scholarly journals Two-Helper RNA System for Production of Recombinant Semliki Forest Virus Particles

1999 ◽  
Vol 73 (2) ◽  
pp. 1092-1098 ◽  
Author(s):  
C. Smerdou ◽  
P. Liljeström
Keyword(s):  
Semliki Forest Virus ◽  
Foot And Mouth Disease ◽  
Heterologous Protein Expression ◽  
Lacz Gene ◽  
Rna Recombination ◽  
Rna Molecules ◽  
Virus Particles ◽  
Translational Enhancer ◽  
Mouth Disease Virus ◽  
Spike Proteins

ABSTRACT Alphavirus expression systems based on suicidal virus particles carrying recombinant replicons have proven to be a very efficient way to deliver genes for heterologous protein expression. However, present strategies for production of such particles have biosafety limitations due to the generation, by RNA recombination, of replication-proficient viruses (RPVs). Here we describe a new packaging system for Semliki Forest virus (SFV) based on a the use of a two-helper system in which the capsid and spike proteins of the C-p62-6K-E1 polyprotein are expressed from two independent RNA molecules. The capsid gene contains a translational enhancer and therefore that sequence was also engineered in front of the spike sequence p62-6K-E1. A sequence coding for the foot-and-mouth disease virus 2A autoprotease was inserted in frame between the capsid translational enhancer and the spike genes. This allows production of the spike proteins at high levels with cotranslational removal of the enhancer sequence and normal biosynthesis of the spike complex. The autoprotease activity of the capsid protein was abolished by mutation, further increasing the biosafety of the system. Cotransfection of cells with both helper RNAs and an SFV vector replicon carrying the LacZ gene led to production of recombinant particles with titers of up to 8 × 108 particles per 106 cells. Extensive analysis failed to demonstrate the presence of any RPVs, emphasizing the high biosafety of the system based on two-helper RNAs.

Foot-and-mouth disease virus particles inactivated with binary ethylenimine are efficiently internalized into cultured cells

Vaccine ◽  
2011 ◽  
Vol 29 (52) ◽  
pp. 9655-9662 ◽  
Author(s):  
Miguel A. Martín-Acebes ◽  
Ángela Vázquez-Calvo ◽  
Mónica González-Magaldi ◽  
Francisco Sobrino
Keyword(s):  
Disease Virus ◽  
Cultured Cells ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Virus Particles ◽  
Mouth Disease Virus ◽  
Foot And Mouth

scholarly journals Modification of picornavirus genomic RNA using ‘click’ chemistry shows that unlinking of the VPg peptide is dispensable for translation and replication of the incoming viral RNA

2013 ◽  
Vol 42 (4) ◽  
pp. 2473-2482 ◽  
Author(s):  
Martijn A. Langereis ◽  
Qian Feng ◽  
Frank H. T. Nelissen ◽  
Richard Virgen-Slane ◽  
Gerbrand J. van der Heden van Noort ◽  
...  
Keyword(s):  
Click Reaction ◽  
Enterovirus 71 ◽  
Foot And Mouth Disease ◽  
Viral Rna ◽  
Host Protein ◽  
Genomic Rna ◽  
Rna Molecules ◽  
Rna Translation ◽  
Mouth Disease Virus ◽  
Covalently Linked

Abstract Picornaviruses constitute a large group of viruses comprising medically and economically important pathogens such as poliovirus, coxsackievirus, rhinovirus, enterovirus 71 and foot-and-mouth disease virus. A unique characteristic of these viruses is the use of a viral peptide (VPg) as primer for viral RNA synthesis. As a consequence, all newly formed viral RNA molecules possess a covalently linked VPg peptide. It is known that VPg is enzymatically released from the incoming viral RNA by a host protein, called TDP2, but it is still unclear whether the release of VPg is necessary to initiate RNA translation. To study the possible requirement of VPg release for RNA translation, we developed a novel method to modify the genomic viral RNA with VPg linked via a ‘non-cleavable’ bond. We coupled an azide-modified VPg peptide to an RNA primer harboring a cyclooctyne [bicyclo[6.1.0]nonyne (BCN)] by a copper-free ‘click’ reaction, leading to a VPg-triazole-RNA construct that was ‘non-cleavable’ by TDP2. We successfully ligated the VPg-RNA complex to the viral genomic RNA, directed by base pairing. We show that the lack of VPg unlinkase does not influence RNA translation or replication. Thus, the release of the VPg from the incoming viral RNA is not a prerequisite for RNA translation or replication.

Research on Measures of Reducing Environmental Pollutions by Empty Capsid Virus Particles with Vaccine of Foot and Mouth Disease Virus to Control FMD

2014 ◽  
Vol 1023 ◽  
pp. 257-261
Author(s):  
Qi Hong ◽  
Zi Hong Liu ◽  
Duan Dan Han
Keyword(s):  
Virus Particle ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Promising Candidate ◽  
Effective Measure ◽  
Virus Particles ◽  
Empty Capsid ◽  
Mouth Disease Virus ◽  
Foot And Mouth ◽  
Potential Benefits

Foot-and-mouth disease (FMD) is a highly contagious disease in cloven-hoofed animals, and had been broken out worldwide several times in recent years. In order to extinct outbreak of FMD, a large number of infected animals are slaughtered in some countries, and the slaughterings had been caused a series of environmental pollutions seriously. To reduce this kind of pollution, vaccination is an effective measure to protect animals against FMD, however, FMD virus (FMDV) escaping from manufacturing plant and inactivated incompletely during vaccine production could cause an outbreak of FMD. Therefore, inactivated FMDV vaccines are not safe to animals and environment. FMDV empty capsid (lacking nucleic acid) can elicit the same antibody response as infectious FMDV, thus, empty capsid virus particle vaccine of FMDV would be the most promising candidate vaccine for its safety and protection against FMDV. In this report, we studied the empty capsid virus particle vaccine of FMDV to control FMD and its potential benefits to the environment.

scholarly journals Sequence adaptations affecting cleavage of the VP1/2A junction by the 3C protease in foot-and-mouth disease virus-infected cells

2014 ◽  
Vol 95 (11) ◽  
pp. 2402-2410 ◽  
Author(s):  
Maria Gullberg ◽  
Charlotta Polacek ◽  
Graham J. Belsham
Keyword(s):  
Disease Virus ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Single Amino Acid ◽  
Virus Particles ◽  
3C Protease ◽  
Serotype O ◽  
Single Amino Acid Residue ◽  
Mouth Disease Virus ◽  
Foot And Mouth

The foot-and-mouth disease virus (FMDV) capsid protein precursor P1-2A is cleaved by the virus-encoded 3C protease to VP0, VP3, VP1 and 2A. It was shown previously that modification of a single amino acid residue (K210E) within the VP1 protein and close to the VP1/2A cleavage site, inhibited cleavage of this junction and produced ‘self-tagged’ virus particles. A second site substitution (E83K) within VP1 was also observed within the rescued virus [Gullberg et al. (2013). J Virol 87, 11591–11603]. It was shown here that introduction of this E83K change alone into a serotype O virus resulted in the rapid accumulation of a second site substitution within the 2A sequence (L2P), which also blocked VP1/2A cleavage. This suggests a linkage between the E83K change in VP1 and cleavage of the VP1/2A junction. Cells infected with viruses containing the VP1 K210E or the 2A L2P substitutions contained the uncleaved VP1-2A protein. The 2A L2P substitution resulted in the VP1/2A junction being highly resistant to cleavage by the 3C protease, hence it may be a preferred route for ‘tagging’ virus particles.

scholarly journals Hand-foot-and-mouth disease virus receptor KREMEN1 binds the canyon of Coxsackie Virus A10

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Yuguang Zhao ◽  
Daming Zhou ◽  
Tao Ni ◽  
Dimple Karia ◽  
Abhay Kotecha ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Coxsackie Virus ◽  
Virus Surface ◽  
Virus Particles ◽  
Mouth Disease Virus ◽  
Foot And Mouth ◽  
Severe Infections ◽  
Entry Receptor

AbstractCoxsackievirus A10 (CV-A10) is responsible for an escalating number of severe infections in children, but no prophylactics or therapeutics are currently available. KREMEN1 (KRM1) is the entry receptor for the largest receptor-group of hand-foot-and-mouth disease causing viruses, which includes CV-A10. We report here structures of CV-A10 mature virus alone and in complex with KRM1 as well as of the CV-A10 A-particle. The receptor spans the viral canyon with a large footprint on the virus surface. The footprint has some overlap with that seen for the neonatal Fc receptor complexed with enterovirus E6 but is larger and distinct from that of another enterovirus receptor SCARB2. Reduced occupancy of a particle-stabilising pocket factor in the complexed virus and the presence of both unbound and expanded virus particles suggests receptor binding initiates a cascade of conformational changes that produces expanded particles primed for viral uncoating.

Epitopes on foot-and-mouth disease virus particles I. Topology

Virology ◽  
1987 ◽  
Vol 157 (2) ◽  
pp. 516-525 ◽  
Author(s):  
K.C. McCullough ◽  
J.R. Crowther ◽  
W.C. Carpenter ◽  
E. Brocchi ◽  
L. Capucci ◽  
...  
Keyword(s):  
Disease Virus ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Virus Particles ◽  
Mouth Disease Virus ◽  
Foot And Mouth
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