scholarly journals Screening for the Proteins That Can Interact with Grouper Nervous Necrosis Virus Capsid Protein

Viruses ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 985
Author(s):  
Po-Yu Huang ◽  
Han-Chia Hsiao ◽  
Szu-Wen Wang ◽  
Shao-Fu Lo ◽  
Ming-Wei Lu ◽  
...  
Keyword(s):  
Optic Nerve ◽  
Capsid Protein ◽  
Protein Interactions ◽  
Ribosomal Proteins ◽  
Nervous Necrosis Virus ◽  
Structural Protein ◽  
Necrosis Virus ◽  
Ion Regulation ◽  
Protein Protein Interaction ◽  
Atp Generation

Nervous necrosis virus (NNV) can infect many species of fish and has an 80–100% mortality rate. NNV capsid protein (NNVCP) is the only structural protein of NNV, but there are few studies on the protein–protein interaction between NNVCP and the host cell. To investigate NNV morphogenesis, native NNV capsid protein (NNVCP) was used to screen for protein–protein interactions in this study. The results identified that 49 grouper optic nerve proteins can interact with NNVCP and may function as putative receptor or co-receptor, cytoskeleton, glucose metabolism and ATP generation, immunity, mitochondrial ion regulation, and ribosomal proteins. Creatine kinase B-type (CKB) is one of those 49 optic nerve proteins. CKB, a kind of enzyme of ATP generation, was confirmed to interact with NNVCP by far-Western blot and showed to colocalize with NNVCP in GF-1 cells. Compared to the control, the expression of CKB was significantly induced in the brain and eyes infected with NNV. Moreover, the amount of replication of NNV is relatively high in cells expressing CKB. In addition to providing the database of proteins that can interact with NNVCP for subsequent analysis, the results of this research also verified that CKB plays an important role in the morphogenesis of NNV.

In vitro neutralization of nervous necrosis virus by a nanobody binding to the protrusion domain of capsid protein

Aquaculture ◽  
2021 ◽  
pp. 737654
Author(s):  
Song Zhu ◽  
Bo Miao ◽  
Yu-Zhou Zhang ◽  
Wei-Wei Zeng ◽  
De-Shou Wang ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Nervous Necrosis Virus ◽  
Necrosis Virus

scholarly journals Proteomic Analysis of Antigen 60 Complex of M. bovis Bacillus Calmette-Guérin Reveals Presence of Extracellular Vesicle Proteins and Predicted Functional Interactions

Vaccines ◽  
2019 ◽  
Vol 7 (3) ◽  
pp. 80 ◽  
Author(s):  
Khayriyyah Mohd Hanafiah ◽  
Norsyahida Arifin ◽  
Paul R. Sanders ◽  
Nurulhasanah Othman ◽  
Mary L. Garcia ◽  
...  
Keyword(s):  
Western Blot ◽  
Protein Interactions ◽  
Ribosomal Proteins ◽  
Vaccine Development ◽  
Extracellular Vesicle ◽  
Biological Interactions ◽  
Bacillus Calmette Guerin ◽  
Protein Protein Interactions ◽  
Protein Protein Interaction ◽  
Antigen 60

Tuberculosis (TB) is ranked among the top 10 causes of death worldwide. New biomarker-based serodiagnostics and vaccines are unmet needs stalling disease control. Antigen 60 (A60) is a thermostable mycobacterial complex typically purified from Bacillus Calmette-Guérin (BCG) vaccine. A60 was historically evaluated for TB serodiagnostic and vaccine potential with variable findings. Despite containing immunogenic proteins, A60 has yet to be proteomically characterized. Here, commercial A60 was (1) trypsin-digested in-solution, analyzed by LC-MS/MS, searched against M. tuberculosis H37Rv and M. bovis BCG Uniprot databases; (2) analyzed using STRING to predict protein–protein interactions; and (3) probed with anti-TB monoclonal antibodies and patient immunoglobulin G (IgG) on Western blot to evaluate antigenicity. We detected 778 proteins in two A60 samples (440 proteins shared), including DnaK, LprG, LpqH, and GroEL1/2, reportedly present in mycobacterial extracellular vesicles (EV). Of these, 107 were also reported in EVs of M. tuberculosis, and 27 key proteins had significant protein–protein interaction, with clustering for chaperonins, ribosomal proteins, and proteins for ligand transport (LpqH and LprG). On Western blot, 7/8 TB and 1/8 non-TB sera samples had reactivity against 37–50 kDa proteins, while LpqH, GroEL2, and PstS1 were strongly detected. In conclusion, A60 comprises numerous proteins, including EV proteins, with predicted biological interactions, which may have implications on biomarker and vaccine development.

Assembly of the capsid protein of red-spotted grouper nervous necrosis virus during purification, and role of calcium ions in chromatography

2016 ◽  
Vol 21 (3) ◽  
pp. 373-380 ◽  
Author(s):  
Hyoung Jin Kim ◽  
Hye-Lim Kwag ◽  
Do Gyun Kim ◽  
Bo Kyu Kang ◽  
Sang Yoon Han ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Calcium Ions ◽  
Nervous Necrosis Virus ◽  
Necrosis Virus

NMR assignments of protrusion domain of capsid protein from dragon grouper nervous necrosis virus

2019 ◽  
Vol 14 (1) ◽  
pp. 63-66
Author(s):  
Petra Štěrbová ◽  
Danni Wu ◽  
Yuan-Chao Lou ◽  
Chun-Hsiung Wang ◽  
Wei-Hau Chang ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Nmr Assignments ◽  
Nervous Necrosis Virus ◽  
Necrosis Virus

scholarly journals Sequence of the non-structural protein gene encoded by RNA1 of striped jack nervous necrosis virus

1999 ◽  
Vol 80 (11) ◽  
pp. 3019-3022 ◽  
Author(s):  
Takahiro Nagai ◽  
Toyohiko Nishizawa
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequences ◽  
Nervous Necrosis Virus ◽  
Structural Protein ◽  
Necrosis Virus ◽  
Rna Molecules ◽  
Viral Nervous Necrosis ◽  
The Family ◽  
Amino Acid Levels ◽  
Structural Protein Gene

Striped jack nervous necrosis virus (SJNNV), the causative agent of viral nervous necrosis in marine fish, is a member of the family Nodaviridae whose genome consists of two positive-sense RNA molecules encapsidated in a single virion. In this study, the nucleotide sequence of SJNNV RNA1 was determined. The SJNNV RNA1 was 3081 bases long and contained a single ORF encoding 983 aa of approximately 110 kDa. The sequence identities between RNA1 of SJNNV and RNA1 of insect nodaviruses were 28% at the nucleotide and amino acid levels, although the conserved motifs for the RNA-dependent RNA polymerase were located at almost the same positions in the amino acid sequences. The present study, together with our previous work on SJNNV RNA2, suggests that a new genus, Piscinodavirus, should be created in the family Nodaviridae.

scholarly journals GHSC70 Is Involved in the Cellular Entry of Nervous Necrosis Virus

2014 ◽  
Vol 89 (1) ◽  
pp. 61-70 ◽  
Author(s):  
Jui-Shin Chang ◽  
Shau-Chi Chi
Keyword(s):  
Capsid Protein ◽  
Fish Species ◽  
Receptor Protein ◽  
Economic Losses ◽  
Nervous Necrosis Virus ◽  
Specific Receptor ◽  
Necrosis Virus ◽  
Content Type ◽  
Voltage Dependent ◽  
Selective Channel

ABSTRACTNervous necrosis virus (NNV) is a devastating pathogen of cultured marine fish and has affected more than 40 fish species. NNV belongs to the betanodaviruses ofNodaviridaeand is a nonenveloped icosahedral particle with 2 single-stranded positive-sense RNAs. To date, knowledge regarding NNV entry into the host cell remains limited, and no NNV-specific receptor protein has been published. Using grouper fin cell line GF-1 and purified NNV capsid protein in a virus overlay protein binding assay (VOPBA), grouper heat shock cognate protein 70 (GHSC70) and grouper voltage-dependent anion selective channel protein 2 (GVDAC2) were investigated as NNV receptor protein candidates. We cloned and sequenced the genes for GHSC70 and GVDAC2 and expressed them inEscherichia colifor antiserum preparation. Knockdown of the expression of GHSC70 and GVDAC2 genes with specific short interfering RNAs (siRNAs) significantly downregulated viral RNA expression in NNV-infected GF-1 cells. By performing an immunoprecipitation assay, we confirmed that GHSC70 interacted with NNV capsid protein, while VDAC2 did not. Immunofluorescence staining and flow cytometry analysis revealed the presence of the GHSC70 protein on the cell surface. After a blocking assay, we detected the NNV RNA2 levels after 1 h of adsorption to GF-1 cells; the level was significantly lower in the cells pretreated with the GHSC70 antiserum than in nontreated cells. Therefore, we suggest that GHSC70 participates in the NNV entry of GF-1 cells, likely functioning as an NNV receptor or coreceptor protein.IMPORTANCEFish nodavirus has caused mass mortality of more than 40 fish species worldwide and resulted in huge economic losses in the past 20 years. Among the four genotypes of fish nodaviruses, the red-spotted grouper nervous necrosis virus (RGNNV) genotype exhibits the widest host range. In our previous study, we developed monoclonal antibodies with high neutralizing efficiency against grouper NNV in GF-1 cells, indicating that NNV-specific receptor(s) may exist on the GF-1 cell membrane. However, no NNV receptor protein has been published. In this study, we found GHSC70 to be an NNV receptor (or coreceptor) candidate through VOBPA and provided several lines of evidence demonstrating that GHSC70 protein has a role in the NNV entry step of GF-1 cells. To the best of our knowledge, this is the first report identifying grouper HSC70 and its role in NNV entry into GF-1 cells.

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