A β-Hairpin Motif in the Envelope Protein E2 Mediates Receptor Binding of Bovine Viral Diarrhea Virus
Pestivirus envelope protein E2 is crucial to virus infection and accomplishes virus-receptor interaction during entry. However, mapping of E2 residues mediating these interactions has remained unexplored. In this study, to investigate the structure-function relationship for a β-hairpin motif exposed to the solvent in the crystal structure of bovine viral diarrhea virus (BVDV) E2, we designed two amino acidic substitutions that result in a change of electrostatic potential. First, using wild type and mutant E2 expressed as soluble recombinant proteins, we found that the mutant protein had reduced binding to susceptible cells compared to wild type and diminished ability to inhibit BVDV infection, suggesting a lower affinity for BVDV receptors. We then analyzed the effect of β-hairpin mutations in the context of recombinant viral particles. Mutant viruses recovered from cell culture supernatant after transfection of recombinant RNA had almost completely inhibited ability to re-infect susceptible cells, indicating an impact of mutations on BVDV infectivity. Finally, sequential passaging of the mutant virus resulted in the selection of a viral population in which β-hairpin mutations reverted to the wild type sequence to restore infectivity. Taken together, our results show that this conserved region of the E2 protein is critical for the interaction with host cell receptors.
Main Authors: | , , , , , |
---|---|
Format: | info:ar-repo/semantics/artículo biblioteca |
Language: | eng |
Published: |
MDPI
2021-06
|
Subjects: | Enfermedades de los Animales, Pestivirus de la Diarrea Bovina, Proteínas Recombinantes, Animal Diseases, Bovine Diarrhoea Pestivirus, Pestivirus, Recombinant Proteins, Proteína E2, Protein E2, BVDV, |
Online Access: | http://hdl.handle.net/20.500.12123/10881 https://www.mdpi.com/1999-4915/13/6/1157 https://doi.org/10.3390/v13061157 |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
id |
oai:localhost:20.500.12123-10881 |
---|---|
record_format |
koha |
spelling |
oai:localhost:20.500.12123-108812021-12-10T13:34:34Z A β-Hairpin Motif in the Envelope Protein E2 Mediates Receptor Binding of Bovine Viral Diarrhea Virus Merwaiss, Fernando Pascual, María José Pomilio, María Trinidad Lopez, Maria Gabriela Taboga, Oscar Alberto Alvarez, Diego Ezequiel Enfermedades de los Animales Pestivirus de la Diarrea Bovina Proteínas Recombinantes Animal Diseases Bovine Diarrhoea Pestivirus Pestivirus Recombinant Proteins Proteína E2 Protein E2 BVDV Pestivirus envelope protein E2 is crucial to virus infection and accomplishes virus-receptor interaction during entry. However, mapping of E2 residues mediating these interactions has remained unexplored. In this study, to investigate the structure-function relationship for a β-hairpin motif exposed to the solvent in the crystal structure of bovine viral diarrhea virus (BVDV) E2, we designed two amino acidic substitutions that result in a change of electrostatic potential. First, using wild type and mutant E2 expressed as soluble recombinant proteins, we found that the mutant protein had reduced binding to susceptible cells compared to wild type and diminished ability to inhibit BVDV infection, suggesting a lower affinity for BVDV receptors. We then analyzed the effect of β-hairpin mutations in the context of recombinant viral particles. Mutant viruses recovered from cell culture supernatant after transfection of recombinant RNA had almost completely inhibited ability to re-infect susceptible cells, indicating an impact of mutations on BVDV infectivity. Finally, sequential passaging of the mutant virus resulted in the selection of a viral population in which β-hairpin mutations reverted to the wild type sequence to restore infectivity. Taken together, our results show that this conserved region of the E2 protein is critical for the interaction with host cell receptors. Instituto de Biotecnología Fil: Merwaiss, Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Merwaiss, Fernando. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Pascual, María José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Pascual, María José. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Pomilio, María Trinidad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Pomilio, María Trinidad. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Lopez, Maria Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Lopez, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina Fil: Taboga, Oscar Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Alvarez, Diego Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Alvarez, Diego Ezequiel. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina 2021-12-10T13:22:22Z 2021-12-10T13:22:22Z 2021-06 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/10881 https://www.mdpi.com/1999-4915/13/6/1157 1999-4915 https://doi.org/10.3390/v13061157 eng info:eu-repo/semantics/openAccess application/pdf MDPI Viruses 13 (6) : 1157 (2021) |
institution |
INTA AR |
collection |
DSpace |
country |
Argentina |
countrycode |
AR |
component |
Bibliográfico |
access |
En linea |
databasecode |
dig-inta-ar |
tag |
biblioteca |
region |
America del Sur |
libraryname |
Biblioteca Central del INTA Argentina |
language |
eng |
topic |
Enfermedades de los Animales Pestivirus de la Diarrea Bovina Proteínas Recombinantes Animal Diseases Bovine Diarrhoea Pestivirus Pestivirus Recombinant Proteins Proteína E2 Protein E2 BVDV Enfermedades de los Animales Pestivirus de la Diarrea Bovina Proteínas Recombinantes Animal Diseases Bovine Diarrhoea Pestivirus Pestivirus Recombinant Proteins Proteína E2 Protein E2 BVDV |
spellingShingle |
Enfermedades de los Animales Pestivirus de la Diarrea Bovina Proteínas Recombinantes Animal Diseases Bovine Diarrhoea Pestivirus Pestivirus Recombinant Proteins Proteína E2 Protein E2 BVDV Enfermedades de los Animales Pestivirus de la Diarrea Bovina Proteínas Recombinantes Animal Diseases Bovine Diarrhoea Pestivirus Pestivirus Recombinant Proteins Proteína E2 Protein E2 BVDV Merwaiss, Fernando Pascual, María José Pomilio, María Trinidad Lopez, Maria Gabriela Taboga, Oscar Alberto Alvarez, Diego Ezequiel A β-Hairpin Motif in the Envelope Protein E2 Mediates Receptor Binding of Bovine Viral Diarrhea Virus |
description |
Pestivirus envelope protein E2 is crucial to virus infection and accomplishes virus-receptor interaction during entry. However, mapping of E2 residues mediating these interactions has remained unexplored. In this study, to investigate the structure-function relationship for a β-hairpin motif exposed to the solvent in the crystal structure of bovine viral diarrhea virus (BVDV) E2, we designed two amino acidic substitutions that result in a change of electrostatic potential. First, using wild type and mutant E2 expressed as soluble recombinant proteins, we found that the mutant protein had reduced binding to susceptible cells compared to wild type and diminished ability to inhibit BVDV infection, suggesting a lower affinity for BVDV receptors. We then analyzed the effect of β-hairpin mutations in the context of recombinant viral particles. Mutant viruses recovered from cell culture supernatant after transfection of recombinant RNA had almost completely inhibited ability to re-infect susceptible cells, indicating an impact of mutations on BVDV infectivity. Finally, sequential passaging of the mutant virus resulted in the selection of a viral population in which β-hairpin mutations reverted to the wild type sequence to restore infectivity. Taken together, our results show that this conserved region of the E2 protein is critical for the interaction with host cell receptors. |
format |
info:ar-repo/semantics/artículo |
topic_facet |
Enfermedades de los Animales Pestivirus de la Diarrea Bovina Proteínas Recombinantes Animal Diseases Bovine Diarrhoea Pestivirus Pestivirus Recombinant Proteins Proteína E2 Protein E2 BVDV |
author |
Merwaiss, Fernando Pascual, María José Pomilio, María Trinidad Lopez, Maria Gabriela Taboga, Oscar Alberto Alvarez, Diego Ezequiel |
author_facet |
Merwaiss, Fernando Pascual, María José Pomilio, María Trinidad Lopez, Maria Gabriela Taboga, Oscar Alberto Alvarez, Diego Ezequiel |
author_sort |
Merwaiss, Fernando |
title |
A β-Hairpin Motif in the Envelope Protein E2 Mediates Receptor Binding of Bovine Viral Diarrhea Virus |
title_short |
A β-Hairpin Motif in the Envelope Protein E2 Mediates Receptor Binding of Bovine Viral Diarrhea Virus |
title_full |
A β-Hairpin Motif in the Envelope Protein E2 Mediates Receptor Binding of Bovine Viral Diarrhea Virus |
title_fullStr |
A β-Hairpin Motif in the Envelope Protein E2 Mediates Receptor Binding of Bovine Viral Diarrhea Virus |
title_full_unstemmed |
A β-Hairpin Motif in the Envelope Protein E2 Mediates Receptor Binding of Bovine Viral Diarrhea Virus |
title_sort |
β-hairpin motif in the envelope protein e2 mediates receptor binding of bovine viral diarrhea virus |
publisher |
MDPI |
publishDate |
2021-06 |
url |
http://hdl.handle.net/20.500.12123/10881 https://www.mdpi.com/1999-4915/13/6/1157 https://doi.org/10.3390/v13061157 |
work_keys_str_mv |
AT merwaissfernando abhairpinmotifintheenvelopeproteine2mediatesreceptorbindingofbovineviraldiarrheavirus AT pascualmariajose abhairpinmotifintheenvelopeproteine2mediatesreceptorbindingofbovineviraldiarrheavirus AT pomiliomariatrinidad abhairpinmotifintheenvelopeproteine2mediatesreceptorbindingofbovineviraldiarrheavirus AT lopezmariagabriela abhairpinmotifintheenvelopeproteine2mediatesreceptorbindingofbovineviraldiarrheavirus AT tabogaoscaralberto abhairpinmotifintheenvelopeproteine2mediatesreceptorbindingofbovineviraldiarrheavirus AT alvarezdiegoezequiel abhairpinmotifintheenvelopeproteine2mediatesreceptorbindingofbovineviraldiarrheavirus AT merwaissfernando bhairpinmotifintheenvelopeproteine2mediatesreceptorbindingofbovineviraldiarrheavirus AT pascualmariajose bhairpinmotifintheenvelopeproteine2mediatesreceptorbindingofbovineviraldiarrheavirus AT pomiliomariatrinidad bhairpinmotifintheenvelopeproteine2mediatesreceptorbindingofbovineviraldiarrheavirus AT lopezmariagabriela bhairpinmotifintheenvelopeproteine2mediatesreceptorbindingofbovineviraldiarrheavirus AT tabogaoscaralberto bhairpinmotifintheenvelopeproteine2mediatesreceptorbindingofbovineviraldiarrheavirus AT alvarezdiegoezequiel bhairpinmotifintheenvelopeproteine2mediatesreceptorbindingofbovineviraldiarrheavirus |
_version_ |
1756008233480224768 |