scholarly journals African Swine Fever Virus Structural Protein pE120R Is Essential for Virus Transport from Assembly Sites to Plasma Membrane but Not for Infectivity

2001 ◽  
Vol 75 (15) ◽  
pp. 6758-6768 ◽  
Author(s):  
Germán Andrés ◽  
Ramón Garcı́a-Escudero ◽  
Eladio Viñuela ◽  
Marı́a L. Salas ◽  
Javier M. Rodrı́guez
Keyword(s):  
Plasma Membrane ◽  
African Swine Fever Virus ◽  
Growth Curves ◽  
African Swine Fever ◽  
Fever Virus ◽  
Virus Yield ◽  
Virus Infectivity ◽  
Structural Protein ◽  
Virus Growth ◽  
Extracellular Virus

ABSTRACT This report examines the role of African swine fever virus (ASFV) structural protein pE120R in virus replication. Immunoelectron microscopy revealed that protein pE120R localizes at the surface of the intracellular virions. Consistent with this, coimmunoprecipitation assays showed that protein pE120R binds to the major capsid protein p72. Moreover, it was found that, in cells infected with an ASFV recombinant that inducibly expresses protein p72, the incorporation of pE120R into the virus particle is dependent on p72 expression. Protein pE120R was also studied using an ASFV recombinant in which E120R gene expression is regulated by the Escherichia coli lacrepressor-operator system. In the absence of inducer, pE120R expression was reduced about 100-fold compared to that obtained with the parental virus or the recombinant virus grown under permissive conditions. One-step virus growth curves showed that, under conditions that repress pE120R expression, the titer of intracellular progeny was similar to the total virus yield obtained under permissive conditions, whereas the extracellular virus yield was about 100-fold lower than in control infections. Immunofluorescence and electron microscopy demonstrated that, under restrictive conditions, intracellular mature virions are properly assembled but remain confined to the replication areas. Altogether, these results indicate that pE120R is necessary for virus dissemination but not for virus infectivity. The data also suggest that protein pE120R might be involved in the microtubule-mediated transport of ASFV particles from the viral factories to the plasma membrane.

scholarly journals An African swine fever virus ORF with similarity to C-type lectins is non-essential for growth in swine macrophages in vitro and for virus virulence in domestic swine

1999 ◽  
Vol 80 (10) ◽  
pp. 2693-2697 ◽  
Author(s):  
J. G. Neilan ◽  
M. V. Borca ◽  
Z. Lu ◽  
G. F. Kutish ◽  
S. B. Kleiboeker ◽  
...  
Keyword(s):  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Virus Infectivity ◽  
Mrna Transcription ◽  
Host Cell Interactions ◽  
Lectin Family ◽  
Virus Replication Cycle ◽  
Domestic Swine

An African swine fever virus (ASFV) ORF, 8CR, with similarity to the C-type lectin family of adhesion proteins has been described in the pathogenic isolate Malawi Lil-20/1. The similarity of 8CR to cellular and poxvirus genes associated with cell adhesion, cell recognition and virus infectivity suggested that 8CR may be of significance to ASFV–host cell interactions. Sequence analysis of the 8CR ORF from additional pathogenic ASFV isolates demonstrated conservation among isolates from both pig and tick sources. Northern blot analysis demonstrated 8CR mRNA transcription late in the virus replication cycle. A Malawi Lil-20/1 8CR deletion mutant (Δ8CR) was constructed to analyse 8CR function further. The growth characteristics in vitro of Δ8CR in porcine macrophage cell cultures were identical to those observed for parental virus. In domestic swine, Δ8CR exhibited an unaltered parental Malawi Lil- 20/1 disease and virulence phenotype. Thus, although well conserved among pathogenic ASFV field isolates, 8CR is non-essential for growth in porcine macrophages in vitro and for virus virulence in domestic swine.

scholarly journals African Swine Fever Virus pB119L Protein Is a Flavin Adenine Dinucleotide-Linked Sulfhydryl Oxidase

2006 ◽  
Vol 80 (7) ◽  
pp. 3157-3166 ◽  
Author(s):  
Irene Rodríguez ◽  
Modesto Redrejo-Rodríguez ◽  
Javier M. Rodríguez ◽  
Alí Alejo ◽  
José Salas ◽  
...  
Keyword(s):  
Disulfide Bonds ◽  
Flavin Adenine Dinucleotide ◽  
Virus Assembly ◽  
Nonstructural Protein ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Structural Protein ◽  
Sulfhydryl Oxidase ◽  
Infected Cells

ABSTRACT Protein pB119L of African swine fever virus belongs to the Erv1p/Alrp family of sulfhydryl oxidases and has been described as a late nonstructural protein required for correct virus assembly. To further our knowledge of the function of protein pB119L during the virus life cycle, we have investigated whether this protein possesses sulfhydryl oxidase activity, using a purified recombinant protein. We show that the purified protein contains bound flavin adenine dinucleotide and is capable of catalyzing the formation of disulfide bonds both in a protein substrate and in the small molecule dithiothreitol, the catalytic activity being comparable to that of the Erv1p protein. Furthermore, protein pB119L contains the cysteines of its active-site motif CXXC, predominantly in an oxidized state, and forms noncovalently bound dimers in infected cells. We also show in coimmunoprecipitation experiments that protein pB119L interacts with the viral protein pA151R, which contains a CXXC motif similar to that present in thioredoxins. Protein pA151R, in turn, was found to interact with the viral structural protein pE248R, which contains disulfide bridges and belongs to a class of myristoylated proteins related to vaccinia virus L1R, one of the substrates of the redox pathway encoded by this virus. These results suggest the existence in African swine fever virus of a system for the formation of disulfide bonds constituted at least by proteins pB119L and pA151R and identify protein pE248R as a possible final substrate of this pathway.

scholarly journals Characterization of the African Swine Fever Virus Structural Protein p14.5: A DNA Binding Protein

Virology ◽  
1997 ◽  
Vol 229 (1) ◽  
pp. 201-211 ◽  
Author(s):  
Luisa Martinez-Pomares ◽  
Carmen Simon-Mateo ◽  
Carlos Lopez-Otin ◽  
Eladio Viñuela
Keyword(s):  
Dna Binding ◽  
African Swine Fever Virus ◽  
Binding Protein ◽  
African Swine Fever ◽  
Dna Binding Protein ◽  
Fever Virus ◽  
Structural Protein

Structure basis of non-structural protein pA151R from African Swine Fever Virus

2020 ◽  
Vol 532 (1) ◽  
pp. 108-113
Author(s):  
Jian-Wen Huang ◽  
Du Niu ◽  
Ke Liu ◽  
Qian Wang ◽  
Lixin Ma ◽  
...  
Keyword(s):  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Structural Protein

scholarly journals Crystal structure of the African swine fever virus structural protein p35 reveals its role for core shell assembly

2020 ◽  
Vol 11 (8) ◽  
pp. 600-605
Author(s):  
Guobang Li ◽  
Dan Fu ◽  
Guangshun Zhang ◽  
Dongming Zhao ◽  
Mingyu Li ◽  
...  
Keyword(s):  
Crystal Structure ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Core Shell ◽  
Structural Protein

Efficacy of various methods for African swine fever virus growth in hematopoietic cells

2014 ◽  
pp. 58-63
Author(s):  
N.I. Zakutskii ◽  
◽  
T.G. Shirokova ◽  
N.S. Neverovskaya ◽  
S.G. Yurkov ◽  
...  
Keyword(s):  
African Swine Fever Virus ◽  
Hematopoietic Cells ◽  
African Swine Fever ◽  
Fever Virus ◽  
Virus Growth

scholarly journals Comparison of the genome sequences of non-pathogenic and pathogenic African swine fever virus isolates

2008 ◽  
Vol 89 (2) ◽  
pp. 397-408 ◽  
Author(s):  
David A. G. Chapman ◽  
Vasily Tcherepanov ◽  
Chris Upton ◽  
Linda K. Dixon
Keyword(s):  
Virus Assembly ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Open Reading Frames ◽  
Structural Protein ◽  
Genome Sequences ◽  
Virus Genotype ◽  
Pathogenic Isolate ◽  
Virus Isolates

The genomic coding sequences, apart from the inverted terminal repeats and cross-links, have been determined for two African swine fever virus (ASFV) isolates from the same virus genotype, a non-pathogenic isolate from Portugal, OURT88/3, and a highly pathogenic isolate from West Africa, Benin 97/1. These genome sequences were annotated and compared with that of a tissue culture-adapted isolate, BA71V. The genomes range in length between 170 and 182 kbp and encode between 151 and 157 open reading frames (ORFs). Compared to the Benin 97/1 isolate, the OURT88/3 and BA71V isolates have deletions of 8–10 kbp that encode six copies of the multigene family (MGF) 360 and either one MGF 505/530 copy in the BA71V or two copies in the OURT88/3 isolate. The BA71V isolate has a deletion, close to the right end of the genome, of 3 kbp compared with the other isolates. The five ORFs in this region include an additional copy of an ORF similar to that encoding the p22 virus structural protein. The OURT88/3 isolate has interruptions in ORFs that encode a CD2-like and a C-type lectin protein. Variation between the genomes is observed in the number of copies of five different MGFs. The 109 non-duplicated ORFs conserved in the three genomes encode proteins involved in virus replication, virus assembly and modulation of the host's defences. These results provide information concerning the genetic variability of African swine fever virus isolates that differ in pathogenicity.

scholarly journals Transport of African Swine Fever Virus from Assembly Sites to the Plasma Membrane Is Dependent on Microtubules and Conventional Kinesin

2004 ◽  
Vol 78 (15) ◽  
pp. 7990-8001 ◽  
Author(s):  
Nolwenn Jouvenet ◽  
Paul Monaghan ◽  
Michael Way ◽  
Thomas Wileman
Keyword(s):  
Plasma Membrane ◽  
Immunofluorescence Microscopy ◽  
African Swine Fever Virus ◽  
Large Fraction ◽  
African Swine Fever ◽  
Fever Virus ◽  
Cell Periphery ◽  
Microtubule Organizing Center ◽  
Microtubule Motor ◽  
Conventional Kinesin

ABSTRACT African swine fever virus (ASFV) is a large DNA virus that assembles in perinuclear viral factories located close to the microtubule organizing center. In this study, we have investigated the mechanism by which ASFV reaches the cell surface from the site of assembly. Immunofluorescence microscopy revealed that at 16 h postinfection, mature virions were aligned along microtubules. Furthermore, virus movement to the cell periphery was inhibited when microtubules were depolymerized by nocodazole. In addition, ASFV infection resulted in the increased acetylation of microtubules as well as their protection against depolymerization by nocodazole. Immunofluorescence microscopy showed that conventional kinesin was recruited to virus factories and to a large fraction of virus particles in the cytoplasm. Consistent with a role for conventional kinesin during ASFV egress to the cell periphery, overexpression of the cargo-binding domain of the kinesin light chain severely inhibited the movement of particles to the plasma membrane. Based on our observations, we propose that ASFV is recognized as cargo by conventional kinesin and uses this plus-end microtubule motor to move from perinuclear assembly sites to the plasma membrane.

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