scholarly journals The pH Stability of Foot-and-Mouth Disease Virus Particles Is Modulated by Residues Located at the Pentameric Interface and in the N Terminus of VP1

2015 ◽  
Vol 89 (10) ◽  
pp. 5633-5642 ◽  
Author(s):  
Flavia Caridi ◽  
Angela Vázquez-Calvo ◽  
Francisco Sobrino ◽  
Miguel A. Martín-Acebes
Keyword(s):  
Disease Virus ◽  
Molecular Mechanisms ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Acid Sensitivity ◽  
Mouth Disease Virus ◽  
N Terminus ◽  
Foot And Mouth ◽  
Capsid Stability ◽  
Acid Labile

ABSTRACTThe picornavirus foot-and-mouth disease virus (FMDV) is the etiological agent of a highly contagious disease that affects important livestock species. The FMDV capsid is highly acid labile, and viral particles lose infectivity due to their disassembly at pH values slightly below neutrality. This acid sensitivity is related to the mechanism of viral uncoating and genome penetration from endosomes. In this study, we have analyzed the molecular basis of FMDV acid-induced disassembly by isolating and characterizing a panel of novel FMDV mutants differing in acid sensitivity. Amino acid replacements altering virion stability were preferentially distributed in two different regions of the capsid: the N terminus of VP1 and the pentameric interface. Even more, the acid labile phenotype induced by a mutation located at the pentameric interface in VP3 could be compensated by introduction of an amino acid substitution in the N terminus of VP1. These results indicate that the acid sensitivity of FMDV can be considered a multifactorial trait and that virion stability is the fine-tuned product of the interaction between residues from different capsid proteins, in particular those located within the N terminus of VP1 or close to the pentameric interface.IMPORTANCEThe viral capsid protects the viral genome from environmental factors and contributes to virus dissemination and infection. Thus, understanding of the molecular mechanisms that modulate capsid stability is of interest for the basic knowledge of the biology of viruses and as a tool to improve the stability of conventional vaccines based on inactivated virions or empty capsids. Using foot-and-mouth disease virus (FMDV), which displays a capsid with extreme acid sensitivity, we have performed a genetic study to identify the molecular determinants involved in capsid stability. A panel of FMDV mutants with differential sensitivity to acidic pH was generated and characterized, and the results showed that two different regions of FMDV capsid contribute to modulating viral particle stability. These results provide new insights into the molecular mechanisms of acid-mediated FMDV uncoating.

scholarly journals Generation of Antibodies against Foot-and-Mouth-Disease Virus Capsid Protein VP4 Using Hepatitis B Core VLPs as a Scaffold

Life ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 338
Author(s):  
Jessica Swanson ◽  
Rennos Fragkoudis ◽  
Philippa C. Hawes ◽  
Joseph Newman ◽  
Alison Burman ◽  
...  
Keyword(s):  
Amino Acids ◽  
Hepatitis B ◽  
Capsid Protein ◽  
Disease Virus ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Specific Antibodies ◽  
Mouth Disease Virus ◽  
N Terminus ◽  
Foot And Mouth

The picornavirus foot-and-mouth disease virus (FMDV) is the causative agent of the economically important disease of livestock, foot-and-mouth disease (FMD). VP4 is a highly conserved capsid protein, which is important during virus entry. Previous published work has shown that antibodies targeting the N-terminus of VP4 of the picornavirus human rhinovirus are broadly neutralising. In addition, previous studies showed that immunisation with the N-terminal 20 amino acids of enterovirus A71 VP4 displayed on the hepatitis B core (HBc) virus-like particles (VLP) can induce cross-genotype neutralisation. To investigate if a similar neutralising response against FMDV VP4 could be generated, HBc VLPs displaying the N-terminus of FMDV VP4 were designed. The N-terminal 15 amino acids of FMDV VP4 was inserted into the major immunodominant region. HBc VLPs were also decorated with peptides of the N-terminus of FMDV VP4 attached using a HBc-spike binding tag. Both types of VLPs were used to immunise mice and the resulting serum was investigated for VP4-specific antibodies. The VLP with VP4 inserted into the spike, induced VP4-specific antibodies, however the VLPs with peptides attached to the spikes did not. The VP4-specific antibodies could recognise native FMDV, but virus neutralisation was not demonstrated. This work shows that the HBc VLP presents a useful tool for the presentation of FMDV capsid epitopes.

scholarly journals An Engineered Maturation Cleavage Provides a Recombinant Mimic of Foot-and-Mouth Disease Virus Capsid Assembly-Disassembly

Life ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 500
Author(s):  
Joseph Newman ◽  
David J. Rowlands ◽  
Tobias J. Tuthill
Keyword(s):  
Capsid Protein ◽  
Disease Virus ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Virion Assembly ◽  
Capsid Precursor ◽  
Mouth Disease Virus ◽  
Foot And Mouth ◽  
Assembly Intermediate ◽  
Capsid Stability

Picornavirus capsids are assembled from 60 copies of a capsid precursor via a pentameric assembly intermediate or ‘pentamer’. Upon completion of virion assembly, a maturation event induces a final cleavage of the capsid precursor to create the capsid protein VP4, which is essential for capsid stability and entry into new cells. For the picornavirus foot-and-mouth disease virus (FMDV), intact capsids are temperature and acid-labile and can disassemble into pentamers. During disassembly, capsid protein VP4 is lost, presumably altering the structure and properties of the resulting pentamers. The purpose of this study was to compare the characteristics of recombinant “assembly” and “disassembly” pentamers. We generated recombinant versions of these different pentamers containing an engineered cleavage site to mimic the maturation cleavage. We compared the sedimentation and antigenic characteristics of these pentamers using sucrose density gradients and reactivity with an antibody panel. Pentamers mimicking the assembly pathway sedimented faster than those on the disassembly pathway suggesting that for FMDV, in common with other picornaviruses, assembly pentamers sediment at 14S whereas only pentamers on the disassembly pathway sediment at 12S. The reactivity with anti-VP4 antibodies was reduced for the 12S pentamers, consistent with the predicted loss of VP4. Reactivity with other antibodies was similar for both pentamers suggesting that major antigenic features may be preserved between the VP4 containing assembly pentamers and the disassembly pentamers lacking VP4.

scholarly journals Molecular Mechanisms of Immune Escape for Foot-and-Mouth Disease Virus

Pathogens ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 729
Author(s):  
Bo Yang ◽  
Xiaohui Zhang ◽  
Dajun Zhang ◽  
Jing Hou ◽  
GuoWei Xu ◽  
...  
Keyword(s):  
Immune Responses ◽  
Disease Virus ◽  
Molecular Mechanisms ◽  
Immune Escape ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Host Immune Responses ◽  
Mouth Disease Virus ◽  
Foot And Mouth ◽  
Vesicular Disease

Foot-and-mouth disease virus (FMDV) causes a highly contagious vesicular disease in cloven-hoofed livestock that results in severe consequences for international trade, posing a great economic threat to agriculture. The FMDV infection antagonizes the host immune responses via different signaling pathways to achieve immune escape. Strategies to escape the cell immune system are key to effective infection and pathogenesis. This review is focused on summarizing the recent advances to understand how the proteins encoded by FMDV antagonize the host innate and adaptive immune responses.

scholarly journals Thermostability of the Foot-and-Mouth Disease Virus Capsid Is Modulated by Lethal and Viability-Restoring Compensatory Amino Acid Substitutions

2019 ◽  
Vol 93 (10) ◽  
Author(s):  
Silvia López-Argüello ◽  
Verónica Rincón ◽  
Alicia Rodríguez-Huete ◽  
Encarnación Martínez-Salas ◽  
Graham J. Belsham ◽  
...  
Keyword(s):  
Amino Acid ◽  
Disease Virus ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Virus Infectivity ◽  
Side Chains ◽  
Mouth Disease Virus ◽  
Foot And Mouth ◽  
Capsid Stability ◽  
The Relationship

ABSTRACTInfection by viruses depends on a balance between capsid stability and dynamics. This study investigated biologically and biotechnologically relevant aspects of the relationship in foot-and-mouth disease virus (FMDV) between capsid structure and thermostability and between thermostability and infectivity. In the FMDV capsid, a substantial number of amino acid side chains at the interfaces between pentameric subunits are charged at neutral pH. Here a mutational analysis revealed that the essential role for virus infection of most of the 8 tested charged groups is not related to substantial changes in capsid protein expression or processing or in capsid assembly or stability against a thermally induced dissociation into pentamers. However, the positively charged side chains of R2018 and H3141, located at the interpentamer interfaces close to the capsid 2-fold symmetry axes, were found to be critical both for virus infectivity and for keeping the capsid in a state of weak thermostability. A charge-restoring substitution (N2019H) that was repeatedly fixed during amplification of viral genomes carrying deleterious mutations reverted both the lethal and capsid-stabilizing effects of the substitution H3141A, leading to a double mutant virus with close to normal infectivity and thermolability. H3141A and other thermostabilizing substitutions had no detectable effect on capsid resistance to acid-induced dissociation into pentamers. The results suggest that FMDV infectivity requires limited local stability around the 2-fold axes at the interpentamer interfaces of the capsid. The implications for the mechanism of genome uncoating in FMDV and the development of thermostabilized vaccines against foot-and-mouth disease are discussed.IMPORTANCEThis study provides novel insights into the little-known structural determinants of the balance between thermal stability and instability in the capsid of foot-and-mouth disease virus and into the relationship between capsid stability and virus infectivity. The results provide new guidelines for the development of thermostabilized empty capsid-based recombinant vaccines against foot-and-mouth disease, one of the economically most important animal diseases worldwide.

scholarly journals CLONING AND EXPRESSION OF Lb-PROTEASE FROM cDNA CLONE OF FOOT-AND- MOUTH DISEASE VIRUS

2017 ◽  
Vol 5 (9(SE)) ◽  
pp. 62-71
Author(s):  
Swaroop Sarkar ◽  
V.V.S. Suryanarayana ◽  
S.R.Madhan Shankar
Keyword(s):  
Disease Virus ◽  
Cdna Clone ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Host Protein ◽  
Cloning And Expression ◽  
Initiation Factor ◽  
Mouth Disease Virus ◽  
N Terminus ◽  
Foot And Mouth

Foot-and –Mouth disease virus (FMDV) is a positive sense RNA virus and the genome codes for single polyprotein. The FMDV L protein is located at the N terminus of the polyprotein and is the first gene product released from the nascent polyprotein. The leader L protease which is involved in pathogenesis has two known functions: (i) auto-catalytic removal from the N terminus of the viral polyprotein and (ii) cleavage of the p220 subunit of the eukaryotic initiation factor 4F complex, which helps to shut off host protein synthesis. To explore the role of L protease in FMDV pathogenesis we generated synthetic FMDV genome lacking the L gene. The gene was amplified from an infectious cDNA clone of serotype Asia1. Primers corresponding to L protease were designed based on the sequence available in the data base. An amplified DNA of 546bp was purified and cloned into pET28 cloning vector. The sequence analysis revealed the presence of single Open Reading Frame (ORF) encoding a protein of 173 amino acid residues. The sequence alignment using BLAST search in NCBI gene Bank showed 91% homology with FMDV strain A isolate IND17/77 L protease gene. The recombinant plasmids pETLb was transferred into BL21 (DE3) pLysS cells and the IPTG induced expressed protein of 25 KDa was purified by nickel affinity column as per the manufacturer’s protocol (Sigma, USA). The specificity of the expressed protein in was confirmed by western blotting using convalescent cattle serum/ rabbit anti-bovine horse radish peroxidase conjugate and O-Dianisidine Dihydrochloride substrate.

scholarly journals Viral RNA modulates the acid sensitivity of foot-and-mouth disease virus capsids.

1995 ◽  
Vol 69 (1) ◽  
pp. 430-438 ◽  
Author(s):  
S Curry ◽  
C C Abrams ◽  
E Fry ◽  
J C Crowther ◽  
G J Belsham ◽  
...  
Keyword(s):  
Disease Virus ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Viral Rna ◽  
Acid Sensitivity ◽  
Mouth Disease Virus ◽  
Virus Capsids ◽  
Foot And Mouth

scholarly journals Genetic Determinants of Virulence between Two Foot-and-Mouth Disease Virus Isolates Which Caused Outbreaks of Differing Severity

mSphere ◽  
2019 ◽  
Vol 4 (4) ◽  
Author(s):  
Tatsuya Nishi ◽  
Kazuki Morioka ◽  
Nobuko Saito ◽  
Makoto Yamakawa ◽  
Toru Kanno ◽  
...  
Keyword(s):  
Disease Virus ◽  
Molecular Mechanisms ◽  
Three Dimensional ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Genetic Determinants ◽  
Replication Fidelity ◽  
Suckling Mice ◽  
Mouth Disease Virus ◽  
Foot And Mouth

ABSTRACT Individual foot-and-mouth disease virus (FMDV) strains reveal different degrees of infectivity and pathogenicity in host animals. The differences in severity among outbreaks might be ascribable to these differences in infectivity among FMDV strains. To investigate the molecular mechanisms underlying these differences, we estimated the infectivity of O/JPN/2000 and O/JPN/2010, which caused outbreaks of markedly different scales, in cell lines, Holstein cattle, and suckling mice. Viral growth of the two strains in cells was not remarkably different; however, O/JPN/2000 showed apparently low transmissibility in cattle. Mortality rates of suckling mice inoculated intraperitoneally with a 50% tissue culture infective dose (TCID50) of 10 for O/JPN/2000 and O/JPN/2010 also differed, at 0% and 100%, respectively. To identify genes responsible for this difference in infectivity, genetic regions of the full-length cDNA of O/JPN/2010 were replaced with corresponding fragments of O/JPN/2000. A total of eight recombinant viruses were successfully recovered, and suckling mice were intraperitoneally inoculated. Strikingly, recombinants having either VP1 or 3D derived from O/JPN/2000 showed 0% mortality in suckling mice, whereas other recombinants showed 100% mortality. This finding indicates that VP1, the outermost component of the virus particle, and 3D, an RNA-dependent RNA polymerase, are individually involved in the virulence of O/JPN/2010. Three-dimensional structural analysis of VP1 confirmed that amino acid differences between the two strains were located mainly at the domain interacting with the cellular receptor. On the other hand, measurement of their mutation frequencies demonstrated that O/JPN/2000 had higher replication fidelity than O/JPN/2010. IMPORTANCE Efforts to understand the universal mechanism of foot-and-mouth disease virus (FMDV) infection may be aided by knowledge of the molecular mechanisms which underlie differences in virulence beyond multiple topotypes and serotypes of FMDV. Here, we demonstrated independent genetic determinants of two FMDV isolates which have different transmissibility in cattle, namely, VP1 and 3D protein. Findings suggested that the selectivity of VP1 for host cell receptors and replication fidelity during replication were important individual factors in the induction of differences in virulence in the host as well as in the severity of outbreaks in the field. These findings will aid the development of safe live vaccines and antivirals which obstruct viral infection in natural hosts.

scholarly journals The cis-responsive element of Foot-and-mouth disease virus interacts with host cellular factor PCBP2 dependent on host specificity

2021 ◽  
Author(s):  
JaeHun Cheong ◽  
Jeong A Jang ◽  
Bok Kyung Ku
Keyword(s):  
Disease Virus ◽  
Molecular Mechanisms ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Genome Replication ◽  
Viral Pathogen ◽  
Cellular Factor ◽  
Mouth Disease Virus ◽  
Foot And Mouth ◽  
Responsive Element

Abstract I. Background: Foot-and-mouth disease virus (FMDV) is a highly contagious viral pathogen in cloven-hoofed animal including cattle and pig, yet progress in the molecular mechanisms of FMDV genome replication is notably lagging behind that for many RNA viruses. A positive single stranded RNA of FMDV encodes a single long open reading frame flanked by a long 5’-untranslated region (5’UTR) and a short 3’-UTR. The cis-responsive element (CRE) of 5’UTR is critical for FMDV genome replication. II. Methods and Results: Here, we described that poly(C)-binding protein 2 (PCBP2) is revealed as a CRE-binding cellular factor. The RNA immunoprecipitation experiment confirmed that the FMDV CRE interacts with PCBP2 protein. CRE derived from FMDV infection in pig bound stronger to PCBP2 protein of pig than cattle PCBP2, showing host specific RNA-protein interaction. In addition, PCBP2 interacts with FMDV 3B protein together with CRE. The interaction of PCBP and 3B protein with CRE also showed host-specific manners. III. Conclusions: These data suggest that cellular PCBP2 may serve as a host cellular factor of FMDV to facilitate viral replication through interaction with the viral genome and contribute to determine host susceptibility of FMDV variants. The inter-molecular interaction between cellular PCBP2 and FMDV 3B and CRE provides perspectives for antiviral strategy.

High resolution surface structure of foot-and-mouth disease virus

1978 ◽  
Vol 36 (2) ◽  
pp. 376-377
Author(s):  
S. S. Breese ◽  
H. L. Bachrach
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Surface Structure ◽  
Disease Virus ◽  
Potassium Phosphate ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Physical Measurements ◽  
Mouth Disease Virus ◽  
Foot And Mouth

Models for the structure of foot-and-mouth disease virus (FMDV) have been proposed from chemical and physical measurements (Brown, et al., 1970; Talbot and Brown, 1972; Strohmaier and Adam, 1976) and from rotational image-enhancement electron microscopy (Breese, et al., 1965). In this report we examine the surface structure of FMDV particles by high resolution electron microscopy and compare it with that of particles in which the outermost capsid protein VP3 (ca. 30, 000 daltons) has been split into smaller segments, two of which VP3a and VP3b have molecular weights of about 15, 000 daltons (Bachrach, et al., 1975).Highly purified and concentrated type A12, strain 119 FMDV (5 mg/ml) was prepared as previously described (Bachrach, et al., 1964) and stored at 4°C in 0. 2 M KC1-0. 5 M potassium phosphate buffer at pH 7. 5. For electron microscopy, 1. 0 ml samples of purified virus and trypsin-treated virus were dialyzed at 4°C against 0. 2 M NH4OAC at pH 7. 3, deposited onto carbonized formvar-coated copper screens and stained with phosphotungstic acid, pH 7. 3.

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