scholarly journals Evaluation of the Function of the ASFV KP177R Gene, Encoding for Structural Protein p22, in the Process of Virus Replication and in Swine Virulence

Viruses ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 986
Author(s):  
Elizabeth A. Vuono ◽  
Elizabeth Ramirez-Medina ◽  
Sarah Pruitt ◽  
Ayushi Rai ◽  
Nallely Espinoza ◽  
...  
Keyword(s):  
Virus Replication ◽  
African Swine Fever Virus ◽  
Field Isolate ◽  
African Swine Fever ◽  
Virus Protein ◽  
Structural Protein ◽  
Host Proteins ◽  
Gene Encoding ◽  
Exact Function

African swine fever virus (ASFV) causes a devastating disease of swine that has caused outbreaks in Central Europe since 2007, spreading into Asia in 2018. ASFV is a large, structurally complex virus with a large dsDNA genome encoding for more than 160 genes, most of them still uncharacterized. p22, encoded by the ASFV gene KP177R, is an early transcribed, structural virus protein located in the ASFV particle. Although its exact function is unknown, p22 has recently been identified as an interacting partner of several host proteins. Here, we describe the development of a recombinant ASFV (ASFV-G-∆KP177R) lacking the KP177R gene as a tool to evaluate the role of p22 in virus replication and virulence in swine. The recombinant ASFV-G-∆KP177R demonstrated that the KP177R gene is non-essential for ASFV replication in primary swine macrophages, with virus yields similar to those of the parental, highly virulent field isolate Georgia2010 (ASFV-G). In addition, experimental infection of domestic pigs with ASFV-G-∆KP177R produced a clinical disease similar to that caused by the parental ASFV-G. Therefore, and surprisingly, p22 does not seem to be involved in virus replication or virulence in swine.

scholarly journals The MGF360-16R ORF of African Swine Fever Virus Strain Georgia Encodes for a Nonessential Gene That Interacts with Host Proteins SERTAD3 and SDCBP

Viruses ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 60 ◽  
Author(s):  
Elizabeth Ramírez-Medina ◽  
Elizabeth A. Vuono ◽  
Lauro Velazquez-Salinas ◽  
Ediane Silva ◽  
Ayushi Rai ◽  
...  
Keyword(s):  
African Swine Fever Virus ◽  
Field Isolate ◽  
African Swine Fever ◽  
Fever Virus ◽  
Host Protein ◽  
Parental Virus ◽  
Virus Protein ◽  
Host Proteins ◽  
Swine Industry ◽  
Nonessential Gene

African swine fever virus (ASFV) causes a contagious and frequently lethal disease of pigs with significant economic consequences to the swine industry. The ASFV genome encodes for more than 160 genes, but only a few of them have been studied in detail. Here we report the characterization of open reading frame (ORF) MGF360-16R. Kinetic studies of virus RNA transcription demonstrated that the MGF360-16R gene is transcribed as a late virus protein. Analysis of host–protein interactions for the MGF360-16R gene using a yeast two-hybrid screen identified SERTA domain containing 3 (SERTAD3) and syndecan-binding protein (SDCBP) as host protein binding partners. SERTAD3 and SDCBP are both involved in nuclear transcription and SDCBP has been shown to be involved in virus traffic inside the host cell. Interaction between MGF360-16R and SERTAD3 and SDCBP host proteins was confirmed in eukaryotic cells transfected with plasmids expressing MGF360-16R and SERTAD3 or SDCBP fused to fluorescent tags. A recombinant ASFV lacking the MGF360-16R gene (ASFV-G-ΔMGF360-16R) was developed from the highly virulent field isolate Georgia2007 (ASFV-G) and was used to show that MGF360-16R is a nonessential gene. ASFV-G-ΔMGF360-16R had a similar replication ability in primary swine macrophage cell cultures when compared to its parental virus ASFV-G. Experimental infection of domestic pigs showed that ASFV-G-ΔMGF360-16R is as virulent as the parental virus ASFV-G.

scholarly journals Role of African swine fever virus (ASFV) proteins EP153R and EP402R in reducing viral persistence and virulence from attenuated BeninΔDP148R

2021 ◽  
Author(s):  
Vlad Petrovan ◽  
Anusyah Rathakrishnan ◽  
Muneeb Islam ◽  
Lynnette Goatley ◽  
Katy Moffat ◽  
...  
Keyword(s):  
Virus Replication ◽  
Vaccine Development ◽  
African Swine Fever Virus ◽  
Clinical Signs ◽  
African Swine Fever ◽  
Viral Persistence ◽  
Fever Virus ◽  
Post Immunization

The limited knowledge on the role of many of the approximately 170 proteins encoded by African swine fever virus restricts progress towards vaccine development. In this study we investigated the effect of deleting combinations of different genes from a previously attenuated virus, BeninΔDP148R on: virus replication in macrophages, virus persistence and clinical signs post immunization, and induction of protection against challenge. Deletion of either EP402R or EP153R genes individually or in combination from BeninΔDP148R did not reduce virus replication in vitro. However, deletion of EP402R dramatically reduced viral persistence in vivo, whilst maintaining high levels of protection against challenge. The additional deletion of EP153R (BeninΔDP148RΔEP153RΔEP402R) further attenuated the virus and no viremia or clinical signs were observed post immunization. This was associated with decreased protection and detection of moderate levels of challenge virus in blood. Interestingly, the deletion of EP153R alone from BeninΔDP148R did not result in further virus attenuation and a slight increase in virus genome copies in blood was observed at different times post immunization when compared with BeninΔDP148R. These results show that EP402R and EP153R have a synergistic role in promoting viremia, however EP153R alone does not seem to have a major impact on virus levels in blood.

scholarly journals Evaluation of an ASFV RNA Helicase Gene A859L for Virus Replication and Swine Virulence

Viruses ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 10
Author(s):  
Elizabeth Ramirez-Medina ◽  
Elizabeth A. Vuono ◽  
Sarah Pruitt ◽  
Ayushi Rai ◽  
Nallely Espinoza ◽  
...  
Keyword(s):  
Virus Replication ◽  
Recombinant Virus ◽  
Bioinformatics Analysis ◽  
African Swine Fever Virus ◽  
Geographical Area ◽  
African Swine Fever ◽  
Rna Helicase ◽  
Helicase Gene ◽  
Highly Virulent

African swine fever virus (ASFV) is producing a devastating pandemic that, since 2007, has spread to a contiguous geographical area from central Europe to Asia. In July 2021, ASFV was detected in the Dominican Republic, the first report of the disease in the Americas in more than 40 years. ASFV is a large, highly complex virus harboring a large dsDNA genome that encodes for more than 150 genes. The majority of these genes have not been functionally characterized. Bioinformatics analysis predicts that ASFV gene A859L encodes for an RNA helicase, although its function has not yet been experimentally assessed. Here, we evaluated the role of the A859L gene during virus replication in cell cultures and during infection in swine. For that purpose, a recombinant virus (ASFV-G-∆A859L) harboring a deletion of the A859L gene was developed using the highly virulent ASFV Georgia (ASFV-G) isolate as a template. Recombinant ASFV-G-∆A859L replicates in swine macrophage cultures as efficiently as the parental virus ASFV-G, demonstrating that the A859L gene is non-essential for ASFV replication. Experimental infection of domestic pigs demonstrated that ASFV-G-∆A859L replicates as efficiently and induces a clinical disease indistinguishable from that caused by the parental ASFV-G. These studies conclude that the predicted RNA helicase gene A859L is not involved in the processes of virus replication or disease production in swine.

scholarly journals African Swine Fever Virus Protein p17 Is Essential for the Progression of Viral Membrane Precursors toward Icosahedral Intermediates

2010 ◽  
Vol 84 (15) ◽  
pp. 7484-7499 ◽  
Author(s):  
Cristina Suárez ◽  
Javier Gutiérrez-Berzal ◽  
Germán Andrés ◽  
María L. Salas ◽  
Javier M. Rodríguez
Keyword(s):  
Early Stage ◽  
African Swine Fever Virus ◽  
Transmembrane Protein ◽  
African Swine Fever ◽  
Integral Membrane Protein ◽  
Proteolytic Processing ◽  
Fever Virus ◽  
Morphological Evidence ◽  
Virus Protein

ABSTRACT The first morphological evidence of African swine fever virus (ASFV) assembly is the appearance of precursor viral membranes, thought to derive from the endoplasmic reticulum, within the assembly sites. We have shown previously that protein p54, a viral structural integral membrane protein, is essential for the generation of the viral precursor membranes. In this report, we study the role of protein p17, an abundant transmembrane protein localized at the viral internal envelope, in these processes. Using an inducible virus for this protein, we show that p17 is essential for virus viability and that its repression blocks the proteolytic processing of polyproteins pp220 and pp62. Electron microscopy analyses demonstrate that when the infection occurs under restrictive conditions, viral morphogenesis is blocked at an early stage, immediately posterior to the formation of the viral precursor membranes, indicating that protein p17 is required to allow their progression toward icosahedral particles. Thus, the absence of this protein leads to an accumulation of these precursors and to the delocalization of the major components of the capsid and core shell domains. The study of ultrathin serial sections from cells infected with BA71V or the inducible virus under permissive conditions revealed the presence of large helicoidal structures from which immature particles are produced, suggesting that these helicoidal structures represent a previously undetected viral intermediate.

scholarly journals The C962R ORF of African Swine Fever Strain Georgia Is Non-Essential and Not Required for Virulence in Swine

Viruses ◽  
2020 ◽  
Vol 12 (6) ◽  
pp. 676 ◽  
Author(s):  
Elizabeth. Ramirez-Medina ◽  
Elizabeth. A. Vuono ◽  
Ayushi. Rai ◽  
Sarah. Pruitt ◽  
Ediane. Silva ◽  
...  
Keyword(s):  
African Swine Fever Virus ◽  
Field Isolate ◽  
Protein A ◽  
African Swine Fever ◽  
Virus Genome ◽  
Economic Losses ◽  
Virus Protein ◽  
Swine Industry ◽  
Reading Frame

African swine fever virus (ASFV) is the causative agent of the African swine fever (ASF) epizootic currently affecting pigs throughout Eurasia, causing significant economic losses in the swine industry. The virus genome encodes for more than 160 genes, of which only a few have been studied in detail. Here we describe the previously uncharacterized ASFV open reading frame (ORF) C962R, a gene encoding for a putative NTPase. RNA transcription studies using infected swine macrophages demonstrate that the C962R gene is translated as a late virus protein. A recombinant ASFV lacking the C962R gene (ASFV-G-ΔC962R) demonstrates in vivo that the C962R gene is non-essential, since ASFV-G-ΔC962R has similar replication kinetics in primary swine macrophage cell cultures when compared to parental highly virulent field isolate Georgia2007 (ASFV-G). Experimental infection of domestic pigs with ASFV-G-ΔC962R produced a clinical disease similar to that caused by the parental ASFV-G, confirming that deletion of the C962R gene from the ASFV genome does not impact virulence.

scholarly journals Rapid Identification for Serotyping of African Swine Fever Virus Based on the Short Fragment of the EP402R Gene Encoding for CD2-Like Protein

2021 ◽  
Vol 71 (1) ◽  
pp. 98-106
Author(s):  
Tran Ha Thi Thanh ◽  
Truong Anh Duc ◽  
Ly Duc Viet ◽  
Hoang Tuan Van ◽  
Nguyen Chinh Thi ◽  
...  
Keyword(s):  
Potential Role ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Rapid Identification ◽  
Previous Method ◽  
Gene Encoding ◽  
North Central ◽  
Short Fragment ◽  
Coast Of Vietnam

Abstract The first confirmed case of African swine fever (ASF) in Vietnam was officially reported in February 2019. To date, the ASF virus (ASFV) has been detected in all 63 cities/provinces in Vietnam. In order to get a better understanding of the potential role of the EP402R gene in a grouping of ASFV serotypes, thirty ASFV sequences of EP402R genes (accession numbers: MN711757-86) from North Central Coast of Vietnam and 68 well-known references of serotype groups from previous studies were further analyzed. Interestingly, we found that a short fragment of 90 nucleotides was very typical for 8 serological groups of ASFVs. A primer set was designed to amplicon the short fragment of 90 nucleotides using the Primer3 program to establish a simplified method for the serotyping of ASFV. Our results indicated that phylogenetic analysis of the short fragment (90 nucleotides) of the EP402R gene is a very specific and useful method for ASFV serotyping when compared to the previous method using a long fragment (816 nucleotides) of this gene and well-known serotype references based on haemadsorption inhibition (HAI) assay.

scholarly journals Evaluation in Swine of a Recombinant African Swine Fever Virus Lacking the MGF-360-1L Gene

Viruses ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 1193 ◽  
Author(s):  
Elizabeth Ramirez-Medina ◽  
Elizabeth A. Vuono ◽  
Ayushi Rai ◽  
Sarah Pruitt ◽  
Ediane Silva ◽  
...  
Keyword(s):  
African Swine Fever Virus ◽  
Field Isolate ◽  
African Swine Fever ◽  
Kinetic Studies ◽  
Virus Protein ◽  
Swine Production ◽  
Virus Rna ◽  
A Genome ◽  
Highly Virulent ◽  
Production Losses

The African swine fever (ASF) pandemic is currently affecting pigs throughout Eurasia, resulting in significant swine production losses. The causative agent, ASF virus (ASFV), is a large, structurally complex virus with a genome encoding more than 160 genes. The function of most of those genes remains unknown. Here, we presented the previously uncharacterized ASFV gene MGF360-1L, the first gene in the genome. The kinetic studies of virus RNA transcription demonstrated that the MGF360-1L gene was transcribed as a late virus protein. The essentiality of MGF360-1L to virus replication was evaluated by developing a recombinant ASFV lacking the gene (ASFV-G-ΔMGF360-1L). In primary swine macrophage cell cultures, ASFV-G-ΔMGF360-1L showed similar replication kinetics as the parental highly virulent field isolate Georgia2007 (ASFV-G). Domestic pigs experimentally infected with ASFV-G-ΔMGF360-1L presented with a clinical disease indistinguishable from that caused by ASFV-G, demonstrating that MGF360-1L was not involved in virulence in swine, the natural host of ASFV.

scholarly journals Mapping and Sequence of the Gene Encoding Protein pl7, a Major African Swine Fever Virus Structural Protein

Virology ◽  
1995 ◽  
Vol 206 (2) ◽  
pp. 1140-1144 ◽  
Author(s):  
Carmen Simón-Mateo ◽  
Jose Maria P. Freue ◽  
German Andrés ◽  
Carlos López-Otin ◽  
Eladio Viñuela
Keyword(s):  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Structural Protein ◽  
Gene Encoding ◽  
Encoding Protein

scholarly journals The Role of Interleukine-10 and Interferon-γ as Potential Markers of the Evolution of African Swine Fever Virus Infection in Wild Boar

Pathogens ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 757
Author(s):  
Sandra Barroso-Arévalo ◽  
Jose A. Barasona ◽  
Estefanía Cadenas-Fernández ◽  
José M. Sánchez-Vizcaíno
Keyword(s):  
Wild Boar ◽  
Vaccine Candidate ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Interferon Γ ◽  
Fever Virus ◽  
Control Group ◽  
Challenge Virus ◽  
Ifn Γ

African swine fever virus (ASFv) is one of the most challenging pathogens to affect both domestic and wild pigs. The disease has now spread to Europe and Asia, causing great damage to the pig industry. Although no commercial vaccine with which to control the disease is, as yet, available, some potential vaccine candidates have shown good results in terms of protection. However, little is known about the host immune mechanisms underlying that protection, especially in wild boar, which is the main reservoir of the disease in Europe. Here, we study the role played by two cytokines (IL-10 and IFN-γ) in wild boar orally inoculated with the attenuated vaccine candidate Lv17/WB/Rie1 and challenged with a virulent ASFv genotype II isolate. A group of naïve wild boar challenged with the latter isolate was also established as a control group. Our results showed that both cytokines play a key role in protecting the host against the challenge virus. While high levels of IL-10 in serum may trigger an immune system malfunctioning in challenged animals, the provision of stable levels of this cytokine over time may help to control the disease. This, together with high and timely induction of IFN-γ by the vaccine candidate, could help protect animals from fatal outcomes. Further studies should be conducted in order to support these preliminary results and confirm the role of these two cytokines as potential markers of the evolution of ASFV infection.

Sign in / Sign up

Export Citation Format

Share Document