Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1997 Jul;71(7):5673–5677. doi: 10.1128/jvi.71.7.5673-5677.1997

Fish nodavirus lytic cycle and semipermissive expression in mammalian and fish cell cultures.

C Delsert 1, N Morin 1, M Comps 1
PMCID: PMC191815  PMID: 9188647

Abstract

In this study, Dicentrarchus labrax encephalitis virus (DlEV), which causes sea bass encephalitis, was propagated in cell culture, thus allowing study of its lytic cycle. DlEV infection of mammalian and fish cells induced different patterns of expression of capsid proteins, which were assembled as virus-like particles, accumulating in the cytoplasm either as diffuse masses or in vesicles, as shown by electron microscopy. These particles correspond to virions, as shown by their ability to induce secondary infection. Fish cells proved to be more permissive for DlEV than mammalian cells, although virus yield remained low. RNA analysis of infected sea bass cells revealed DlEV RNA3, in addition to genomic RNA1 and RNA2, and the presence of the RNA2 minus strand, thus demonstrating the replication of the DlEV genome. In addition, DlEV RNA-dependent RNA polymerase was associated with mature virions even after purification by a CsCl gradient, but it was dissociated when capsids were destabilized. In addition to providing more information about the relatedness of DlEV to the members of the family Nodaviridae, this study shows that fish nodaviruses may not be able to infect as wide a variety of cells as insect nodaviruses can.

Full Text

The Full Text of this article is available as a PDF (747.1 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  2. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  3. Delsert C. D., Rosenfeld M. G. A tissue-specific small nuclear ribonucleoprotein and the regulated splicing of the calcitonin/calcitonin gene-related protein transcript. J Biol Chem. 1992 Jul 25;267(21):14573–14579. [PubMed] [Google Scholar]
  4. Delsert C., D'Halluin J. C. Genetic expression of human adenoviruses in simian cells. Evidence for interserotypic inhibition of viral DNA synthesis. Virus Res. 1984;1(5):365–380. doi: 10.1016/0168-1702(84)90024-8. [DOI] [PubMed] [Google Scholar]
  5. Gallagher T. M., Friesen P. D., Rueckert R. R. Autonomous replication and expression of RNA 1 from black beetle virus. J Virol. 1983 May;46(2):481–489. doi: 10.1128/jvi.46.2.481-489.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gallagher T. M., Rueckert R. R. Assembly-dependent maturation cleavage in provirions of a small icosahedral insect ribovirus. J Virol. 1988 Sep;62(9):3399–3406. doi: 10.1128/jvi.62.9.3399-3406.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Longworth J. F., Carey G. P. A small RNA virus with a divided genome from Heteronychus arator (F.) [Coleoperai Scarabaeidae]. J Gen Virol. 1976 Oct;33(1):31–40. doi: 10.1099/0022-1317-33-1-31. [DOI] [PubMed] [Google Scholar]
  8. Mori K., Nakai T., Muroga K., Arimoto M., Mushiake K., Furusawa I. Properties of a new virus belonging to nodaviridae found in larval striped jack (Pseudocaranx dentex) with nervous necrosis. Virology. 1992 Mar;187(1):368–371. doi: 10.1016/0042-6822(92)90329-n. [DOI] [PubMed] [Google Scholar]
  9. Nishizawa T., Mori K., Furuhashi M., Nakai T., Furusawa I., Muroga K. Comparison of the coat protein genes of five fish nodaviruses, the causative agents of viral nervous necrosis in marine fish. J Gen Virol. 1995 Jul;76(Pt 7):1563–1569. doi: 10.1099/0022-1317-76-7-1563. [DOI] [PubMed] [Google Scholar]
  10. Selling B. H., Allison R. F., Kaesberg P. Genomic RNA of an insect virus directs synthesis of infectious virions in plants. Proc Natl Acad Sci U S A. 1990 Jan;87(1):434–438. doi: 10.1073/pnas.87.1.434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Selling B. H., Rueckert R. R. Plaque assay for black beetle virus. J Virol. 1984 Jul;51(1):251–253. doi: 10.1128/jvi.51.1.251-253.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES