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 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.

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.

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.

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