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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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2016-05-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fmicb.2016.00654/full |
| _version_ | 1828875764340097024 |
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| author | Sathya N. Thulasi Raman Yan eZhou |
| author_facet | Sathya N. Thulasi Raman Yan eZhou |
| author_sort | Sathya N. Thulasi Raman |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-13T08:05:55Z |
| format | Article |
| id | doaj.art-ad8ff1edc01b49f8944d7d2349176955 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-13T08:05:55Z |
| publishDate | 2016-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-ad8ff1edc01b49f8944d7d23491769552022-12-21T23:54:19ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2016-05-01710.3389/fmicb.2016.00654197910Networks of host factors that interact with NS1 protein of influenza A virusSathya N. Thulasi Raman0Yan eZhou1University of SaskatchewanUniversity of SaskatchewanPigs 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.http://journal.frontiersin.org/Journal/10.3389/fmicb.2016.00654/fullprotein interaction networksNS1 proteinSwine influenza virusStrep-tagswine respiratory epithelial cells |
| spellingShingle | Sathya N. Thulasi Raman Yan eZhou Networks of host factors that interact with NS1 protein of influenza A virus Frontiers in Microbiology protein interaction networks NS1 protein Swine influenza virus Strep-tag swine respiratory epithelial cells |
| 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 | protein interaction networks NS1 protein Swine influenza virus Strep-tag swine respiratory epithelial cells |
| url | http://journal.frontiersin.org/Journal/10.3389/fmicb.2016.00654/full |
| work_keys_str_mv | AT sathyanthulasiraman networksofhostfactorsthatinteractwithns1proteinofinfluenzaavirus AT yanezhou networksofhostfactorsthatinteractwithns1proteinofinfluenzaavirus |