scholarly journals Typing of viral hemorrhagic septicemia virus by monoclonal antibodies

2012 ◽  
Vol 93 (12) ◽  
pp. 2546-2557 ◽  
Author(s):  
Takafumi Ito ◽  
Jun Kurita ◽  
Motohiko Sano ◽  
Helle Frank Skall ◽  
Niels Lorenzen ◽  
...  
Keyword(s):  
Antigenic Determinants ◽  
Viral Hemorrhagic Septicemia Virus ◽  
N Protein ◽  
Viral Hemorrhagic Septicemia ◽  
Genotype Iii ◽  
P Protein ◽  
The Usa ◽  
Nucleoprotein N ◽  
Genotype Ivb ◽  
Reaction Patterns

Seven mAbs with specific reaction patterns against each of the four genotypes and eight subtypes of viral hemorrhagic septicemia virus (VHSV) were produced, aiming to establish an immunoassay for typing VHSV isolates according to their genotype. Among the mAbs, VHS-1.24 reacted with all genotypes except genotype Ie, whilst mAb VHS-9.23 reacted with all genotypes except genotype III. mAb VHS-3.80 reacted with genotypes Ib, Ic, Id and II. mAb VHS-7.57 reacted with genotypes II and IVa, and mAb VHS-5.18 with genotype Ib only. Interestingly, mAb VHS-3.75 reacted with all of the genotype III isolates except a rainbow trout-pathogenic isolate from the west coast of Norway, and reacted in addition with the IVb isolate, CA-NB00-01, from the east coast of the USA. Finally, mAb VHS-1.88 reacted with all genotype IVb isolates from the Great Lakes, but not with CA-NB00-01. In conclusion, we can distinguish between all four genotypes and between five of eight subtypes of VHSV by testing isolates in immunoassay using a panel of nine mAbs. By Western blotting and transfection of cell cultures, it was shown that mAb VHS-1.24 recognized an epitope on the viral phosphoprotein (P), whilst all others recognized antigenic determinants on the nucleoprotein (N). From amino acid alignments of the various genotypes and subtypes of VHSV isolates, it was possible to determine the epitope specificity of mAb VHS-1.24 to be aa 32–34 in the P-protein; the specificities of mAbs VHS-3.80, VHS-7.57 and VHS-3.75 were found to be aa 43 and 45–48, aa 117 and 121, and aa 103, 118 and 121 of the N-protein, respectively.

scholarly journals Fin and gill biopsies are effective nonlethal samples for detection of Viral hemorrhagic septicemia virus genotype IVb

2013 ◽  
Vol 25 (2) ◽  
pp. 203-209 ◽  
Author(s):  
Emily R. Cornwell ◽  
Chelsea A. Bellmund ◽  
Geoffrey H. Groocock ◽  
Po Ting Wong ◽  
Katherine L. Hambury ◽  
...  
Keyword(s):  
Viral Hemorrhagic Septicemia Virus ◽  
Virus Genotype ◽  
Viral Hemorrhagic Septicemia ◽  
Genotype Ivb

scholarly journals Role of Intermolecular Interactions of Vesicular Stomatitis Virus Nucleoprotein in RNA Encapsidation

2007 ◽  
Vol 82 (2) ◽  
pp. 674-682 ◽  
Author(s):  
Xin Zhang ◽  
Todd J. Green ◽  
Jun Tsao ◽  
Shihong Qiu ◽  
Ming Luo
Keyword(s):  
Crystal Structure ◽  
Intermolecular Interactions ◽  
Vesicular Stomatitis Virus ◽  
N Protein ◽  
P Protein ◽  
Intermolecular Contacts ◽  
Vesicular Stomatitis ◽  
Nucleoprotein N ◽  
Stable Ring

ABSTRACT The crystal structure of the vesicular stomatitis virus nucleoprotein (N) in complex with RNA reveals extensive and specific intermolecular interactions among the N molecules in the 10-member oligomer. What roles these interactions play in encapsidating RNA was studied by mutagenesis of the N protein. Three N mutants intended for disruption of the intermolecular interactions were designed and coexpressed with the phosphoprotein (P) in an Escherichia coli system previously described (T. J. Green et al., J. Virol. 74:9515-9524, 2000). Mutants N (Δ1-22), N (Δ347-352), and N (320-324, (Ala)5) lost RNA encapsidation and oligomerization but still bound with P. Another mutant, N (Ser290→Trp), was able to form a stable ring-like N oligomer and bind with the P protein but was no longer able to encapsidate RNA. The crystal structure of N (Ser290→Trp) at 2.8 Å resolution showed that this mutant can maintain all the same intermolecular interactions as the wild-type N except for a slight unwinding of the N-terminal lobe. These results suggest that the intermolecular contacts among the N molecules are required for encapsidation of the viral RNA.

scholarly journals Definition of the immune evasion-replication interface of rabies virus P protein

2021 ◽  
Vol 17 (7) ◽  
pp. e1009729
Author(s):  
Jingyu Zhan ◽  
Angela R. Harrison ◽  
Stephanie Portelli ◽  
Thanh Binh Nguyen ◽  
Isshu Kojima ◽  
...  
Keyword(s):  
Rabies Virus ◽  
Immune Evasion ◽  
Structural Data ◽  
Direct Analysis ◽  
Multifunctional Protein ◽  
N Protein ◽  
Binding Interface ◽  
P Protein ◽  
Definition Of ◽  
Nucleoprotein N

Rabies virus phosphoprotein (P protein) is a multifunctional protein that plays key roles in replication as the polymerase cofactor that binds to the complex of viral genomic RNA and the nucleoprotein (N protein), and in evading the innate immune response by binding to STAT transcription factors. These interactions are mediated by the C-terminal domain of P (PCTD). The colocation of these binding sites in the small globular PCTD raises the question of how these interactions underlying replication and immune evasion, central to viral infection, are coordinated and, potentially, coregulated. While direct data on the binding interface of the PCTD for STAT1 is available, the lack of direct structural data on the sites that bind N protein limits our understanding of this interaction hub. The PCTD was proposed to bind via two sites to a flexible loop of N protein (Npep) that is not visible in crystal structures, but no direct analysis of this interaction has been reported. Here we use Nuclear Magnetic Resonance, and molecular modelling to show N protein residues, Leu381, Asp383, Asp384 and phosphor-Ser389, are likely to bind to a ‘positive patch’ of the PCTD formed by Lys211, Lys214 and Arg260. Furthermore, in contrast to previous predictions we identify a single site of interaction on the PCTD by this Npep. Intriguingly, this site is proximal to the defined STAT1 binding site that includes Ile201 to Phe209. However, cell-based assays indicate that STAT1 and N protein do not compete for P protein. Thus, it appears that interactions critical to replication and immune evasion can occur simultaneously with the same molecules of P protein so that the binding of P protein to activated STAT1 can potentially occur without interrupting interactions involved in replication. These data suggest that replication complexes might be directly involved in STAT1 antagonism.

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