Generation of Tioman virus nucleocapsid-like particles in yeast Saccharomyces cerevisiae

Petraityte, Rasa; Tamošiūnas, Paulius Lukas; Juozapaitis, Mindaugas; Žvirblienė, Aurelija; Sasnauskas, Kęstutis; Shiell, Brian J.; Russell, Gail; Bingham, John; Michalski, Wojtek P.

doi:10.1016/j.virusres.2009.06.013

Cited by 14 publications

(8 citation statements)

References 31 publications

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“…Proteins were passively eluted twice with 0.1 % SDS in PBS at room temperature overnight and concentrated in an Amicon Ultra-0.5 ml centrifugal filter with a 3 kDa molecular mass cut-off (Merck Millipore). Protein quantification was then performed using a NanoDrop spectrophotometer (Thermo Fisher Scientific) and the identity of extracted proteins was confirmed by mass spectrometric analysis using the method described previously (Petraityte et al, 2009). The mass spectra were matched against a custom database of NBV protein sequences.…”

Section: Methodsmentioning

confidence: 99%

Mouse fibroblast L929 cells are less permissive to infection by Nelson Bay orthoreovirus compared to other mammalian cell lines

Mok

Wynne

Grimley

et al. 2015

Journal of General Virology

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In recent years, bats have been identified as a natural reservoir for a diverse range of viruses. Nelson Bay orthoreovirus (NBV) was first isolated from the heart blood of a fruit bat (Pteropus poliocephalus) in 1968. While the pathogenesis of NBV remains unknown, other related members of this group have caused acute respiratory disease in humans. Thus the potential for NBV to impact human health appears plausible. Here, to increase our knowledge of NBV, we examined the replication and infectivity of NBV using different mammalian cell lines derived from bat, human, mouse and monkey. All cell lines supported the replication of NBV; however, L929 cells showed a greater than 2 log reduction in virus titre compared with the other cell lines. Furthermore, NBV did not induce major cytopathic effects in the L929 cells, as was observed in other cell lines. Interestingly, the related Pteropine orthoreoviruses, Pulau virus (PulV) and Melaka virus (MelV) were able to replicate to high titres in L929 cells but infection resulted in reduced cytopathic effect. Our study demonstrates a unique virus-host interaction between NBV and L929 cells, where cells effectively control viral infection/replication and limit the formation of syncytia. By elucidating the molecular mechanisms that control this unique relationship, important insights will be made into the biology of this fusogenic virus.

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Section: Methodsmentioning

confidence: 99%

Mouse fibroblast L929 cells are less permissive to infection by Nelson Bay orthoreovirus compared to other mammalian cell lines

Mok

Wynne

Grimley

et al. 2015

Journal of General Virology

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“…Antibodies against Hendra virus, Cedar virus, Tioman virus, and Australian bat lyssavirus antigens were detected at the Australian Animal Health Laboratory in Geelong, Victoria using multiplex microsphere assays (Luminex, Austin, USA) as described previously [29]. The conformational status of the viruses used were the following; soluble native-like oligomeric G envelope glycoproteins of HeV and CedV (sG tet ) were produced from stable expressing FreeStyle TM 293F cell lines [45,46], Tioman virus was a nucleocapsid protein expressed in the yeast Saccharomyces cerevisiae [47], and Australian bat lyssavirus was a nucleocapsid protein prepared in E.coli [48]. Briefly, prior to analysis, serum samples were first heat treated at 56˚C for 30 minutes to inactivate complement then the assay proteins were coupled to individual microsphere bead sets, of predetermined numbers of magnetic beads, MagPlex 1 (Luminex, Northbrook, USA).…”

Section: Serology For Hendra Virus Cedar Virus Tioman Virus and Rabmentioning

confidence: 99%

Seroprevalence of three paramyxoviruses; Hendra virus, Tioman virus, Cedar virus and a rhabdovirus, Australian bat lyssavirus, in a range expanding fruit bat, the Grey-headed flying fox (Pteropus poliocephalus)

et al. 2020

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Habitat-mediated global change is driving shifts in species' distributions which can alter the spatial risks associated with emerging zoonotic pathogens. Many emerging infectious pathogens are transmitted by highly mobile species, including bats, which can act as spill-over hosts for pathogenic viruses. Over three years, we investigated the seroepidemiology of paramyxoviruses and Australian bat lyssavirus in a range-expanding fruit bat, the Greyheaded flying fox (Pteropus poliocephalus), in a new camp in Adelaide, South Australia. Over six, biannual, sampling sessions, we quantified median florescent intensity (MFI) antibody levels for four viruses for a total of 297 individual bats using a multiplex Luminex binding assay. Where appropriate, florescence thresholds were determined using finite mixture modelling to classify bats' serological status. Overall, apparent seroprevalence of antibodies directed at Hendra, Cedar and Tioman virus antigens was 43.2%, 26.6% and 95.7%, respectively. We used hurdle models to explore correlates of seropositivity and antibody levels when seropositive. Increased body condition was significantly associated with Hendra seropositivity (Odds ratio = 3.67; p = 0.002) and Hendra virus levels were significantly higher in pregnant females (p = 0.002). While most bats were seropositive for Tioman virus, antibody levels for this virus were significantly higher in adults (p < 0.001). Unexpectedly, all sera were negative for Australian bat lyssavirus. Temporal variation in antibody levels suggests that antibodies to Hendra virus and Tioman virus may wax and wane on a seasonal basis. These findings suggest a common exposure to Hendra virus and other paramyxoviruses in this flying fox camp in South Australia.

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“…Establishment, validation and comparison of a PUUVBawa N protein-based indirect IgG and capture ELISA Purified, yeast-expressed viral antigens have broadly been used for diagnostics of virus infections in humans and animals [76][77][78][79][80][81]. We have recently described indirect and mAb-capture IgG-ELISAs for the detection of human infections with SNV, ANDV, PUUV, DOBV and HTNV [32,[36][37][38].…”

Section: Reactivity Of Human Sera and Mabs With Different Puuv N Antimentioning

confidence: 99%

Phylogenetic analysis of Puumala virus subtype Bavaria, characterization and diagnostic use of its recombinant nucleocapsid protein

et al. 2011

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Puumala virus (PUUV) is the predominant hantavirus species in Germany causing large numbers of mild to moderate cases of haemorrhagic fever with renal syndrome (HFRS). During an outbreak in South-East Germany in 2004 a novel PUUV subtype designated Bavaria was identified as the causative agent of HFRS in humans [1]. Here we present a molecular characterization of this PUUV strain by investigating novel partial and almost entire nucleocapsid (N) protein-encoding small (S-) segment sequences and partial medium (M-) segment sequences from bank voles (Myodes glareolus) trapped in Lower Bavaria during 2004 and 2005. Phylogenetic analyses confirmed their classification as subtype Bavaria, which is further subdivided into four geographical clusters. The entire N protein, harbouring an amino-terminal hexahistidine tag, of the Bavarian strain was produced in yeast Saccharomyces cerevisiae and showed a slightly different reactivity with N-specific monoclonal antibodies, compared to the yeast-expressed N protein of the PUUV strain Vranica/Hällnäs. Endpoint titration of human sera from different parts of Germany and from Finland revealed only very slight differences in the diagnostic value of the different recombinant proteins. Based on the novel N antigen indirect and monoclonal antibody capture IgG-ELISAs were established. By using serum panels from Germany and Finland their validation demonstrated a high sensitivity and specificity. In summary, our investigations demonstrated the Bavarian PUUV strain to be genetically divergent from other PUUV strains and the potential of its N protein for diagnostic applications.

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Generation of Tioman virus nucleocapsid-like particles in yeast Saccharomyces cerevisiae

Cited by 14 publications

References 31 publications

Mouse fibroblast L929 cells are less permissive to infection by Nelson Bay orthoreovirus compared to other mammalian cell lines

Mouse fibroblast L929 cells are less permissive to infection by Nelson Bay orthoreovirus compared to other mammalian cell lines

Seroprevalence of three paramyxoviruses; Hendra virus, Tioman virus, Cedar virus and a rhabdovirus, Australian bat lyssavirus, in a range expanding fruit bat, the Grey-headed flying fox (Pteropus poliocephalus)

Phylogenetic analysis of Puumala virus subtype Bavaria, characterization and diagnostic use of its recombinant nucleocapsid protein

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