Respiratory syncytial virus (RSV) enhances translation of virus-resembling AU-rich host transcripts

Abstract Background Viruses strongly rely on the host’s translational machinery to produce viral proteins required for replication. However, it is unknown how viruses that do not globally inhibit cap-dependent translation compete with abundant host transcripts for ribosomes. Viral infection often tr...

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Main Authors: Kyra Kerkhofs, Nicholas R. Guydosh, Mark A. Bayfield
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Virology Journal
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Online Access:https://doi.org/10.1186/s12985-025-02838-z
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author Kyra Kerkhofs
Nicholas R. Guydosh
Mark A. Bayfield
author_facet Kyra Kerkhofs
Nicholas R. Guydosh
Mark A. Bayfield
author_sort Kyra Kerkhofs
collection DOAJ
description Abstract Background Viruses strongly rely on the host’s translational machinery to produce viral proteins required for replication. However, it is unknown how viruses that do not globally inhibit cap-dependent translation compete with abundant host transcripts for ribosomes. Viral infection often triggers eukaryotic initiator factor 2α (eIF2α) phosphorylation, leading to global 5’-cap-dependent translation inhibition. Respiratory syncytial virus (RSV) encodes mRNAs mimicking 5’-cap structures of host mRNAs and thus inhibition of cap-dependent translation initiation would likely also reduce viral translation. Methods RSV-infected HEp-2 and A549 cells were analyzed to determine translation levels using western blotting, indirect immunofluorescent staining and polysome profiling. Transcriptome-wide translation efficiencies of virus-infected cells were compared against mock-infected cells using high-throughput sequencing of poly(A)-tail enriched total mRNA and transcripts associated with heavy polysomes. Results We confirmed that RSV limits widespread translation initiation inhibition and unexpectedly found that the fraction of ribosomes within polysomes increases during infection, indicating higher ribosome loading on mRNAs during infection. High-throughput sequencing revealed that virus-resembling, AU-rich host transcripts become more efficient at ribosome recruitment. Using a previously published dataset, we observe similar trends in another negative-sense single-stranded RNA virus, vesicular stomatitis virus (VSV). Conclusions These findings revealed that RSV changes the translational landscape by enhancing translation of virus-resembling AU-rich host transcripts rather than inhibiting host translation.
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spelling doaj-art-e179db629dac4a00bd38059bec598d632025-08-20T03:04:10ZengBMCVirology Journal1743-422X2025-07-0122111910.1186/s12985-025-02838-zRespiratory syncytial virus (RSV) enhances translation of virus-resembling AU-rich host transcriptsKyra Kerkhofs0Nicholas R. Guydosh1Mark A. Bayfield2Department of Biology, Faculty of Science, York UniversitySection On mRNA Regulation and Translation, Laboratory of Biochemistry & Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of HealthDepartment of Biology, Faculty of Science, York UniversityAbstract Background Viruses strongly rely on the host’s translational machinery to produce viral proteins required for replication. However, it is unknown how viruses that do not globally inhibit cap-dependent translation compete with abundant host transcripts for ribosomes. Viral infection often triggers eukaryotic initiator factor 2α (eIF2α) phosphorylation, leading to global 5’-cap-dependent translation inhibition. Respiratory syncytial virus (RSV) encodes mRNAs mimicking 5’-cap structures of host mRNAs and thus inhibition of cap-dependent translation initiation would likely also reduce viral translation. Methods RSV-infected HEp-2 and A549 cells were analyzed to determine translation levels using western blotting, indirect immunofluorescent staining and polysome profiling. Transcriptome-wide translation efficiencies of virus-infected cells were compared against mock-infected cells using high-throughput sequencing of poly(A)-tail enriched total mRNA and transcripts associated with heavy polysomes. Results We confirmed that RSV limits widespread translation initiation inhibition and unexpectedly found that the fraction of ribosomes within polysomes increases during infection, indicating higher ribosome loading on mRNAs during infection. High-throughput sequencing revealed that virus-resembling, AU-rich host transcripts become more efficient at ribosome recruitment. Using a previously published dataset, we observe similar trends in another negative-sense single-stranded RNA virus, vesicular stomatitis virus (VSV). Conclusions These findings revealed that RSV changes the translational landscape by enhancing translation of virus-resembling AU-rich host transcripts rather than inhibiting host translation.https://doi.org/10.1186/s12985-025-02838-zRSVVSVTranslation efficiencyPolysome profilingHigh-throughput sequencingAU-rich transcripts
spellingShingle Kyra Kerkhofs
Nicholas R. Guydosh
Mark A. Bayfield
Respiratory syncytial virus (RSV) enhances translation of virus-resembling AU-rich host transcripts
Virology Journal
RSV
VSV
Translation efficiency
Polysome profiling
High-throughput sequencing
AU-rich transcripts
title Respiratory syncytial virus (RSV) enhances translation of virus-resembling AU-rich host transcripts
title_full Respiratory syncytial virus (RSV) enhances translation of virus-resembling AU-rich host transcripts
title_fullStr Respiratory syncytial virus (RSV) enhances translation of virus-resembling AU-rich host transcripts
title_full_unstemmed Respiratory syncytial virus (RSV) enhances translation of virus-resembling AU-rich host transcripts
title_short Respiratory syncytial virus (RSV) enhances translation of virus-resembling AU-rich host transcripts
title_sort respiratory syncytial virus rsv enhances translation of virus resembling au rich host transcripts
topic RSV
VSV
Translation efficiency
Polysome profiling
High-throughput sequencing
AU-rich transcripts
url https://doi.org/10.1186/s12985-025-02838-z
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AT nicholasrguydosh respiratorysyncytialvirusrsvenhancestranslationofvirusresemblingaurichhosttranscripts
AT markabayfield respiratorysyncytialvirusrsvenhancestranslationofvirusresemblingaurichhosttranscripts