Monoclonal Antibody 3F11 Against Structural Protein N in Equine Arteritis Virus

Hybridoma ◽  
2012 ◽  
Vol 31 (6) ◽  
pp. 479-479
Keyword(s):  
Monoclonal Antibody ◽  
Equine Arteritis Virus ◽  
Structural Protein

scholarly journals A NS1-binding monoclonal antibody interacts with two residues that are highly conserved in seasonal as well as newly emerged influenza A virus

2019 ◽  
Vol 77 (1) ◽  
Author(s):  
Su Hui Catherine Teo ◽  
Jian-Ping Wu ◽  
Chee-Keng Mok ◽  
Yee-Joo Tan
Keyword(s):  
Monoclonal Antibody ◽  
Influenza A Virus ◽  
Influenza A ◽  
Rna Binding ◽  
Intracellular Localization ◽  
Cross Reactivity ◽  
Structural Protein ◽  
Multifunctional Protein ◽  
Good Target ◽  
Conformational Plasticity

Abstract The non-structural protein 1 (NS1) of influenza A virus (IAV) is a multifunctional protein that antagonizes host antiviral responses, modulating virus pathogenesis. As such, it serves as a good target for research and diagnostic assay development. In this study, we have generated a novel monoclonal antibody (mAb) 19H9 and epitope mapping revealed that two residues, P85 and Y89, of NS1 are essential for interacting with this mAb. Furthermore, residues P85 and Y89 are found to be highly conserved across different IAV subtypes, namely seasonal H1N1 and H3N2, as well as the highly pathogenic H5N1 and H5N6 avian strains. Indeed, mAb 19H9 exhibits broad cross-reactivity with IAV strains of different subtypes. The binding of mAb 19H9 to residue Y89 was further confirmed by the abrogation of interaction between NS1 and p85β. Additionally, mAb 19H9 also detected NS1 proteins expressed in IAV-infected cells, showing NS1 intracellular localization in the cytoplasm and nucleolus. To our knowledge, mAb 19H9 is the first murine mAb to bind at the juxtaposition between the N-terminal RNA-binding domain and C-terminal effector domain of NS1. It could serve as a useful research tool for studying the conformational plasticity and dynamic changes in NS1.

scholarly journals Production of Recombinant EAV with Tagged Structural Protein Gp3 to Study Artervirus Minor Protein Localization in Infected Cells

Viruses ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 735
Author(s):  
Anna Karolina Matczuk ◽  
Grzegorz Chodaczek ◽  
Maciej Ugorski
Keyword(s):  
Protein Localization ◽  
Open Reading Frames ◽  
Equine Arteritis Virus ◽  
Structural Protein ◽  
N Protein ◽  
Virus Attachment ◽  
Recombinant Viruses ◽  
E Protein ◽  
Host Cell Factors ◽  
Minor Protein

Equine arteritis virus (EAV) is a prototype member of the Arterivirus family, comprising important pathogens of domestic animals. Minor glycoproteins of Arteriviruses are responsible for virus entry and cellular tropism. The experimental methods for studying minor Arterivirus proteins are limited because of the lack of antibodies and nested open reading frames (ORFs). In this study, we generated recombinant EAV with separated ORFs 3 and 4, and Gp3 carrying HA-tag (Gp3-HA). The recombinant viruses were stable on passaging and replicated in titers similar to the wild-type EAV. Gp3-HA was incorporated into the virion particles as monomers and as a Gp2/Gp3-HA/Gp4 trimer. Gp3-HA localized in ER and, to a lesser extent, in the Golgi, it also co-localized with the E protein but not with the N protein. The co-localization of Gp3-HA and the E protein with ERGIC was reduced. Moreover, EAV with Gp3-HA could become a valuable research tool for identifying host cell factors during infection and the role of Gp3 in virus attachment and entry.

scholarly journals Arterivirus Subgenomic mRNA Synthesis and Virion Biogenesis Depend on the Multifunctional nsp1 Autoprotease

2007 ◽  
Vol 81 (19) ◽  
pp. 10496-10505 ◽  
Author(s):  
Marieke A. Tijms ◽  
Danny D. Nedialkova ◽  
Jessika C. Zevenhoven-Dobbe ◽  
Alexander E. Gorbalenya ◽  
Eric J. Snijder
Keyword(s):  
Life Cycle ◽  
Rna Synthesis ◽  
Reverse Genetics ◽  
Rna Viruses ◽  
Regulatory Protein ◽  
Equine Arteritis Virus ◽  
Structural Protein ◽  
Mrna Synthesis ◽  
Nonstructural Proteins ◽  
Autocatalytic Processing

ABSTRACT Many groups of plus-stranded RNA viruses produce additional, subgenomic mRNAs to regulate the expression of part of their genome. Arteriviruses and coronaviruses (order Nidovirales) are unique among plus-stranded RNA viruses for using a mechanism of discontinuous RNA synthesis to produce a nested set of 5′- and 3′-coterminal subgenomic mRNAs, which serve to express the viral structural protein genes. The discontinuous step presumably occurs during minus-strand synthesis and joins noncontiguous sequences copied from the 3′- and 5′-proximal domains of the genomic template. Nidovirus genome amplification (“replication”) and subgenomic mRNA synthesis (“transcription”) are driven by 13 to 16 nonstructural proteins (nsp's), generated by autocatalytic processing of two large “replicase” polyproteins. Previously, using a replicon system, the N-terminal nsp1 replicase subunit of the arterivirus equine arteritis virus (EAV) was found to be dispensable for replication but crucial for transcription. Using reverse genetics, we have now addressed the role of nsp1 against the background of the complete EAV life cycle. Mutagenesis revealed that nsp1 is in fact a multifunctional regulatory protein. Its papain-like autoprotease domain releases nsp1 from the replicase polyproteins, a cleavage essential for viral RNA synthesis. Several mutations in the putative N-terminal zinc finger domain of nsp1 selectively abolished transcription, while replication was either not affected or even increased. Other nsp1 mutations did not significantly affect either replication or transcription but still dramatically reduced the production of infectious progeny. Thus, nsp1 is involved in at least three consecutive key processes in the EAV life cycle: replicase polyprotein processing, transcription, and virion biogenesis.

scholarly journals Serologic Response of Horses to the Structural Proteins of Equine Arteritis Virus

1998 ◽  
Vol 10 (3) ◽  
pp. 229-236 ◽  
Author(s):  
N. James MacLachlan ◽  
Udeni B. R. Balasuriya ◽  
Jodi F. Hedges ◽  
Therese M. Schweidler ◽  
William H. McCollum ◽  
...  
Keyword(s):  
Humoral Response ◽  
Equine Arteritis Virus ◽  
Structural Proteins ◽  
Structural Protein ◽  
Serum Samples ◽  
Equine Viral Arteritis ◽  
Serologic Response ◽  
Immunoblotting Assay ◽  
Duration Of Infection ◽  
Standard Serum

Equine arteritis virus (EAV) is the causative agent of equine viral arteritis, an apparently emerging disease of equids. In this study, the antibody response of horses to the structural proteins of EAV was evaluated using gradient-purified EAV virions and baculovirus-expressed recombinant EAV structural proteins (GL, GS, M, N) as antigens in a Western immunoblotting assay. Thirty-three sera from horses that previously had been naturally or experimentally infected with EAV were evaluated, including samples from mares, geldings, and both persistently and nonpersistently infected stallions. Sera also were evaluated from 4 horses that had been vaccinated with the commercial modified live EAV vaccine. The data suggest that the serologic response of individual horses to EAV may vary with the infecting virus strain and duration of infection. The M protein was most consistently recognized by the various serum samples, whereas the response to the N and GL proteins was variable and the GS protein was bound by only 1 serum sample. The immunoblotting assay definitively established the protein specificity of the humoral response of horses to EAV; however, it clearly is less sensitive than the standard serum neutralization (SN) test—2 of the 37 sera that were serpositive by th SN test failed to react in the immunoblot assay with any EAV structural protein. Furthermore, the GL protein expresses the known neutralization determinants of EAV, yet only 22 of the 37 sera that had SN antibodies bound the GL protein in the immunoblotting assay. Information from this study will assist ongoing efforts to develop improved methods for the serologic diagnosis of EAV infection of horses.

Monoclonal antibody defining a human syncytiotrophoblastic polypeptide immunologically related to mammalian retrovirus structural protein p30

Placenta ◽  
1984 ◽  
Vol 5 (6) ◽  
pp. 465-473 ◽  
Author(s):  
T. Wahlstroöm ◽  
P. Nieminen ◽  
A. Närvänen ◽  
J. Suni ◽  
P. Lehtovirta ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Structural Protein

scholarly journals Genetic Manipulation of Arterivirus Alternative mRNA Leader-Body Junction Sites Reveals Tight Regulation of Structural Protein Expression

2000 ◽  
Vol 74 (24) ◽  
pp. 11642-11653 ◽  
Author(s):  
Alexander O. Pasternak ◽  
Alexander P. Gultyaev ◽  
Willy J. M. Spaan ◽  
Eric J. Snijder
Keyword(s):  
Rna Structure ◽  
The Body ◽  
Equine Arteritis Virus ◽  
Site Directed Mutagenesis ◽  
Structural Proteins ◽  
Limiting Factors ◽  
Progeny Virus ◽  
Structural Protein ◽  
Sequence Element ◽  
Conserved Sequence

ABSTRACT To express its structural proteins, the arterivirus Equine arteritis virus (EAV) produces a nested set of six subgenomic (sg) RNA species. These RNA molecules are generated by a mechanism of discontinuous transcription, during which a common leader sequence, representing the 5′ end of the genomic RNA, is attached to the bodies of the sg RNAs. The connection between the leader and body parts of an mRNA is formed by a short, conserved sequence element termed the transcription-regulating sequence (TRS), which is present at the 3′ end of the leader as well as upstream of each of the structural protein genes. With the exception of RNA3, only one body TRS was previously assumed to be used to join the leader and body of each EAV sg RNA. Here we show that for the synthesis of two other sg RNAs, RNA4 and RNA5, alternative leader-body junction sites that differ substantially in transcriptional activity are used. By site-directed mutagenesis of an EAV infectious cDNA clone, the alternative TRSs used to generate RNA3, -4, and -5 were inactivated, which strongly influenced the corresponding RNA levels and the production of infectious progeny virus. The relative amounts of RNA produced from alternative TRSs differed significantly and corresponded to the relative infectivities of the virus mutants. This strongly suggested that the structural proteins that are expressed from these RNAs are limiting factors during the viral life cycle and that the discontinuous step in sg RNA synthesis is crucial for the regulation of their expression. On the basis of a theoretical analysis of the predicted RNA structure of the 3′ end of the EAV genome, we propose that the local secondary RNA structure of the body TRS regions is an important factor in the regulation of the discontinuous step in EAV sg mRNA synthesis.

scholarly journals Generation and characterization of a monoclonal antibody against duck Tembusu virus envelope protein

2016 ◽  
Vol 19 (4) ◽  
pp. 877-883 ◽  
Author(s):  
K. Han ◽  
D. Zhao ◽  
Y. Liu ◽  
Q. Liu ◽  
X. Huang ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Elisa Test ◽  
Antibody Detection ◽  
Economic Losses ◽  
Structural Protein ◽  
Neutralization Titer ◽  
Viral Attachment ◽  
E Protein ◽  
Duck Tembusu Virus ◽  
Tembusu Virus

Abstract Duck Tembusu virus (DTMUV) is a newly emerging pathogenic flavivirus that has caused massive economic losses to the duck industry in China. Envelope (E) protein of DTMUV is an important structural protein, which is able to induce protective immune response in target animals and can be used as specific serological diagnosis tool. In this study, a novel monoclonal antibody, designated mAb 3E9, was generated against DTMUV E protein. It is positive in indirect ELISA against both His-E protein and the purified whole viral antigen. Also, this mAb showed positive reaction with DTMUV in Western blot and indirect immunofluorescence assay, and the isotype was IgG1. End-point neutralizing assay performed in BHK-21 cells revealed that the neutralization titer of 3E9 against DTMUV JS804 strain reached 1:50. Furthermore, functional studies revealed that 3E9 blocks infection of DTMUV at a step on viral attachment. The anti-E mAbs produced in the present work may be valuable in developing an antigen-capture ELISA test for antigen detection or a competitive ELISA test for antibody detection or therapeutic medicine for DTMUV in poultry.

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