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Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation

Pancreas disease (PD) and sleeping disease (SD) are important viral scourges in aquaculture of Atlantic salmon and rainbow trout. The etiological agent of PD and SD is salmonid alphavirus (SAV), an unusual member of the Togaviridae (genus Alphavirus). SAV replicates at lower temperatures in fish. Ou...

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Autores principales: Metz, Stefan W., Feenstra, Femke, Villoing, Stephane, van Hulten, Marielle C., van Lent, Jan W., Koumans, Joseph, Vlak, Just M., Pijlman, Gorben P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3185042/
https://www.ncbi.nlm.nih.gov/pubmed/21991361
http://dx.doi.org/10.1371/journal.pone.0025816
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author Metz, Stefan W.
Feenstra, Femke
Villoing, Stephane
van Hulten, Marielle C.
van Lent, Jan W.
Koumans, Joseph
Vlak, Just M.
Pijlman, Gorben P.
author_facet Metz, Stefan W.
Feenstra, Femke
Villoing, Stephane
van Hulten, Marielle C.
van Lent, Jan W.
Koumans, Joseph
Vlak, Just M.
Pijlman, Gorben P.
author_sort Metz, Stefan W.
collection PubMed
description Pancreas disease (PD) and sleeping disease (SD) are important viral scourges in aquaculture of Atlantic salmon and rainbow trout. The etiological agent of PD and SD is salmonid alphavirus (SAV), an unusual member of the Togaviridae (genus Alphavirus). SAV replicates at lower temperatures in fish. Outbreaks of SAV are associated with large economic losses of ∼17 to 50 million $/year. Current control strategies rely on vaccination with inactivated virus formulations that are cumbersome to obtain and have intrinsic safety risks. In this research we were able to obtain non-infectious virus-like particles (VLPs) of SAV via expression of recombinant baculoviruses encoding SAV capsid protein and two major immunodominant viral glycoproteins, E1 and E2 in Spodoptera frugiperda Sf9 insect cells. However, this was only achieved when a temperature shift from 27°C to lower temperatures was applied. At 27°C, precursor E2 (PE2) was misfolded and not processed by host furin into mature E2. Hence, E2 was detected neither on the surface of infected cells nor as VLPs in the culture fluid. However, when temperatures during protein expression were lowered, PE2 was processed into mature E2 in a temperature-dependent manner and VLPs were abundantly produced. So, temperature shift-down during synthesis is a prerequisite for correct SAV glycoprotein processing and recombinant VLP production.
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spelling pubmed-31850422011-10-11 Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation Metz, Stefan W. Feenstra, Femke Villoing, Stephane van Hulten, Marielle C. van Lent, Jan W. Koumans, Joseph Vlak, Just M. Pijlman, Gorben P. PLoS One Research Article Pancreas disease (PD) and sleeping disease (SD) are important viral scourges in aquaculture of Atlantic salmon and rainbow trout. The etiological agent of PD and SD is salmonid alphavirus (SAV), an unusual member of the Togaviridae (genus Alphavirus). SAV replicates at lower temperatures in fish. Outbreaks of SAV are associated with large economic losses of ∼17 to 50 million $/year. Current control strategies rely on vaccination with inactivated virus formulations that are cumbersome to obtain and have intrinsic safety risks. In this research we were able to obtain non-infectious virus-like particles (VLPs) of SAV via expression of recombinant baculoviruses encoding SAV capsid protein and two major immunodominant viral glycoproteins, E1 and E2 in Spodoptera frugiperda Sf9 insect cells. However, this was only achieved when a temperature shift from 27°C to lower temperatures was applied. At 27°C, precursor E2 (PE2) was misfolded and not processed by host furin into mature E2. Hence, E2 was detected neither on the surface of infected cells nor as VLPs in the culture fluid. However, when temperatures during protein expression were lowered, PE2 was processed into mature E2 in a temperature-dependent manner and VLPs were abundantly produced. So, temperature shift-down during synthesis is a prerequisite for correct SAV glycoprotein processing and recombinant VLP production. Public Library of Science 2011-10-03 /pmc/articles/PMC3185042/ /pubmed/21991361 http://dx.doi.org/10.1371/journal.pone.0025816 Text en Metz 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, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Metz, Stefan W.
Feenstra, Femke
Villoing, Stephane
van Hulten, Marielle C.
van Lent, Jan W.
Koumans, Joseph
Vlak, Just M.
Pijlman, Gorben P.
Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation
title Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation
title_full Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation
title_fullStr Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation
title_full_unstemmed Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation
title_short Low Temperature-Dependent Salmonid Alphavirus Glycoprotein Processing and Recombinant Virus-Like Particle Formation
title_sort low temperature-dependent salmonid alphavirus glycoprotein processing and recombinant virus-like particle formation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3185042/
https://www.ncbi.nlm.nih.gov/pubmed/21991361
http://dx.doi.org/10.1371/journal.pone.0025816
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