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Phospholipid scramblase 1 interacts with influenza A virus NP, impairing its nuclear import and thereby suppressing virus replication
Transcription and replication of the influenza A virus (IAV) genome occur in the nucleus of infected cells and are carried out by the viral ribonucleoprotein complex (vRNP). As a major component of the vRNP complex, the viral nucleoprotein (NP) mediates the nuclear import of the vRNP complex via its...
Autores principales: | , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5792031/ https://www.ncbi.nlm.nih.gov/pubmed/29352288 http://dx.doi.org/10.1371/journal.ppat.1006851 |
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author | Luo, Weiyu Zhang, Jie Liang, Libin Wang, Guangwen Li, Qibing Zhu, Pengyang Zhou, Yuan Li, Junping Zhao, Yuhui Sun, Nan Huang, Shanyu Zhou, Chenchen Chang, Yu Cui, Pengfei Chen, Pucheng Jiang, Yongping Deng, Guohua Bu, Zhigao Li, Chengjun Jiang, Li Chen, Hualan |
author_facet | Luo, Weiyu Zhang, Jie Liang, Libin Wang, Guangwen Li, Qibing Zhu, Pengyang Zhou, Yuan Li, Junping Zhao, Yuhui Sun, Nan Huang, Shanyu Zhou, Chenchen Chang, Yu Cui, Pengfei Chen, Pucheng Jiang, Yongping Deng, Guohua Bu, Zhigao Li, Chengjun Jiang, Li Chen, Hualan |
author_sort | Luo, Weiyu |
collection | PubMed |
description | Transcription and replication of the influenza A virus (IAV) genome occur in the nucleus of infected cells and are carried out by the viral ribonucleoprotein complex (vRNP). As a major component of the vRNP complex, the viral nucleoprotein (NP) mediates the nuclear import of the vRNP complex via its nuclear localization signals (NLSs). Clearly, an effective way for the host to antagonize IAV infection would be by targeting vRNP nuclear import. Here, we identified phospholipid scramblase 1 (PLSCR1) as a binding partner of NP by using a yeast two-hybrid (Y2H) screen. The interaction between NP and PLSCR1 in mammalian cells was demonstrated by using co-immunoprecipitation and pull-down assays. We found that the stable overexpression of PLSCR1 suppressed the nuclear import of NP, hindered the virus life cycle, and significantly inhibited the replication of various influenza subtypes. In contrast, siRNA knockdown or CRISPR/Cas9 knockout of PLSCR1 increased virus propagation. Further analysis indicated that the inhibitory effect of PLSCR1 on the nuclear import of NP was not caused by affecting the phosphorylation status of NP or by stimulating the interferon (IFN) pathways. Instead, PLSCR1 was found to form a trimeric complex with NP and members of the importin α family, which inhibited the incorporation of importin β, a key mediator of the classical nuclear import pathway, into the complex, thus impairing the nuclear import of NP and suppressing virus replication. Our results demonstrate that PLSCR1 negatively regulates virus replication by interacting with NP in the cytoplasm and preventing its nuclear import. |
format | Online Article Text |
id | pubmed-5792031 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-57920312018-02-09 Phospholipid scramblase 1 interacts with influenza A virus NP, impairing its nuclear import and thereby suppressing virus replication Luo, Weiyu Zhang, Jie Liang, Libin Wang, Guangwen Li, Qibing Zhu, Pengyang Zhou, Yuan Li, Junping Zhao, Yuhui Sun, Nan Huang, Shanyu Zhou, Chenchen Chang, Yu Cui, Pengfei Chen, Pucheng Jiang, Yongping Deng, Guohua Bu, Zhigao Li, Chengjun Jiang, Li Chen, Hualan PLoS Pathog Research Article Transcription and replication of the influenza A virus (IAV) genome occur in the nucleus of infected cells and are carried out by the viral ribonucleoprotein complex (vRNP). As a major component of the vRNP complex, the viral nucleoprotein (NP) mediates the nuclear import of the vRNP complex via its nuclear localization signals (NLSs). Clearly, an effective way for the host to antagonize IAV infection would be by targeting vRNP nuclear import. Here, we identified phospholipid scramblase 1 (PLSCR1) as a binding partner of NP by using a yeast two-hybrid (Y2H) screen. The interaction between NP and PLSCR1 in mammalian cells was demonstrated by using co-immunoprecipitation and pull-down assays. We found that the stable overexpression of PLSCR1 suppressed the nuclear import of NP, hindered the virus life cycle, and significantly inhibited the replication of various influenza subtypes. In contrast, siRNA knockdown or CRISPR/Cas9 knockout of PLSCR1 increased virus propagation. Further analysis indicated that the inhibitory effect of PLSCR1 on the nuclear import of NP was not caused by affecting the phosphorylation status of NP or by stimulating the interferon (IFN) pathways. Instead, PLSCR1 was found to form a trimeric complex with NP and members of the importin α family, which inhibited the incorporation of importin β, a key mediator of the classical nuclear import pathway, into the complex, thus impairing the nuclear import of NP and suppressing virus replication. Our results demonstrate that PLSCR1 negatively regulates virus replication by interacting with NP in the cytoplasm and preventing its nuclear import. Public Library of Science 2018-01-19 /pmc/articles/PMC5792031/ /pubmed/29352288 http://dx.doi.org/10.1371/journal.ppat.1006851 Text en © 2018 Luo et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Luo, Weiyu Zhang, Jie Liang, Libin Wang, Guangwen Li, Qibing Zhu, Pengyang Zhou, Yuan Li, Junping Zhao, Yuhui Sun, Nan Huang, Shanyu Zhou, Chenchen Chang, Yu Cui, Pengfei Chen, Pucheng Jiang, Yongping Deng, Guohua Bu, Zhigao Li, Chengjun Jiang, Li Chen, Hualan Phospholipid scramblase 1 interacts with influenza A virus NP, impairing its nuclear import and thereby suppressing virus replication |
title | Phospholipid scramblase 1 interacts with influenza A virus NP, impairing its nuclear import and thereby suppressing virus replication |
title_full | Phospholipid scramblase 1 interacts with influenza A virus NP, impairing its nuclear import and thereby suppressing virus replication |
title_fullStr | Phospholipid scramblase 1 interacts with influenza A virus NP, impairing its nuclear import and thereby suppressing virus replication |
title_full_unstemmed | Phospholipid scramblase 1 interacts with influenza A virus NP, impairing its nuclear import and thereby suppressing virus replication |
title_short | Phospholipid scramblase 1 interacts with influenza A virus NP, impairing its nuclear import and thereby suppressing virus replication |
title_sort | phospholipid scramblase 1 interacts with influenza a virus np, impairing its nuclear import and thereby suppressing virus replication |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5792031/ https://www.ncbi.nlm.nih.gov/pubmed/29352288 http://dx.doi.org/10.1371/journal.ppat.1006851 |
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