Immunofluorescent detection of double-stranded RNA in cells infected with reovirus, infectious pancreatic necrosis virus, and infectious bursal disease virus

1980 ◽  
Vol 26 (2) ◽  
pp. 256-261 ◽  
Author(s):  
Richard D. Macdonald
Keyword(s):  
Infectious Bursal Disease Virus ◽  
Pancreatic Necrosis ◽  
Infectious Pancreatic Necrosis Virus ◽  
Necrosis Virus ◽  
Urea Treatment ◽  
Double Stranded Rna ◽  
Bursal Disease ◽  
Infected Cells ◽  
Infectious Pancreatic Necrosis ◽  
Pancreatic Necrosis Virus

Urea treatment of ethanol-fixed virus-infected cells exposed nucleic acid antigens for immunofluorescence. Three double-stranded (ds) RNA-containing viruses showed bright fluorescence using antibodies against dsRNA. Three single-stranded RNA-containing viruses showed less intense fluorescence with anti-dsRNA. Four out of five cell lines persistently infected with various RNA-containing viruses showed no dsRNA detectable by immunofluorescence.

Intracellular replication of infectious pancreatic necrosis virus

1972 ◽  
Vol 18 (6) ◽  
pp. 865-867 ◽  
Author(s):  
Jeanne Argot ◽  
R. G. Malsberger
Keyword(s):  
Rna Synthesis ◽  
Deoxyribonucleic Acid ◽  
Pancreatic Necrosis ◽  
Infectious Pancreatic Necrosis Virus ◽  
Green Fluorescence ◽  
Necrosis Virus ◽  
Double Stranded Rna ◽  
Infectious Pancreatic Necrosis ◽  
Pancreatic Necrosis Virus ◽  
Viral Inclusions

Deoxyribonucleic acid (DNA) synthesis was repressed in cells 4 h after infection with infectious pancreatic necrosis (IPN) virus as determined by scintillation counting. Ribonucleic acid (RNA) synthesis was stimulated beginning 4 to 5 h after infection and reached a peak at 6 h. No Feulgen-positive viral inclusions were observed and the occasional inclusions seen exhibited green fluorescence when stained with acridine orange. These results suggest that the nucleic acid of IPN is double-stranded RNA.

scholarly journals VP3, a Structural Protein of Infectious Pancreatic Necrosis Virus, Interacts with RNA-Dependent RNA Polymerase VP1 and with Double-Stranded RNA

2007 ◽  
Vol 81 (12) ◽  
pp. 6652-6663 ◽  
Author(s):  
Torunn Pedersen ◽  
Astrid Skjesol ◽  
Jorunn B. Jørgensen
Keyword(s):  
Amino Acids ◽  
Pancreatic Necrosis ◽  
Infectious Pancreatic Necrosis Virus ◽  
Necrosis Virus ◽  
Interaction Domain ◽  
Particle Assembly ◽  
Double Stranded Rna ◽  
Terminal Amino ◽  
Infectious Pancreatic Necrosis ◽  
Pancreatic Necrosis Virus

ABSTRACT Infectious pancreatic necrosis virus (IPNV) is a bisegmented, double-stranded RNA (dsRNA) virus of the Birnaviridae family that causes widespread disease in salmonids. Its two genomic segments are encapsulated together with the viral RNA-dependent RNA polymerase, VP1, and the assumed internal protein, VP3, in a single-shell capsid composed of VP2. Major aspects of the molecular biology of IPNV, such as particle assembly and interference with host macromolecules, are as yet poorly understood. To understand the infection process, analysis of viral protein interactions is of crucial importance. In this study, we focus on the interaction properties of VP3, the suggested key organizer of particle assembly in birnaviruses. By applying the yeast two-hybrid system in combination with coimmunoprecipitation, VP3 was proven to bind to VP1 and to self-associate strongly. In addition, VP3 was shown to specifically bind to dsRNA in a sequence-independent manner by in vitro pull-down experiments. The binding between VP3 and VP1 was not dependent on the presence of dsRNA. Deletion analyses mapped the VP3 self-interaction domain within the 101 N-terminal amino acids and the VP1 interaction domain within the 62 C-terminal amino acids of VP3. The C-terminal end was also crucial but not sufficient for the dsRNA binding capacity of VP3. For VP1, the 90 C-terminal amino acids constituted the only dispensable part for maintaining VP3-binding ability. Kinetic analysis revealed the presence of VP1-VP3 complexes prior to the formation of mature virions in IPNV-infected CHSE-214 cells, which indicates a role in promoting the assembly process.

scholarly journals Genome Assembly and Particle Maturation of the Birnavirus Infectious Pancreatic Necrosis Virus

2004 ◽  
Vol 78 (24) ◽  
pp. 13829-13838 ◽  
Author(s):  
Rodrigo A. Villanueva ◽  
José L. Galaz ◽  
Juan A. Valdés ◽  
Matilde M. Jashés ◽  
Ana María Sandino
Keyword(s):  
Pancreatic Necrosis ◽  
Infectious Pancreatic Necrosis Virus ◽  
Necrosis Virus ◽  
Particle Assembly ◽  
Double Stranded Rna ◽  
Viral Particles ◽  
Agarose Electrophoresis ◽  
Electrophoresis System ◽  
Infectious Pancreatic Necrosis ◽  
Pancreatic Necrosis Virus

ABSTRACT In this study, we have analyzed the morphogenesis of the birnavirus infectious pancreatic necrosis virus throughout the infective cycle in CHSE-214 cells by using a native agarose electrophoresis system. Two types of viral particles (designated A and B) were identified, isolated, and characterized both molecularly and biologically. Together, our results are consistent with a model of morphogenesis in which the genomic double-stranded RNA is immediately assembled, after synthesis, into a large (66-nm diameter) and uninfectious particle A, where the capsid is composed of both mature and immature viral polypeptides. Upon maturation, particles A yield particles B through the proteolytic cleavage of most of the remaining viral precursors within the capsid, the compaction of the particle (60-nm diameter), and the acquisition of infectivity. These studies will provide the foundation for further analyses of birnavirus particle assembly and RNA replication.

scholarly journals Relationship between apoptosis and the BH2 domain sequence of the VP5 peptide of infectious pancreatic necrosis virus

2014 ◽  
pp. 3990-4002 ◽  
Author(s):  
Cesar Ortega S ◽  
Sylvia Rodríguez S ◽  
Juan Carlos Espinoza ◽  
Juan Kuznar ◽  
Alex Romero ◽  
...  
Keyword(s):  
Amino Acid ◽  
Pancreatic Necrosis ◽  
Virulent Strain ◽  
Critical Factor ◽  
Infectious Pancreatic Necrosis Virus ◽  
Viral Titer ◽  
Necrosis Virus ◽  
Infected Cells ◽  
Infectious Pancreatic Necrosis ◽  
Pancreatic Necrosis Virus

ABSTRACTObjective. To determine whether the level of apoptosis induced by infectious pancreatic necrosis virus (IPNV) is related to the amino acid sequence of the BH2 domain of the VP5 protein and the level of infectivity. Materials and methods. Three IPNV strains were used, the VP2 protein gene was amplified for genotyping and the VP5 sequence was also obtained. The infectivity of the strains was calculated using the viral titer obtained at 12, 24, 36 and 45 hpi in CHSE-214 cells. The percentage of apoptosis in infected cells was visualized by TUNEL assay and immunohistochemistry (caspase 3 detection). Results. The V70/06 and V33/98 strains corresponded to genotype Sp, while V112/06 to VR-299; the amino acid analysis of the V70/06 strain allows its classification as middle virulent strain and V33/98 and V112/06 strains as low virulent ones; infection with the V112/06 strain produced a lower viral titer (p<0.05). The VP5 gene of the 3 strains showed four homologous domains to Bcl-2, however, the BH2 domain was truncated in V70/06 and V33/98 (12 kDa), being complete (15kDa) in V112/06, which also showed the Trp155 residue, equivalent to Trp188 considered as a critical factor for the function of Bcl-2. The average apoptosis was below 12%, showing no differences between strains (p>0.05). Conclusions. The results showed that the differences in the BH2 sequence of the VP5 protein, infectivity and the VP2 sequence are not associated with the modulation of apoptosis.

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