Chemical intervention of influenza virus mRNA nuclear export.
Influenza A viruses are human pathogens with limited therapeutic options. Therefore, it is crucial to devise strategies for the identification of new classes of antiviral medications. The influenza A virus genome is constituted of 8 RNA segments. Two of these viral RNAs are transcribed into mRNAs th...
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doaj-773ab3005ec64c18aba2b674724f2b622021-04-21T17:14:30ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742020-04-01164e100840710.1371/journal.ppat.1008407Chemical intervention of influenza virus mRNA nuclear export.Matthew EsparzaAmir MorHanspeter NiederstrasserKris WhiteAlexander WhiteKe ZhangShengyan GaoJuan WangJue LiangSei ShoRamanavelan SakthivelAdwait A SatheChao XingRaquel Muñoz-MorenoJerry W ShayAdolfo García-SastreJoseph ReadyBruce PosnerBeatriz M A FontouraInfluenza A viruses are human pathogens with limited therapeutic options. Therefore, it is crucial to devise strategies for the identification of new classes of antiviral medications. The influenza A virus genome is constituted of 8 RNA segments. Two of these viral RNAs are transcribed into mRNAs that are alternatively spliced. The M1 mRNA encodes the M1 protein but is also alternatively spliced to yield the M2 mRNA during infection. M1 to M2 mRNA splicing occurs at nuclear speckles, and M1 and M2 mRNAs are exported to the cytoplasm for translation. M1 and M2 proteins are critical for viral trafficking, assembly, and budding. Here we show that gene knockout of the cellular protein NS1-BP, a constituent of the M mRNA speckle-export pathway and a binding partner of the virulence factor NS1 protein, inhibits M mRNA nuclear export without altering bulk cellular mRNA export, providing an avenue to preferentially target influenza virus. We performed a high-content, image-based chemical screen using single-molecule RNA-FISH to label viral M mRNAs followed by multistep quantitative approaches to assess cellular mRNA and cell toxicity. We identified inhibitors of viral mRNA biogenesis and nuclear export that exhibited no significant activity towards bulk cellular mRNA at non-cytotoxic concentrations. Among the hits is a small molecule that preferentially inhibits nuclear export of a subset of viral and cellular mRNAs without altering bulk cellular mRNA export. These findings underscore specific nuclear export requirements for viral mRNAs and phenocopy down-regulation of the mRNA export factor UAP56. This RNA export inhibitor impaired replication of diverse influenza A virus strains at non-toxic concentrations. Thus, this screening strategy yielded compounds that alone or in combination may serve as leads to new ways of treating influenza virus infection and are novel tools for studying viral RNA trafficking in the nucleus.https://doi.org/10.1371/journal.ppat.1008407 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Matthew Esparza Amir Mor Hanspeter Niederstrasser Kris White Alexander White Ke Zhang Shengyan Gao Juan Wang Jue Liang Sei Sho Ramanavelan Sakthivel Adwait A Sathe Chao Xing Raquel Muñoz-Moreno Jerry W Shay Adolfo García-Sastre Joseph Ready Bruce Posner Beatriz M A Fontoura |
spellingShingle |
Matthew Esparza Amir Mor Hanspeter Niederstrasser Kris White Alexander White Ke Zhang Shengyan Gao Juan Wang Jue Liang Sei Sho Ramanavelan Sakthivel Adwait A Sathe Chao Xing Raquel Muñoz-Moreno Jerry W Shay Adolfo García-Sastre Joseph Ready Bruce Posner Beatriz M A Fontoura Chemical intervention of influenza virus mRNA nuclear export. PLoS Pathogens |
author_facet |
Matthew Esparza Amir Mor Hanspeter Niederstrasser Kris White Alexander White Ke Zhang Shengyan Gao Juan Wang Jue Liang Sei Sho Ramanavelan Sakthivel Adwait A Sathe Chao Xing Raquel Muñoz-Moreno Jerry W Shay Adolfo García-Sastre Joseph Ready Bruce Posner Beatriz M A Fontoura |
author_sort |
Matthew Esparza |
title |
Chemical intervention of influenza virus mRNA nuclear export. |
title_short |
Chemical intervention of influenza virus mRNA nuclear export. |
title_full |
Chemical intervention of influenza virus mRNA nuclear export. |
title_fullStr |
Chemical intervention of influenza virus mRNA nuclear export. |
title_full_unstemmed |
Chemical intervention of influenza virus mRNA nuclear export. |
title_sort |
chemical intervention of influenza virus mrna nuclear export. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS Pathogens |
issn |
1553-7366 1553-7374 |
publishDate |
2020-04-01 |
description |
Influenza A viruses are human pathogens with limited therapeutic options. Therefore, it is crucial to devise strategies for the identification of new classes of antiviral medications. The influenza A virus genome is constituted of 8 RNA segments. Two of these viral RNAs are transcribed into mRNAs that are alternatively spliced. The M1 mRNA encodes the M1 protein but is also alternatively spliced to yield the M2 mRNA during infection. M1 to M2 mRNA splicing occurs at nuclear speckles, and M1 and M2 mRNAs are exported to the cytoplasm for translation. M1 and M2 proteins are critical for viral trafficking, assembly, and budding. Here we show that gene knockout of the cellular protein NS1-BP, a constituent of the M mRNA speckle-export pathway and a binding partner of the virulence factor NS1 protein, inhibits M mRNA nuclear export without altering bulk cellular mRNA export, providing an avenue to preferentially target influenza virus. We performed a high-content, image-based chemical screen using single-molecule RNA-FISH to label viral M mRNAs followed by multistep quantitative approaches to assess cellular mRNA and cell toxicity. We identified inhibitors of viral mRNA biogenesis and nuclear export that exhibited no significant activity towards bulk cellular mRNA at non-cytotoxic concentrations. Among the hits is a small molecule that preferentially inhibits nuclear export of a subset of viral and cellular mRNAs without altering bulk cellular mRNA export. These findings underscore specific nuclear export requirements for viral mRNAs and phenocopy down-regulation of the mRNA export factor UAP56. This RNA export inhibitor impaired replication of diverse influenza A virus strains at non-toxic concentrations. Thus, this screening strategy yielded compounds that alone or in combination may serve as leads to new ways of treating influenza virus infection and are novel tools for studying viral RNA trafficking in the nucleus. |
url |
https://doi.org/10.1371/journal.ppat.1008407 |
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