Identification and characterization of the structural and nonstructural proteins of African horsesickness virus and determination of the genome coding assignments.
Abstract: Proteins present in purified African horsesickness virus (AHSV) and in infected cells were analyzed by SDS-polyacrylamide gel electrophoresis. Twelve viral proteins were identified, one minor and four major structural proteins, three major and two minor nonstructural proteins, as well as variable amounts of two additional structural proteins. Cell-free translation of total AHS virion RNA in a rabbit reticulocyte system resulted in the synthesis of proteins which were qualitatively and quantitatively similar to those found in infected cells. The in vivo and in vitro synthesized proteins were viral specific as demonstrated by immunoprecipitation. The coding assignments of all the purified genome segments were determined by in vitro translation and confirmed by immunoprecipitation.
Publication Date: 1992-02-01 PubMed ID: 1531096DOI: 10.1016/0042-6822(92)90009-eGoogle Scholar: Lookup
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Summary
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The study investigates the varying proteins present in the African horsesickness virus (AHSV) and in cells infected by this virus. A total of twelve viral proteins were identified and the coding assignments of all purified genome segments were determined through in vitro translation.
Types of Proteins in AHSV and Infected Cells
In this study, the researchers analyzed both the African horsesickness virus and cells that were infected by it. Samples were studied using SDS-polyacrylamide gel electrophoresis, a method for separating proteins by their molecular weight. Through this analysis:
- Twelve viral proteins were identified. These include one minor and four major structural proteins, three major and two minor nonstructural proteins, and a varying amount of two additional structural proteins.
Cell-free Translation of AHSV RNA
The researchers used a rabbit reticulocyte system for cell-free translation of the whole AHSV virion RNA. This process produces proteins that are qualitatively and quantitatively similar to those observed in the infected cells. The results indicate:
- That the synthesized proteins are specific to the virus, as confirmed by a technique known as immunoprecipitation. Immunoprecipitation is a method used to isolate a particular protein out of a solution.
Determination of Coding Assignments
Finally, the report explains how the coding assignments of the purified genome segments were determined. The study achieved this:
- By using in vitro translation, a method that allows the synthesis of large amounts of protein in a controlled environment outside of a cell.
- The assignments obtained through in vitro translation were then confirmed using immunoprecipitation.
This process allowed the researchers to assign specific functions to segments of the genetic material (RNA) of the AHSV, enhancing our understanding of how this virus operates at a molecular level.
Cite This Article
APA
Grubman MJ, Lewis SA.
(1992).
Identification and characterization of the structural and nonstructural proteins of African horsesickness virus and determination of the genome coding assignments.
Virology, 186(2), 444-451.
https://doi.org/10.1016/0042-6822(92)90009-e Publication
Researcher Affiliations
- USDA, ARS, NAA, Plum Island Animal Disease Center, Greenport, New York 11944.
MeSH Terms
- African Horse Sickness Virus / chemistry
- African Horse Sickness Virus / genetics
- Animals
- Capsid / analysis
- Capsid / genetics
- Cell Line
- Cricetinae
- Electrophoresis, Polyacrylamide Gel
- Genome, Viral
- Horses
- Vero Cells
- Viral Core Proteins / analysis
- Viral Core Proteins / genetics
- Viral Nonstructural Proteins
- Viral Proteins / analysis
- Viral Proteins / genetics
- Viral Structural Proteins / analysis
- Viral Structural Proteins / genetics
Citations
This article has been cited 13 times.- Dennis SJ, Meyers AE, Hitzeroth II, Rybicki EP. African Horse Sickness: A Review of Current Understanding and Vaccine Development. Viruses 2019 Sep 11;11(9).
- Ngoveni HG, van Schalkwyk A, Koekemoer JJO. Evidence of Intragenic Recombination in African Horse Sickness Virus. Viruses 2019 Jul 18;11(7).
- Schade-Weskott ML, van Schalkwyk A, Koekemoer JJO. A correlation between capsid protein VP2 and the plaque morphology of African horse sickness virus in cell culture. Virus Genes 2018 Aug;54(4):527-535.
- de la Poza F, Calvo-Pinilla E, López-Gil E, Marín-López A, Mateos F, Castillo-Olivares J, Lorenzo G, Ortego J. Ns1 is a key protein in the vaccine composition to protect Ifnar(-/-) mice against infection with multiple serotypes of African horse sickness virus. PLoS One 2013;8(7):e70197.
- Manole V, Laurinmäki P, Van Wyngaardt W, Potgieter CA, Wright IM, Venter GJ, van Dijk AA, Sewell BT, Butcher SJ. Structural insight into African horsesickness virus infection. J Virol 2012 Aug;86(15):7858-66.
- Thompson GM, Jess S, Murchie AK. A review of African horse sickness and its implications for Ireland. Ir Vet J 2012 Jul 5;65(1):9.
- Wilson A, Mellor PS, Szmaragd C, Mertens PP. Adaptive strategies of African horse sickness virus to facilitate vector transmission. Vet Res 2009 Mar-Apr;40(2):16.
- Firth AE. Bioinformatic analysis suggests that the Orbivirus VP6 cistron encodes an overlapping gene. Virol J 2008 Apr 14;5:48.
- Lord CC, Barnard B, Day K, Hargrove JW, McNamara JJ, Paul RE, Trenholme K, Woolhouse ME. Aggregation and distribution of strains in microparasites. Philos Trans R Soc Lond B Biol Sci 1999 Apr 29;354(1384):799-807.
- Stäuber N, Martinez-Costas J, Sutton G, Monastyrskaya K, Roy P. Bluetongue virus VP6 protein binds ATP and exhibits an RNA-dependent ATPase function and a helicase activity that catalyze the unwinding of double-stranded RNA substrates. J Virol 1997 Oct;71(10):7220-6.
- Stone-Marschat M, Carville A, Skowronek A, Laegreid WW. Detection of African horse sickness virus by reverse transcription-PCR. J Clin Microbiol 1994 Mar;32(3):697-700.
- Sakamoto K, Punyahotra R, Mizukoshi N, Ueda S, Imagawa H, Sugiura T, Kamada M, Fukusho A. Rapid detection of African horsesickness virus by the reverse transcriptase polymerase chain reaction (RT-PCR) using the amplimer for segment 3 (VP3 gene). Arch Virol 1994;136(1-2):87-97.
- van Staden V, Stoltz MA, Huismans H. Expression of nonstructural protein NS3 of African horsesickness virus (AHSV): evidence for a cytotoxic effect of NS3 in insect cells, and characterization of the gene products in AHSV infected Vero cells. Arch Virol 1995;140(2):289-306.
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