Ratification vote on taxonomic proposals to the International Committee on Taxonomy of Viruses (2012)

Adams, Michael J.; Carstens, Eric B.

doi:10.1007/s00705-012-1299-6

Cited by 289 publications

(186 citation statements)

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“…Additionally, herpesviral DNA is consistently detected in tumor tissues, but less commonly in nontumor tissues, from turtles from Hawaii (19), Florida (20), and Brazil (21). In 2012, the virus associated with FP, chelonid herpesvirus 5 (ChHV5), was designated the type for the newly erected genus Scutavirus of the subfamily Alphaherpesvirinae, family Herpesviridae (22).…”

mentioning

confidence: 99%

In Vitro Replication of Chelonid Herpesvirus 5 in Organotypic Skin Cultures from Hawaiian Green Turtles (Chelonia mydas)

Work

Dagenais

Weatherby

et al. 2017

J Virol

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Fibropapillomatosis (FP) is a tumor disease of marine turtles associated with chelonid herpesvirus 5 (ChHV5), which has historically been refractory to growth in tissue culture. Here we show, for the first time, de novo formation of ChHV5-positive intranuclear inclusions in cultured green turtle cells, which is indicative of active lytic replication of the virus. The minimal requirements to achieve lytic replication in cultured cells included (i) either in vitro cultures of ChHV5-positive tumor biopsy specimens (plugs) or organotypic cultures (rafts) consisting of ChHV5-positive turtle fibroblasts in collagen rafts seeded with turtle keratinocytes and (ii) keratinocyte maturation induced by raising raft or biopsy cultures to the air-liquid interface. Virus growth was confirmed by detailed electron microscopic studies that revealed intranuclear sun-shaped capsid factories, tubules, various stages of capsid formation, nuclear export by budding into the perinuclear space, tegument formation, and envelopment to complete de novo virus production. Membrane synthesis was also observed as a sign of active viral replication. Interestingly, cytoplasmic particles became associated with keratin filaments, a feature not seen in conventional monolayer cell cultures, in which most studies of herpesvirus replication have been performed. Our findings draw a rich and realistic picture of ChHV5 replication in cells derived from its natural host and may be crucial not only to better understand ChHV5 circulation but also to eventually complete Koch's postulates for FP. Moreover, the principles described here may serve as a model for culture of other viruses that are resistant to replication in conventional cell culture.IMPORTANCE A major challenge in virology is the study of viruses that cannot be grown in the laboratory. One example is chelonid herpesvirus 5 (ChHV5), which is associated with fibropapillomatosis, a globally distributed, debilitating, and fatal tumor disease of endangered marine turtles. Pathological examination shows that ChHV5 is shed in skin. Here we show that ChHV5 will grow in vitro if we replicate the complex three-dimensional structure of turtle skin. Moreover, lytic virus growth requires a close interplay between fibroblasts and keratinocytes. Finally, the morphogenesis of herpesviral growth in three-dimensional cultures reveals a far richer, and likely more realistic, array of capsid morphologies than that encountered in traditional monolayer cell cultures. Our findings have applications to other viruses, including those of humans.KEYWORDS herpesvirus, electron microscopy, immunohistochemistry, morphogenesis, pathobiology

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confidence: 99%

In Vitro Replication of Chelonid Herpesvirus 5 in Organotypic Skin Cultures from Hawaiian Green Turtles (Chelonia mydas)

Work

Dagenais

Weatherby

et al. 2017

J Virol

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“…Members of the CoVs are further classified into four genera: Alphacoronavirus, Betacoronavirus, Gammacoronavirus, and Deltacoronavirus (2). Human coronavirus OC43 (HCoV-OC43) was isolated from a patient with upper respiratory tract disease in the 1960s and classified into the Betacoronavirus genus (3).…”

mentioning

confidence: 99%

The ns12.9 Accessory Protein of Human Coronavirus OC43 Is a Viroporin Involved in Virion Morphogenesis and Pathogenesis

Wang

Ping

et al. 2015

J Virol

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An accessory gene between the S and E gene loci is contained in all coronaviruses (CoVs), and its function has been studied in some coronaviruses. This gene locus in human coronavirus OC43 (HCoV-OC43) encodes the ns12.9 accessory protein; however, its function during viral infection remains unknown. Here, we engineered a recombinant mutant virus lacking the ns12.9 protein (HCoV-OC43-⌬ns12.9) to characterize the contributions of ns12.9 in HCoV-OC43 replication. The ns12.9 accessory protein is a transmembrane protein and forms ion channels in both Xenopus oocytes and yeast through homo-oligomerization, suggesting that ns12.9 is a newly recognized viroporin. HCoV-OC43-⌬ns12.9 presented at least 10-fold reduction of viral titer in vitro and in vivo. Intriguingly, exogenous ns12.9 and heterologous viroporins with ion channel activity could compensate for the production of HCoV-OC43-⌬ns12.9, indicating that the ion channel activity of ns12.9 plays a significant role in the production of infectious virions. Systematic dissection of single-cycle replication revealed that ns12.9 protein had no measurable effect on virus entry, subgenomic mRNA (sgmRNA) synthesis, and protein expression. Further characterization revealed that HCoV-OC43-⌬ns12.9 was less efficient in virion morphogenesis than recombinant wild-type virus (HCoV-OC43-WT). Moreover, reduced viral replication, inflammatory response, and virulence in HCoV-OC43-⌬ns12.9-infected mice were observed compared to the levels for HCoV-OC43-WT-infected mice. Taken together, our results demonstrated that the ns12.9 accessory protein functions as a viroporin and is involved in virion morphogenesis and the pathogenesis of HCoV-OC43 infection. IMPORTANCEHCoV-OC43 was isolated in the 1960s and is a major agent of the common cold. The functions of HCoV-OC43 structural proteins have been well studied, but few studies have focused on its accessory proteins. In the present study, we demonstrated that the ns12.9 protein is a newly recognized viroporin, and the ns12.9 gene knockout virus (HCoV-OC43-⌬ns12.9) presents a growth defect in vitro and in vivo. We identified the important functions of the ns12.9 viroporin in virion morphogenesis during HCoV-OC43 infection. Furthermore, mice infected with HCoV-OC43-⌬ns12.9 exhibited reduced inflammation and virulence accompanied by a lower titer in the brain than that of wild-type-infected mice, suggesting the ns12.9 viroporin influences virus pathogenesis. Therefore, our findings revealed that the ns12.9 viroporin facilitates virion morphogenesis to enhance viral production, and these results provided a deeper understanding of HCoV-OC43 pathogenesis.T he coronaviruses (CoVs) belong to the Coronaviridae family of the order Nidovirales and are distributed widely among animals, birds, and humans (1). Members of the CoVs are further classified into four genera: Alphacoronavirus, Betacoronavirus, Gammacoronavirus, and Deltacoronavirus (2). Human coronavirus OC43 (HCoV-OC43) was isolated from a patient with upper respiratory tract disease ...

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“…However, with the expansion of intensive culture, various diseases have occurred frequently and severely affected production. Among the causes of these diseases, mud crab dicistrovirus-1 (MCDV-1) possesses Q3 a linear positive-sense ssRNA genome with a length of 10,415 nucleotides excluding the 3 poly(A) tail; MCDV-1 has been officially named mud crab virus (MCV) in 2012 by the International Committee on Taxonomy of Viruses (Adams and Carstens (2012)). This virus usually accompanies the mud crab reovirus (MCRV) in mud crabs, and can cause 100% mortality alone under artificial infection.…”

Section: Introductionmentioning

confidence: 99%

Rapid detection of mud crab dicistrovirus-1 using loop-mediated isothermal amplification

Guo¹,

Zhang²,

Ma³

et al. 2014

Journal of Virological Methods

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Ratification vote on taxonomic proposals to the International Committee on Taxonomy of Viruses (2012)

Cited by 289 publications

References 0 publications

In Vitro Replication of Chelonid Herpesvirus 5 in Organotypic Skin Cultures from Hawaiian Green Turtles (Chelonia mydas)

In Vitro Replication of Chelonid Herpesvirus 5 in Organotypic Skin Cultures from Hawaiian Green Turtles (Chelonia mydas)

The ns12.9 Accessory Protein of Human Coronavirus OC43 Is a Viroporin Involved in Virion Morphogenesis and Pathogenesis

Rapid detection of mud crab dicistrovirus-1 using loop-mediated isothermal amplification

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