with these drugs induced the programmed cell death cascade called apoptosis. Our findings suggest that core components of the host protein synthesis machinery are viable targets for antiviral drug discovery. The most promising drug candidates should selectively block protein synthesis in infected cells without perturbing bystander uninfected cells.

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Section: Introductionmentioning

confidence: 99%

Stress granule-inducing eukaryotic translation initiation factor 4A inhibitors block influenza A virus replication

Slaine

Kleer

Smith

et al. 2017

Preprint

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“…NP protein forms viral ribonucleoprotein (RNP) complexes (nucleocapsids) with vRNA and, probably, acts as a negative translational regulator, closing the access of ribosomes to vRNA. The mechanism of positive regulation through the viral NS1 protein, which can enhance the translation of genomic RNA in infected cells, cannot be ruled out [19][20][21]. It is possible that cellular factors play a decisive role in the NSP expression and determine the tissue tropism of this protein synthesis in an infected organism [22].…”

Section: Discussionmentioning

confidence: 99%

Cellular immune response in infected mice to NSP protein encoded by the negative strand NS RNA of influenza A virus

Жирнов

Конакова

Anhlan

et al. 2019

Microbiology Independent Research Journal (MIR Journal)

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Influenza A virus belongs to a family of enveloped viruses with an RNA genome of negative polarity consisting of 8 RNA segments. The transcription of this RNA genome results in the synthesis of positive-sense mRNAs that translate up to 16 unique viral proteins with the help of splicing and translational shift mechanisms. The 8th NS segment encodes the NS1 protein (27 kDa), which is an active interferon antagonist, and the nuclear export protein NEP (14 kDa) through the standard negative polarity pathway. In addition, an alternative open reading frame for the synthesis of a third viral protein (NSP, negative-strand protein) by means of a direct translation of genome polarity RNA (the so-called positive polarity genome strategy) was identified in the NS segment. Since it is unknown as to whether the NSP protein can be synthesized in the infected organism post viral infection, the generation of spleen leucocytes specific to this protein was studied in mice after two sequential infections with influenza A viruses of H1N1 and H3N2 subtypes. It was found that leucocyte clones specifically recognizing a peptide domain in the central region of the NSP protein (amino acid positions 82-119) were generated in mice infected with influenza A viruses. In silico prediction has shown strong major histocompatibility complex-1 (MHC-I) and MHC-II specific epitopes in this central domain of the NSP. Comparative analysis of the influenza H3N2 viruses circulating in humans during 1968-2018 has shown high NSP variability, which was similar to that shown for the hemagglutinin (HA) and neuraminidase (NA) proteins. The highest variability was found to be in the N-and C-terminal parts of the NSP. These observations suggest that synthesis of the NSP protein occurs in infected animals and further support a bipolar (ambisense) strategy of the RNA genome of human influenza A virus.

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“…pCDNA-d2EGFP vector was generated by inserting the d2EGFP ORF into pCDNA3.1 (Life Technologies). pCDNA HIV-1 5´-UTR and pCDNA β-globin 5´-UTR were previously described (39). The dl HIV-1 IRES vector was previously described (40).…”

Section: Methodsmentioning

confidence: 99%

A Rev-CBP80-eIF4AI complex drives Gag synthesis from the HIV-1 unspliced mRNA

Toro-Ascuy

Rojas-Araya

García-de-Gracia

et al. 2018

Preprint

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33Gag synthesis from the full-length unspliced mRNA is critical for the production of the 34 viral progeny during human immunodeficiency virus type-1 (HIV-1) replication. While 35 most spliced mRNAs follow the canonical gene expression pathway in which the 36 recruitment of the nuclear cap-binding complex (CBC) and the exon junction complex 37 (EJC) largely stimulates the rates of nuclear export and translation, the unspliced mRNA 38 relies on the viral protein Rev to reach the cytoplasm and recruit the host translational 39 machinery. Here, we confirm that Rev ensures high levels of Gag synthesis by driving 40 nuclear export and translation of the unspliced mRNA. These functions of Rev are 41 supported by the CBC subunit CBP80, which binds Rev and the unspliced mRNA in the 42 nucleus and the cytoplasm. We also demonstrate that Rev interacts with the DEAD-box 43 RNA helicase eIF4AI, which translocates to the nucleus and cooperates with Rev to 44 promote Gag synthesis. Interestingly, molecular docking analyses revealed the assembly of 45 a Rev-CBP80-eIF4AI complex that is organized around the Rev response element (RRE). 46 Together, our results provide further evidence towards the understanding of the molecular 47 mechanisms by which Rev drives Gag synthesis from the unspliced mRNA during HIV-1 48 replication. 49 50 51 52 53 54 55 56 57 58 59 60 61 62Human Immunodeficiency Virus type-1 (HIV-1) gene expression is a complex process that 64 leads to the synthesis of fifteen proteins from one single primary transcript (1,2). Once the 65 proviral DNA has been integrated into the host cell genome, the RNA polymerase II drives 66 the synthesis of a 9-kb, capped and polyadenylated pre-mRNA that undergoes alternative 67 splicing generating more than 100 different transcripts classified into three main 68 populations (3,4). The so-called 2-kb multiply spliced transcripts code for the key 69 regulatory proteins Tat and Rev and the accessory protein Nef and are the dominant viral 70 mRNA species at early stages of viral gene expression (1,2,5). Unlike cellular mRNAs or 71 the 2-kb transcripts, which are spliced to completion before they exit the nucleus, HIV-1 72 and other complex retroviruses produce an important fraction of viral transcripts that 73 remain incompletely spliced (2,6). These 4-kb transcripts are expressed during the 74 intermediate phase of gene expression and are used for the synthesis of the envelope 75 glycoprotein (Env) and the accessory proteins Vif, Vpr and Vpu (2,6). Finally, the full-76 length 9-kb pre-mRNA in its unspliced form also reaches the cytoplasm to be used as an 77 mRNA template during the late stages of viral gene expression for the synthesis of the 78 major structural proteins Gag and Gag-Pol (1,2,6). 79Gene expression in eukaryotic cells occurs through the intricate connection of different 80 processes including transcription, splicing, nuclear export, translation and mRNA decay 81 and is regulated by the specific recruitment of nuclear proteins that together form the 82 messenger ribonuc...

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The NS1 Protein from Influenza Virus Stimulates Translation Initiation by Enhancing Ribosome Recruitment to mRNAs

Cited by 26 publications

References 57 publications

Stress granule-inducing eukaryotic translation initiation factor 4A inhibitors block influenza A virus replication

Stress granule-inducing eukaryotic translation initiation factor 4A inhibitors block influenza A virus replication

Cellular immune response in infected mice to NSP protein encoded by the negative strand NS RNA of influenza A virus

A Rev-CBP80-eIF4AI complex drives Gag synthesis from the HIV-1 unspliced mRNA

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