Cargando…
SERTA Domain Containing Protein 1 (SERTAD1) Interacts with Classical Swine Fever Virus Structural Glycoprotein E2, Which Is Involved in Virus Virulence in Swine
E2 is the major structural glycoprotein of the classical swine fever virus (CSFV). E2 has been shown to be involved in important virus functions such as replication and virulence in swine. Using the yeast two-hybrid system, we previously identified several host proteins specifically interacting with...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7232485/ https://www.ncbi.nlm.nih.gov/pubmed/32283651 http://dx.doi.org/10.3390/v12040421 |
_version_ | 1783535398299893760 |
---|---|
author | Vuono, Elizabeth A. Ramirez-Medina, Elizabeth Azzinaro, Paul Berggren, Keith A. Rai, Ayushi Pruitt, Sarah Silva, Ediane Velazquez-Salinas, Lauro Borca, Manuel V. Gladue, Douglas P. |
author_facet | Vuono, Elizabeth A. Ramirez-Medina, Elizabeth Azzinaro, Paul Berggren, Keith A. Rai, Ayushi Pruitt, Sarah Silva, Ediane Velazquez-Salinas, Lauro Borca, Manuel V. Gladue, Douglas P. |
author_sort | Vuono, Elizabeth A. |
collection | PubMed |
description | E2 is the major structural glycoprotein of the classical swine fever virus (CSFV). E2 has been shown to be involved in important virus functions such as replication and virulence in swine. Using the yeast two-hybrid system, we previously identified several host proteins specifically interacting with CSFV E2. Here, we analyze the protein interaction of E2 with SERTA domain containing protein 1 (SERTAD1), a factor involved in the stimulation of the transcriptional activities of different host genes. We have confirmed that the interaction between these two proteins occurs in CSFV-infected swine cells by using a proximity ligation assay and confocal microscopy. Amino acid residues in the CSFV E2 protein that are responsible for mediating the interaction with SERTAD1 were mapped by a yeast two-hybrid approach using a randomly mutated E2 library. Using that information, a recombinant CSFV mutant (E2ΔSERTAD1v) that harbors substitutions in those residues mediating the protein-interaction with SERTAD1 was developed and used to study the role of the E2-SERTAD1 interaction in viral replication and virulence in swine. CSFV E2ΔSERTAD1v, when compared to the parental BICv, showed a clearly decreased ability to replicate in the SK6 swine cell line and a more severe replication defect in primary swine macrophage cultures. Importantly, 80% of animals infected with E2ΔSERTAD1v survived infection, remaining clinically normal during the 21-day observational period. This result would indicate that the ability of CSFV E2 to bind host SERTAD1 protein during infection plays a critical role in virus virulence. |
format | Online Article Text |
id | pubmed-7232485 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-72324852020-05-22 SERTA Domain Containing Protein 1 (SERTAD1) Interacts with Classical Swine Fever Virus Structural Glycoprotein E2, Which Is Involved in Virus Virulence in Swine Vuono, Elizabeth A. Ramirez-Medina, Elizabeth Azzinaro, Paul Berggren, Keith A. Rai, Ayushi Pruitt, Sarah Silva, Ediane Velazquez-Salinas, Lauro Borca, Manuel V. Gladue, Douglas P. Viruses Article E2 is the major structural glycoprotein of the classical swine fever virus (CSFV). E2 has been shown to be involved in important virus functions such as replication and virulence in swine. Using the yeast two-hybrid system, we previously identified several host proteins specifically interacting with CSFV E2. Here, we analyze the protein interaction of E2 with SERTA domain containing protein 1 (SERTAD1), a factor involved in the stimulation of the transcriptional activities of different host genes. We have confirmed that the interaction between these two proteins occurs in CSFV-infected swine cells by using a proximity ligation assay and confocal microscopy. Amino acid residues in the CSFV E2 protein that are responsible for mediating the interaction with SERTAD1 were mapped by a yeast two-hybrid approach using a randomly mutated E2 library. Using that information, a recombinant CSFV mutant (E2ΔSERTAD1v) that harbors substitutions in those residues mediating the protein-interaction with SERTAD1 was developed and used to study the role of the E2-SERTAD1 interaction in viral replication and virulence in swine. CSFV E2ΔSERTAD1v, when compared to the parental BICv, showed a clearly decreased ability to replicate in the SK6 swine cell line and a more severe replication defect in primary swine macrophage cultures. Importantly, 80% of animals infected with E2ΔSERTAD1v survived infection, remaining clinically normal during the 21-day observational period. This result would indicate that the ability of CSFV E2 to bind host SERTAD1 protein during infection plays a critical role in virus virulence. MDPI 2020-04-09 /pmc/articles/PMC7232485/ /pubmed/32283651 http://dx.doi.org/10.3390/v12040421 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Vuono, Elizabeth A. Ramirez-Medina, Elizabeth Azzinaro, Paul Berggren, Keith A. Rai, Ayushi Pruitt, Sarah Silva, Ediane Velazquez-Salinas, Lauro Borca, Manuel V. Gladue, Douglas P. SERTA Domain Containing Protein 1 (SERTAD1) Interacts with Classical Swine Fever Virus Structural Glycoprotein E2, Which Is Involved in Virus Virulence in Swine |
title | SERTA Domain Containing Protein 1 (SERTAD1) Interacts with Classical Swine Fever Virus Structural Glycoprotein E2, Which Is Involved in Virus Virulence in Swine |
title_full | SERTA Domain Containing Protein 1 (SERTAD1) Interacts with Classical Swine Fever Virus Structural Glycoprotein E2, Which Is Involved in Virus Virulence in Swine |
title_fullStr | SERTA Domain Containing Protein 1 (SERTAD1) Interacts with Classical Swine Fever Virus Structural Glycoprotein E2, Which Is Involved in Virus Virulence in Swine |
title_full_unstemmed | SERTA Domain Containing Protein 1 (SERTAD1) Interacts with Classical Swine Fever Virus Structural Glycoprotein E2, Which Is Involved in Virus Virulence in Swine |
title_short | SERTA Domain Containing Protein 1 (SERTAD1) Interacts with Classical Swine Fever Virus Structural Glycoprotein E2, Which Is Involved in Virus Virulence in Swine |
title_sort | serta domain containing protein 1 (sertad1) interacts with classical swine fever virus structural glycoprotein e2, which is involved in virus virulence in swine |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7232485/ https://www.ncbi.nlm.nih.gov/pubmed/32283651 http://dx.doi.org/10.3390/v12040421 |
work_keys_str_mv | AT vuonoelizabetha sertadomaincontainingprotein1sertad1interactswithclassicalswinefevervirusstructuralglycoproteine2whichisinvolvedinvirusvirulenceinswine AT ramirezmedinaelizabeth sertadomaincontainingprotein1sertad1interactswithclassicalswinefevervirusstructuralglycoproteine2whichisinvolvedinvirusvirulenceinswine AT azzinaropaul sertadomaincontainingprotein1sertad1interactswithclassicalswinefevervirusstructuralglycoproteine2whichisinvolvedinvirusvirulenceinswine AT berggrenkeitha sertadomaincontainingprotein1sertad1interactswithclassicalswinefevervirusstructuralglycoproteine2whichisinvolvedinvirusvirulenceinswine AT raiayushi sertadomaincontainingprotein1sertad1interactswithclassicalswinefevervirusstructuralglycoproteine2whichisinvolvedinvirusvirulenceinswine AT pruittsarah sertadomaincontainingprotein1sertad1interactswithclassicalswinefevervirusstructuralglycoproteine2whichisinvolvedinvirusvirulenceinswine AT silvaediane sertadomaincontainingprotein1sertad1interactswithclassicalswinefevervirusstructuralglycoproteine2whichisinvolvedinvirusvirulenceinswine AT velazquezsalinaslauro sertadomaincontainingprotein1sertad1interactswithclassicalswinefevervirusstructuralglycoproteine2whichisinvolvedinvirusvirulenceinswine AT borcamanuelv sertadomaincontainingprotein1sertad1interactswithclassicalswinefevervirusstructuralglycoproteine2whichisinvolvedinvirusvirulenceinswine AT gladuedouglasp sertadomaincontainingprotein1sertad1interactswithclassicalswinefevervirusstructuralglycoproteine2whichisinvolvedinvirusvirulenceinswine |