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Rescue of disabled infectious single-cycle (DISC) equine arteritis virus by using complementing cell lines that express minor structural glycoproteins.
Abstract: Equine arteritis virus (EAV) contains seven structural proteins that are all required to produce infectious progeny. Alphavirus-based expression vectors have been generated for each of these proteins to explore the possibilities for their constitutive expression in cell lines. This approach was successful for minor glycoproteins GP(2b), GP(3) and GP(4) and for the E protein. Subsequently, it was demonstrated that cell lines expressing these proteins could rescue EAV mutants that were disabled in the expression of the corresponding gene, resulting in the production of virus particles carrying the mutant genome. This system was particularly efficient for GP(2b)- and GP(4)-knockout mutants. Upon infection of non-complementing cells with these mutants, a self-limiting single cycle of replication was initiated, resulting in the expression of all but one of the viral proteins. These disabled infectious single-cycle (DISC) arteriviruses can also be used to express foreign sequences and are potentially useful in both fundamental research and vaccine development.
Publication Date: 2004-11-24 PubMed ID: 15557244DOI: 10.1099/vir.0.80443-0Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This research explores the use of cell lines expressing certain viral proteins to produce virus particles of the equine arteritis virus (EAV), specifically those that have random gene mutations. The successful implementation of the experiment could provide a basis for further research and potentially the development of vaccines.
Exploring Structural Proteins
- Equine arteritis virus (EAV) contains seven structural proteins – all of which are essential in producing infectious offspring.
- To further understand these proteins, the researchers used alphavirus-based expression vectors – these are a genetic tool used to make cells express a specific gene.
- By applying this to all seven proteins, it was found that minor glycoproteins GP(2b), GP(3) and GP(4) and the E protein were able to be expressed in cell lines.
Complementing Cell Lines
- The study established that cell lines which constitutively expressed these proteins were able to rescue EAV mutants that couldn’t express the corresponding gene.
- These mutants eventually produced virus particles that contained the mutant genome – a particularly successful result was found with the GP(2b) and GP(4) knockout mutants.
Disabled Infectious Single-Cycle (DISC) Arteriviruses
- When these mutated viruses were factored into non-complementing cells, it triggered a limited single cycle of replication, leading to the expression of all other viral proteins, except for one.
- The researchers identified these versions of the virus as disabled infectious single-cycle (DISC) arteriviruses.
- Due to their unique properties, DISC arteriviruses can serve as a good platform for expressing foreign sequences, which is beneficial for fundamental research.
- Besides, DISC arteriviruses contain the potential to be used in vaccine development due to their ability to produce offsprings with a partial viral genome.
Cite This Article
APA
Zevenhoven-Dobbe JC, Greve S, van Tol H, Spaan WJM, Snijder EJ.
(2004).
Rescue of disabled infectious single-cycle (DISC) equine arteritis virus by using complementing cell lines that express minor structural glycoproteins.
J Gen Virol, 85(Pt 12), 3709-3714.
https://doi.org/10.1099/vir.0.80443-0 Publication
Researcher Affiliations
- Molecular Virology Laboratory, Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, LUMC E4-P, Room P4-26, PO Box 9600, 2300 RC Leiden, The Netherlands.
- Molecular Virology Laboratory, Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, LUMC E4-P, Room P4-26, PO Box 9600, 2300 RC Leiden, The Netherlands.
- Molecular Virology Laboratory, Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, LUMC E4-P, Room P4-26, PO Box 9600, 2300 RC Leiden, The Netherlands.
- Molecular Virology Laboratory, Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, LUMC E4-P, Room P4-26, PO Box 9600, 2300 RC Leiden, The Netherlands.
- Molecular Virology Laboratory, Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, LUMC E4-P, Room P4-26, PO Box 9600, 2300 RC Leiden, The Netherlands.
MeSH Terms
- Animals
- Cell Line
- Cricetinae
- Defective Viruses / physiology
- Equartevirus / physiology
- Viral Structural Proteins / physiology
Citations
This article has been cited 10 times.- Vatter HA, Di H, Donaldson EF, Baric RS, Brinton MA. Each of the eight simian hemorrhagic fever virus minor structural proteins is functionally important. Virology 2014 Aug;462-463:351-62.
- Balasuriya UB, Zhang J, Go YY, MacLachlan NJ. Experiences with infectious cDNA clones of equine arteritis virus: lessons learned and insights gained. Virology 2014 Aug;462-463:388-403.
- Yun SI, Lee YM. Overview: Replication of porcine reproductive and respiratory syndrome virus. J Microbiol 2013 Dec;51(6):711-23.
- Balasuriya UB, Go YY, MacLachlan NJ. Equine arteritis virus. Vet Microbiol 2013 Nov 29;167(1-2):93-122.
- Tian D, Wei Z, Zevenhoven-Dobbe JC, Liu R, Tong G, Snijder EJ, Yuan S. Arterivirus minor envelope proteins are a major determinant of viral tropism in cell culture. J Virol 2012 Apr;86(7):3701-12.
- Firth AE, Zevenhoven-Dobbe JC, Wills NM, Go YY, Balasuriya UBR, Atkins JF, Snijder EJ, Posthuma CC. Discovery of a small arterivirus gene that overlaps the GP5 coding sequence and is important for virus production. J Gen Virol 2011 May;92(Pt 5):1097-1106.
- Tijms MA, Nedialkova DD, Zevenhoven-Dobbe JC, Gorbalenya AE, Snijder EJ. Arterivirus subgenomic mRNA synthesis and virion biogenesis depend on the multifunctional nsp1 autoprotease. J Virol 2007 Oct;81(19):10496-505.
- Posthuma CC, Nedialkova DD, Zevenhoven-Dobbe JC, Blokhuis JH, Gorbalenya AE, Snijder EJ. Site-directed mutagenesis of the Nidovirus replicative endoribonuclease NendoU exerts pleiotropic effects on the arterivirus life cycle. J Virol 2006 Feb;80(4):1653-61.
- Choi YJ, Yun SI, Kang SY, Lee YM. Identification of 5' and 3' cis-acting elements of the porcine reproductive and respiratory syndrome virus: acquisition of novel 5' AU-rich sequences restored replication of a 5'-proximal 7-nucleotide deletion mutant. J Virol 2006 Jan;80(2):723-36.
- Wissink EH, Kroese MV, van Wijk HA, Rijsewijk FA, Meulenberg JJ, Rottier PJ. Envelope protein requirements for the assembly of infectious virions of porcine reproductive and respiratory syndrome virus. J Virol 2005 Oct;79(19):12495-506.
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