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Networks of Host Factors that Interact with NS1 Protein of Influenza A Virus

Pigs are an important host of influenza A viruses due to their ability to generate reassortant viruses with pandemic potential. NS1 protein of influenza A viruses is a key virulence factor and a major antagonist of innate immune responses. It is also involved in enhancing viral mRNA translation and...

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Autores principales: Thulasi Raman, Sathya N., Zhou, Yan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4855030/
https://www.ncbi.nlm.nih.gov/pubmed/27199973
http://dx.doi.org/10.3389/fmicb.2016.00654
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author Thulasi Raman, Sathya N.
Zhou, Yan
author_facet Thulasi Raman, Sathya N.
Zhou, Yan
author_sort Thulasi Raman, Sathya N.
collection PubMed
description Pigs are an important host of influenza A viruses due to their ability to generate reassortant viruses with pandemic potential. NS1 protein of influenza A viruses is a key virulence factor and a major antagonist of innate immune responses. It is also involved in enhancing viral mRNA translation and regulation of virus replication. Being a protein with pleiotropic functions, NS1 has a variety of cellular interaction partners. Hence, studies on swine influenza viruses (SIV) and identification of swine influenza NS1-interacting host proteins is of great interest. Here, we constructed a recombinant SIV carrying a Strep-tag in the NS1 protein and infected primary swine respiratory epithelial cells (SRECs) with this virus. The Strep-tag sequence in the NS1 protein enabled us to purify intact, the NS1 protein and its interacting protein complex specifically. We identified cellular proteins present in the purified complex by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and generated a dataset of these proteins. 445 proteins were identified by LC-MS/MS and among them 192 proteins were selected by setting up a threshold based on MS parameters. The selected proteins were analyzed by bioinformatics and were categorized as belonging to different functional groups including translation, RNA processing, cytoskeleton, innate immunity, and apoptosis. Protein interaction networks were derived using these data and the NS1 interactions with some of the specific host factors were verified by immunoprecipitation. The novel proteins and the networks revealed in our study will be the potential candidates for targeted study of the molecular interaction of NS1 with host proteins, which will provide insights into the identification of new therapeutic targets to control influenza infection and disease pathogenesis.
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spelling pubmed-48550302016-05-19 Networks of Host Factors that Interact with NS1 Protein of Influenza A Virus Thulasi Raman, Sathya N. Zhou, Yan Front Microbiol Microbiology Pigs are an important host of influenza A viruses due to their ability to generate reassortant viruses with pandemic potential. NS1 protein of influenza A viruses is a key virulence factor and a major antagonist of innate immune responses. It is also involved in enhancing viral mRNA translation and regulation of virus replication. Being a protein with pleiotropic functions, NS1 has a variety of cellular interaction partners. Hence, studies on swine influenza viruses (SIV) and identification of swine influenza NS1-interacting host proteins is of great interest. Here, we constructed a recombinant SIV carrying a Strep-tag in the NS1 protein and infected primary swine respiratory epithelial cells (SRECs) with this virus. The Strep-tag sequence in the NS1 protein enabled us to purify intact, the NS1 protein and its interacting protein complex specifically. We identified cellular proteins present in the purified complex by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and generated a dataset of these proteins. 445 proteins were identified by LC-MS/MS and among them 192 proteins were selected by setting up a threshold based on MS parameters. The selected proteins were analyzed by bioinformatics and were categorized as belonging to different functional groups including translation, RNA processing, cytoskeleton, innate immunity, and apoptosis. Protein interaction networks were derived using these data and the NS1 interactions with some of the specific host factors were verified by immunoprecipitation. The novel proteins and the networks revealed in our study will be the potential candidates for targeted study of the molecular interaction of NS1 with host proteins, which will provide insights into the identification of new therapeutic targets to control influenza infection and disease pathogenesis. Frontiers Media S.A. 2016-05-04 /pmc/articles/PMC4855030/ /pubmed/27199973 http://dx.doi.org/10.3389/fmicb.2016.00654 Text en Copyright © 2016 Thulasi Raman and Zhou. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Thulasi Raman, Sathya N.
Zhou, Yan
Networks of Host Factors that Interact with NS1 Protein of Influenza A Virus
title Networks of Host Factors that Interact with NS1 Protein of Influenza A Virus
title_full Networks of Host Factors that Interact with NS1 Protein of Influenza A Virus
title_fullStr Networks of Host Factors that Interact with NS1 Protein of Influenza A Virus
title_full_unstemmed Networks of Host Factors that Interact with NS1 Protein of Influenza A Virus
title_short Networks of Host Factors that Interact with NS1 Protein of Influenza A Virus
title_sort networks of host factors that interact with ns1 protein of influenza a virus
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4855030/
https://www.ncbi.nlm.nih.gov/pubmed/27199973
http://dx.doi.org/10.3389/fmicb.2016.00654
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