In vitro assembly of polymorphic virus-like particles from the capsid protein of a nodavirus

Bajaj, Saumya; Banerjee, Manidipa

doi:10.1016/j.virol.2016.05.025

Cited by 20 publications

(26 citation statements)

References 40 publications

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“…Proof-ofprinciple measurements demonstrated the utility of the HIV-1 capsid biosensor for testing binding modes of host proteins involved in promoting HIV-1 infection, suggesting a role of oligomerization for CPSF6 binding. The underlying design can conceptually be extended to other viruses [45][46][47][48] for discovery and characterization of proteins, small molecules and drugs that interact with viral capsids to facilitate or inhibit gene delivery and viral replication.…”

Section: Discussionmentioning

confidence: 99%

Fluorescence biosensor for real-time interaction dynamics of host proteins with HIV-1 capsid tubes

Lau

Walsh

Wang

et al. 2019

Preprint

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The human immunodeficiency virus 1 (HIV-1) capsid serves as a binding platform for proteins and small molecules from the host cell that regulate various steps in the virus life cycle. However, there are currently no quantitative methods that use assembled capsid lattices for measuring host-pathogen interaction dynamics. Here we developed a single molecule fluorescence biosensor using self-assembled capsid tubes as biorecognition elements and imaged capsid binders using total internal reflection fluorescence microscopy in a microfluidic setup. The method is highly sensitive in its ability to observe and quantify binding, obtain dissociation constants, extract kinetics with an extended application of using more complex analytes that can accelerate characterisation of novel capsid binders.

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Section: Discussionmentioning

confidence: 99%

Fluorescence biosensor for real-time interaction dynamics of host proteins with HIV-1 capsid tubes

Lau

Walsh

Wang

et al. 2019

Preprint

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“…Chromy et al showed that chaperone proteins were needed for the in vitro assembly of polyomavirus VP1 into particles of uniform size in an energy‐dependent manner. A more recent study by Bajaj and Banerjee on the in vitro assembly of Flock House virus (FHV) VLPs revealed that the coat protein assembled into VLPs of various sizes in the absence of the chaperone and scaffolding proteins. This indicates that different hosts could have different systems for particle assembly, resulting in VLPs of different sizes.…”

Section: Discussionmentioning

confidence: 99%

Virus‐like particle of Macrobrachium rosenbergii nodavirus produced in Spodoptera frugiperda (Sf9) cells is distinctive from that produced in Escherichia coli

Kueh

Yong

Dezfooli

et al. 2016

Biotechnology Progress

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Macrobrachium rosenbergii nodavirus (MrNV) is a virus native to giant freshwater prawn. Recombinant MrNV capsid protein has been produced in Escherichia coli, which self-assembled into virus-like particles (VLPs). However, this recombinant protein is unstable, degrading and forming heterogenous VLPs. In this study, MrNV capsid protein was produced in insect Spodoptera frugiperda (Sf9) cells through a baculovirus system. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) revealed that the recombinant protein produced by the insect cells self-assembled into highly stable, homogenous VLPs each of approximately 40 nm in diameter. Enzyme-linked immunosorbent assay (ELISA) showed that the VLPs produced in Sf9 cells were highly antigenic and comparable to those produced in E. coli. In addition, the Sf9 produced VLPs were highly stable across a wide pH range (2-12). Interestingly, the Sf9 produced VLPs contained DNA of approximately 48 kilo base pairs and RNA molecules. This study is the first report on the production and characterization of MrNV VLPs produced in a eukaryotic system. The MrNV VLPs produced in Sf9 cells were about 10 nm bigger and had a uniform morphology compared with the VLPs produced in E. coli. The insect cell production system provides a good source of MrNV VLPs for structural and immunological studies as well as for host-pathogen interaction studies. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:549-557, 2017.

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“…DGNNV VLPs produced in E. coli were crystallized and studied with x-ray diffraction, revealing a T = 3 icosahedral structure approximately 38 nm in diameter, closely resembling the native virion ( Luo et al, 2014 ). Recombinant FHV capsid protein produced in E. coli was also used in an in vitro assembly study ( Bajaj & Banerjee, 2016 ). The capsid protein possesses additional N-terminal tag which hinders the assembly of the capsid protein into VLPs.…”

Section: Survey Methodologymentioning

confidence: 99%

Advances in the study of nodavirus

Yong

Yeap

Omar

et al. 2017

PeerJ

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Nodaviruses are small bipartite RNA viruses which belong to the family of Nodaviridae. They are categorized into alpha-nodavirus, which infects insects, and beta-nodavirus, which infects fishes. Another distinct group of nodavirus infects shrimps and prawns, which has been proposed to be categorized as gamma-nodavirus. Our current review focuses mainly on recent studies performed on nodaviruses. Nodavirus can be transmitted vertically and horizontally. Recent outbreaks have been reported in China, Indonesia, Singapore and India, affecting the aquaculture industry. It also decreased mullet stock in the Caspian Sea. Histopathology and transmission electron microscopy (TEM) are used to examine the presence of nodaviruses in infected fishes and prawns. For classification, virus isolation followed by nucleotide sequencing are required. In contrast to partial sequence identification, profiling the whole transcriptome using next generation sequencing (NGS) offers a more comprehensive comparison and characterization of the virus. For rapid diagnosis of nodavirus, assays targeting the viral RNA based on reverse-transcription PCR (RT-PCR) such as microfluidic chips, reverse-transcription loop-mediated isothermal amplification (RT-LAMP) and RT-LAMP coupled with lateral flow dipstick (RT-LAMP-LFD) have been developed. Besides viral RNA detections, diagnosis based on immunological assays such as enzyme-linked immunosorbent assay (ELISA), immunodot and Western blotting have also been reported. In addition, immune responses of fish and prawn are also discussed. Overall, in fish, innate immunity, cellular type I interferon immunity and humoral immunity cooperatively prevent nodavirus infections, whereas prawns and shrimps adopt different immune mechanisms against nodavirus infections, through upregulation of superoxide anion, prophenoloxidase, superoxide dismutase (SOD), crustin, peroxinectin, anti-lipopolysaccharides and heat shock proteins (HSP). Potential vaccines for fishes and prawns based on inactivated viruses, recombinant proteins or DNA, either delivered through injection, oral feeding or immersion, are also discussed in detail. Lastly, a comprehensive review on nodavirus virus-like particles (VLPs) is presented. In recent years, studies on prawn nodavirus are mainly focused on Macrobrachium rosenbergii nodavirus (MrNV). Recombinant MrNV VLPs have been produced in prokaryotic and eukaryotic expression systems. Their roles as a nucleic acid delivery vehicle, a platform for vaccine development, a molecular tool for mechanism study and in solving the structures of MrNV are intensively discussed.

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In vitro assembly of polymorphic virus-like particles from the capsid protein of a nodavirus

Cited by 20 publications

References 40 publications

Fluorescence biosensor for real-time interaction dynamics of host proteins with HIV-1 capsid tubes

Fluorescence biosensor for real-time interaction dynamics of host proteins with HIV-1 capsid tubes

Virus‐like particle of Macrobrachium rosenbergii nodavirus produced in Spodoptera frugiperda (Sf9) cells is distinctive from that produced in Escherichia coli

Advances in the study of nodavirus

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