Purification and properties of virus particles, infectious subviral particles, cores and VP7 crystals of African horsesickness virus serotype 9

Burroughs, J. N.; Ohara, Reiko; Smale, C. J.; Hamblin, C.; Walton, Adam Z.; Armstrong, R.M.; Mertens, Peter P. C.

doi:10.1099/0022-1317-75-8-1849

Cited by 61 publications

(50 citation statements)

References 21 publications

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“…1C). Autonomous crys-tal formation in vivo is a feature unique to AHSV VP7 amongst the orbiviruses (Burroughs et al, 1994;Maree and Paweska, 2005). No similar structures to those described above were observed in wild-type baculovirus-infected cells (Fig.…”

Section: Synthesis and Characterization Of Ahsv Clps Composed Of Vp3 mentioning

confidence: 75%

“…1E), which is typical for authentic AHSV core particles (Fig. 1F), as described by Oellermann et al (1970) and Burroughs et al (1994). The icosahedral particles also displayed knob-like protrusions that radiated outwards, which is characteristic of the orbivirus ring-like arrange-ment of VP7 capsomers (Fig.…”

Section: Synthesis and Characterization Of Ahsv Clps Composed Of Vp3 mentioning

confidence: 83%

“…3A). A marked structural resemblance exists in the overall architecture of the major core proteins and core particles of these two related orbiviruses (Burroughs et al, 1994;Basak et al, 1996Basak et al, , 1997Manole et al, 2012), as well as acceptable phylogenetic relationships between their VP3 or VP7 proteins. The amino acid sequence identity shared between VP3 of AHSV-9 and BTV-1 was 57.5%, while their VP7 proteins shared 42.8% identity, thus exceeding the 30% cut-off value for homology modeling (Xiang, 2006).…”

Section: Engineering and Assembly Of Mutant Ahsv Clpsmentioning

confidence: 99%

“…The AHSV genome is composed of 10 segments of linear double-stranded (ds) RNA, which encode four nonstructural proteins (NS1 to NS4) and seven structural proteins (Bremer, 1976;Grubman and Lewis, 1992;Zwart et al, 2015). The virion is a nonenveloped icosahedral particle (Oellermann et al, 1970;Burroughs et al, 1994) composed of three concentric protein layers (Manole et al, 2012). The core particle comprises of a VP7 surface layer and an underlying VP3 layer, which encloses the genomic dsRNA segments and three enzymatic minor proteins (VP1, VP3 and VP6).…”

Section: Introductionmentioning

confidence: 99%

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Synthesis of empty african horse sickness virus particles

Maree

Putterill

et al. 2016

Virus Research

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As a means to develop African horse sickness (AHS) vaccines that are safe and DIVA compliant, we investigated the synthesis of empty African horse sickness virus (AHSV) particles. The emphasis of this study was on the assembly of the major viral core (VP3 and VP7) and outer capsid proteins (VP2 and VP5) into architecturally complex, heteromultimeric nanosized particles. The production of fully assembled corelike particles (CLPs) was accomplished in vivo by baculovirus-mediated co-synthesis of VP3 and VP7. The two different outer capsid proteins were capable of associating independently of each other with preformed cores to yield partial virus-like particles (VLPs). Complete VLPs were synthesized, albeit with a low yield. Crystalline formation of AHSV VP7 trimers is thought to impede high-level CLP production. Consequently, we engineered and co-synthesized VP3 with a more hydrophilic mutant VP7, resulting in an increase in the turnover of CLPs.

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Section: Synthesis and Characterization Of Ahsv Clps Composed Of Vp3 mentioning

confidence: 75%

Section: Synthesis and Characterization Of Ahsv Clps Composed Of Vp3 mentioning

confidence: 83%

Section: Engineering and Assembly Of Mutant Ahsv Clpsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Synthesis of empty african horse sickness virus particles

Maree

Putterill

et al. 2016

Virus Research

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“…When AHSV VP7 is expressed in mammalian or insect cells, the protein trimers localise to one or two sites in the cell and assemble into large flat hexagonal crystalline particles (Burroughs et al 1994;Venter et al 2014) which have recently been shown to impede the formation of core-like particles (Maree et al 2016). This characteristic is unique to AHSV, and presents a distinct difference between major capsid protein VP7 of AHSV and BTV (Monastyrskaya et al 1997).…”

Section: Discussionmentioning

confidence: 99%

A Dual Laser Scanning Confocal and Transmission Electron Microscopy Analysis of the Intracellular Localization, Aggregation and Particle Formation of African Horse Sickness Virus Major Core Protein VP7

et al. 2017

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The bulk of African horse sickness virus (AHSV) major core protein VP7 selfassembles into flat, hexagonal crystalline particles in a process appearing unrelated to viral replication. Why this unique characteristic of AHSV VP7 is genetically conserved, and whether VP7 aggregation and particle formation have an effect on cellular biology or the viral life cycle, is unknown. Here we investigated how different small peptide and enhanced green fluorescent protein (eGFP) insertions into the VP7 top domain affected VP7 localisation, aggregation and particle formation. This was done using a dual laser scanning confocal and transmission electron microscopy approach in conjunction with analyses of the solubility, aggregation and fluorescence profiles of the proteins. VP7 top domain modifications did not prevent trimerisation, or intracellular trafficking to one or two discrete sites in the cell. However, modifications that resulted in a misfolded and insoluble VP7-eGFP component blocked trafficking, and precluded protein accumulation at a single cellular site, perhaps by interfering with normal trimer-trimer interactions. Furthermore, the modifications disrupted the stable layering of the trimers into characteristic AHSV VP7 crystalline particles. It was concluded that VP7 trafficking is driven by a balance between VP7 solubility, trimer forming ability and trimer-trimer interactions.

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Utilization of cellulose microcapillary tubes as a model system for culturing and viral infection of mammalian cells

Venter

Merwe

Staden

2012

Microscopy Res & Technique

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Cryofixation by high-pressure freezing (HPF) and freeze substitution (FS) gives excellent preservation of intracellular membranous structures, ideal for ultrastructural investigations of virus infected cells. Conventional sample preparation methods of tissue cultured cells can however disrupt the association between neighbouring cells or of viruses with the plasma membrane, which impacts upon the effectiveness whereby virus release from cells can be studied. We established a system for virus infection and transmission electron microscopy preparation of mammalian cells that allowed optimal visualisation of membrane release events. African horse sickness virus (AHSV) is a non-enveloped virus that employs two different release mechanisms from mammalian 2 cells, i.e. lytic release through a disrupted plasma membrane and a non-lytic buddingtype release. Cellulose microcapillary tubes were used as support layer for culturing Vero cells. The cells grew to a confluent monolayer along the inside of the tubes and could readily be infected with AHSV. Sections of the microcapillary tubes proved easy to manipulate during the HPF procedure, showed no distortion or compression, and yielded well preserved cells in their native state. There was ample cell surface area available for visualisation, which allowed detection of both types of virus release at the plasma membrane at a significantly higher frequency than when utilising other methods.The consecutive culturing, virus infection and processing of cells within microcapillary tubes therefore represent a novel model system for monitoring intracellular virus life cycle and membrane release events, specifically suited to viruses that do not grow to high titres in tissue culture.

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Purification and properties of virus particles, infectious subviral particles, cores and VP7 crystals of African horsesickness virus serotype 9

Cited by 61 publications

References 21 publications

Synthesis of empty african horse sickness virus particles

Synthesis of empty african horse sickness virus particles

A Dual Laser Scanning Confocal and Transmission Electron Microscopy Analysis of the Intracellular Localization, Aggregation and Particle Formation of African Horse Sickness Virus Major Core Protein VP7

Utilization of cellulose microcapillary tubes as a model system for culturing and viral infection of mammalian cells

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