Three-dimensional reconstruction of baculovirus expressed bluetongue virus core-like particles by cryo-electron microscopy

Hewat, E.A.; Booth, Timothy F.; Loudon, Peter T.; Roy, Polly

doi:10.1016/0042-6822(92)90676-g

Cited by 95 publications

(68 citation statements)

References 22 publications

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“…This outer layer is probably composed of VP7 trimers as for BTV (Grimes et al, 1995). The CLP diameter of approximately 72 nm is in agreement with the diameter of BTV CLPs (Hewat et al, 1992) obtained from cryoelectron microscopic images, which is slightly greater (4 %) than the diameter of the authentic core (Burroughs et al, 1994). The identity of the CLPs was further verified by means of antibody decorating using VP7 monoclonal antibodies.…”

Section: Ddfsupporting

confidence: 70%

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Intracellular production of African horsesickness virus core-like particles by expression of the two major core proteins, VP3 and VP7, in insect cells.

Maree

Durbach

Huismans

1998

Journal of General Virology

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To gain more insight into the structure of the African horsesickness virus (AHSV) core particle, we have cloned, partially characterized and expressed the two major core proteins, VP3 and VP7, of AHSV-9. VP7 was found to be highly conserved amongst different serotypes. The VP3 and VP7 genes were subsequently expressed in insect cells by means of recombinant baculoviruses. VP7 was synthesized to very high levels and aggregated into distinctive crystals. Co-expression of VP3 and VP7 resulted in the intracellular formation of core-like particles which structurally resembled empty AHSV cores.African horsesickness is a highly infectious non-contagious disease of equines and in horses the mortality rate exceeds 90 %. The aetiological agent, African horsesickness virus (AHSV), belongs to the genus Orbivirus in the family Reoviridae. Like bluetongue virus (BTV), the prototype orbivirus, the AHSV virion is composed of seven structural proteins organized into a double-layered capsid containing the 10 double-stranded (ds) RNA segments of the viral genome (Bremer, 1976 ;Roy et al., 1994). The icosahedral core is composed of two major proteins, VP3 and VP7, and three minor structural proteins (VP1, 4 and 6). It has been shown that VP7 trimers constitute the surface of the BTV core : hence the prominent knob-like protrusions which overlay a scaffold of VP3 (Prasad et al., 1992 ;Grimes et al., 1995). The outer capsid layer is composed of the two major proteins, VP2 and VP5, of which VP2 is the serotype-specific antigen (Burrage et al

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Section: Ddfsupporting

confidence: 70%

“…2 b). Data obtained by Hewat et al (1992) suggest that BTV CLPs differ from the authentic cores by the absence of VP7 spikes around the fivefold axis, thereby reducing the knobby appearance. Liu et al (1992) reported that in vitro assembled CLPs displayed even less surface structure than BTV CLPs, perhaps due to incomplete VP7 incorporation.…”

Section: Ddfmentioning

confidence: 99%

Intracellular production of African horsesickness virus core-like particles by expression of the two major core proteins, VP3 and VP7, in insect cells.

Maree

Durbach

Huismans

1998

Journal of General Virology

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“…In this regard, it is worth noting that BTV VP7 trimers are soluble and do not aggregate into crystalline structures. High yields of BTV CLPs produced in the baculovirus expression system are typically reported (French and Roy, 1990;Hewat et al, 1992;Belyaev and Roy, 1993;Roy et al, 1997).…”

Section: Discussionmentioning

confidence: 99%

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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“…The genome is enclosed within the core particle, which comprises two major proteins, VP3 and 7 which are highly conserved among the nine AHSV serotypes, and three minor proteins, VP 1, 4 and 6 [20,100,108]. Together they make up the group specific epitopes [42,52,105]. The core particle is surrounded by the outer capsid composed of two proteins VP2 and 5.…”

Section: Aetiologymentioning

confidence: 99%

African horse sickness

2004

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-African horse sickness virus (AHSV) causes a non-contagious, infectious insect-borne disease of equids and is endemic in many areas of sub-Saharan Africa and possibly Yemen in the Arabian Peninsula. However, periodically the virus makes excursions beyond its endemic areas and has at times extended as far as India and Pakistan in the east and Spain and Portugal in the west. The vectors are certain species of Culicoides biting midge the most important of which is the AfroAsiatic species C. imicola. This paper describes the effects that AHSV has on its equid hosts, aspects of its epidemiology, and present and future prospects for control. The distribution of AHSV seems to be governed by a number of factors including the efficiency of control measures, the presence or absence of a long term vertebrate reservoir and, most importantly, the prevalence and seasonal incidence of the major vector which is controlled by climate. However, with the advent of climatechange the major vector, C. imicola, has now significantly extended its range northwards to include much of Portugal, Spain, Italy and Greece and has even been recorded from southern Switzerland. Furthermore, in many of these new locations the insect is present and active throughout the entire year. With the related bluetongue virus, which utilises the same vector species of Culicoides this has, since 1998, precipitated the worst outbreaks of bluetongue disease ever recorded with the virus extending further north in Europe than ever before and apparently becoming endemic in that continent. The prospects for similar changes in the epidemiology and distribution of AHSV are discussed.

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Three-dimensional reconstruction of baculovirus expressed bluetongue virus core-like particles by cryo-electron microscopy

Cited by 95 publications

References 22 publications

Intracellular production of African horsesickness virus core-like particles by expression of the two major core proteins, VP3 and VP7, in insect cells.

Intracellular production of African horsesickness virus core-like particles by expression of the two major core proteins, VP3 and VP7, in insect cells.

Synthesis of empty african horse sickness virus particles

African horse sickness

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