Assembly of JC virus-like particles in COS7 cells

Shishido, Yukiko; Nukuzuma, Souichi; Mukaigawa, Jun; Morikawa, Shigeru; Yasui, Kohichiroh; Nagashima, Kazuo

doi:10.1002/(sici)1096-9071(199704)51:4<265::aid-jmv2>3.0.co;2-3

Cited by 18 publications

(12 citation statements)

References 28 publications

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“…Since establishment of a eukaryotic expression system to generate virus-like particles (VLPs) [119], we have been studying the molecular mechanisms underlying how JC virus develops intranuclear inclusions. The JC virus capsid would be composed of VP1, VP2, and VP3, likely in a specific ratio as in SV40 [80].…”

Section: The Jc Virus Biology and Molecular Pathogenesismentioning

confidence: 99%

Progressive multifocal leukoencephalopathy and promyelocytic leukemia nuclear bodies: a review of clinical, neuropathological, and virological aspects of JC virus-induced demyelinating disease

Shishido‐Hara

2010

Acta Neuropathol

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Progressive multifocal leukoencephalopathy is a fatal viral-induced demyelinating disease that was once rare but has become more prevalent today. Over the past decades, much has been learned about the disease from molecular study of the etiological agent of the disease, JC virus. Recently, promyelocytic leukemia nuclear bodies (PML-NBs), punctuate structures for important nuclear functions in eukaryotic cells, were identified as an intranuclear target of JC virus infection. Neuropathologically, JC virus-infected glial cells display diffuse amphophilic viral inclusions by hematoxylin–eosin staining (full inclusions), a diagnostic hallmark of this disease. Recent results using immunohistochemistry, however, revealed the presence of punctate viral inclusions preferentially located along the inner nuclear periphery (dot-shaped inclusions). Dot-shaped inclusions reflect the accumulation of viral progeny at PML-NBs, which may be disrupted after viral replication. Structural changes to PML-NBs have been reported for a variety of human diseases, including cancers and neurodegenerative disorders. Thus, PML-NBs may provide clues to the further pathogenesis of JC virus-induced demyelinating disease. Here, we review what we have learned since the disease entity establishment, including a look at recent progress in understanding the relationship between JC virus, etiology and PML-NBs.

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Section: The Jc Virus Biology and Molecular Pathogenesismentioning

confidence: 99%

Progressive multifocal leukoencephalopathy and promyelocytic leukemia nuclear bodies: a review of clinical, neuropathological, and virological aspects of JC virus-induced demyelinating disease

Shishido‐Hara

2010

Acta Neuropathol

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“…Three capsid proteins, the major capsid protein VP1 and minor capsid proteins VP2 and VP3, are encoded in an overlapping manner downstream of a small regulatory protein called agnoprotein. This viral genome was manipulated molecularly and inserted into a eukaryotic expression vector . The first system was established to produce JC virus‐like particles (VLPs) in cultured cells .…”

Section: Introductionmentioning

confidence: 99%

Progressive multifocal leukoencephalopathy: Dot‐shaped inclusions and virus‐host interactions

Shishido‐Hara

2015

Neuropathology

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Progressive multifocal leukoencephalopathy (PML) is a fatal demyelinating disease caused by reactivation of the asymptomatic persistent pathogen human polyomavirus JC (JC virus). The pathology of affected brain tissues demonstrates oligodendroglia-like cells with viral inclusions in their enlarged nuclei, a diagnostic hallmark of this disease. Today, the pathological features of this disease are expanding, partly due to an unsteady balance between viral virulence and host immunity. Intranuclear viral inclusions were initially thought to be amphophilic materials comprising the entire enlarged nucleus, based on HE staining (full inclusions). Howevewr, recent immunohistochemical analyses detected the presence of intranuclear viral inclusions in dots (dot-shaped inclusions). The dot-shaped inclusions reflect clustered progeny virions at punctuated subnuclear domains called promyelocytic leukemia nuclear bodies, and are indicative of early-stage viral infection or suppressed viral proliferation. Second, the JC virus is usually reactivated in patients with impaired immunity, and therefore the inflammatory reactions are poor. However, the causes of immunosuppression are divergent, as seen with the frequent use of immunosuppressive drugs, including natalizumab. Therefore, the degree of host immunity is variable; some patients show marked anti-viral inflammatory reactions and a good prognosis, indicating that a strong resistance against viral infection remains. Recovery of the immune system may also induce paradoxical clinical worsening, known as immune reconstitution inflammatory syndrome, the mechanism of which has not been clarified. The virus-host interactions have increased in complexity, and the pathology of PML is diverging. In this review, the pathology of PML will be described, with a focus on the intranuclear target of JC virus infection and host inflammatory reactions.

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“…In all cases, the presence of the VLPs has been demonstrated by using the hemagglutination assay or by transmission electron microscopy. 8,9,12,14 In this study, we wanted to determine the effect of the incubation buffer on the efficiency of VP1 VLPs assembly in cell extracts. For that reason, chicken RBCs were incubated with VP1 VLP containing samples produced in an E. coli wild type genetic background under different buffer conditions (Figure 1a).…”

Section: Resultsmentioning

confidence: 99%

“…[4][5][6][7] However, capsids formed exclusively of VP1 (hJCV VP1 virus-like particles-VLPs) can be obtained in heterologous expression systems including Escherichia coli, yeasts, mammalian cells, and insect cell-baculovirus expression systems. [8][9][10][11][12][13][14] An interesting feature of VLPs is their ability to self-assemble, which can be controlled experimentally allowing the internalization of dyes, nucleic acids, drugs, or proteins in vitro. 10,[15][16][17] In addition, VLPs can be functionalized with cell ligands allowing the specific delivery of the cargo to target cells.…”

Section: Introductionmentioning

confidence: 99%

Effect of the DnaK chaperone on the conformational quality of JCV VP1 virus‐like particles produced in Escherichia coli

Saccardo

Rodríguez‐Carmona

Villaverde

et al. 2014

Biotechnology Progress

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Protein nanoparticles such as virus-like particles (VLPs) can be obtained by recombinant protein production of viral capsid proteins and spontaneous self-assembling in cell factories. Contrarily to infective viral particles, VLPs lack infective viral genome while retaining important viral properties like cellular tropism and intracellular delivery of internalized molecules. These properties make VLPs promising and fully biocompatible nanovehicles for drug delivery. VLPs of human JC virus (hJCV) VP1 capsid protein produced in Escherichia coli elicit variable hemagglutination properties when incubated at different NaCl concentrations and pH conditions, being optimal at 200 mM NaCl and at pH range between 5.8 and 7.5. In addition, the presence or absence of chaperone DnaK in E. coli cells influence the solubility of recombinant VP1 and the conformational quality of this protein in the VLPs. The hemagglutination ability of hJCV VP1 VLPs contained in E. coli cell extracts can be modulated by buffer composition in the hemagglutination assay. It has been also determined that the production of recombinant hJCV VP1 in E. coli is favored by the absence of chaperone DnaK as observed by Western Blot analysis in different E. coli genetic backgrounds, indicating a proteolysis targeting role for DnaK. However, solubility is highly compromised in a DnaK(-) E. coli strain suggesting an important role of this chaperone in reduction of protein aggregates. Finally, hemagglutination efficiency of recombinant VP1 is directly related to the presence of DnaK in the producing cells.

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Assembly of JC virus-like particles in COS7 cells

Cited by 18 publications

References 28 publications

Progressive multifocal leukoencephalopathy and promyelocytic leukemia nuclear bodies: a review of clinical, neuropathological, and virological aspects of JC virus-induced demyelinating disease

Progressive multifocal leukoencephalopathy and promyelocytic leukemia nuclear bodies: a review of clinical, neuropathological, and virological aspects of JC virus-induced demyelinating disease

Progressive multifocal leukoencephalopathy: Dot‐shaped inclusions and virus‐host interactions

Effect of the DnaK chaperone on the conformational quality of JCV VP1 virus‐like particles produced in Escherichia coli

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