scholarly journals African Swine Fever Virus Ubiquitin-Conjugating Enzyme Interacts With Host Translation Machinery to Regulate the Host Protein Synthesis

2020 ◽  
Vol 11 ◽  
Author(s):  
Lucía Barrado-Gil ◽  
Ana Del Puerto ◽  
Raquel Muñoz-Moreno ◽  
Inmaculada Galindo ◽  
Miguel Ángel Cuesta-Geijo ◽  
...  
Keyword(s):  
Catalytic Domain ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Emerging Diseases ◽  
Fever Virus ◽  
Host Protein ◽  
Mass Spectrometry Analysis ◽  
Initiation Factor ◽  
Translation Machinery ◽  
Conjugating Enzyme

African Swine Fever virus (ASFV) causes one of the most relevant emerging diseases affecting swine, now extended through three continents. The virus has a large coding capacity to deploy an arsenal of molecules antagonizing the host functions. In the present work, we have studied the only known E2 viral-conjugating enzyme, UBCv1 that is encoded by the I215L gene of ASFV. UBCv1 was expressed as an early expression protein that accumulates throughout the course of infection. This versatile protein, bound several types of polyubiquitin chains and its catalytic domain was required for enzymatic activity. High throughput mass spectrometry analysis in combination with a screening of an alveolar macrophage library was used to identify and characterize novel UBCv1-host interactors. The analysis revealed interaction with the 40S ribosomal protein RPS23, the cap-dependent translation machinery initiation factor eIF4E, and the E3 ubiquitin ligase Cullin 4B. Our data show that during ASFV infection, UBCv1 was able to bind to eIF4E, independent from the cap-dependent complex. Our results provide novel insights into the function of the viral UBCv1 in hijacking cellular components that impact the mTORC signaling pathway, the regulation of the host translation machinery, and the cellular protein expression during the ASFV lifecycle.

The L83L ORF of African swine fever virus strain Georgia encodes for a non-essential gene that interacts with the host protein IL-1β

2018 ◽  
Vol 249 ◽  
pp. 116-123 ◽  
Author(s):  
Manuel V. Borca ◽  
Vivian O’Donnell ◽  
Lauren G. Holinka ◽  
Elizabeth Ramírez-Medina ◽  
Benjamin A. Clark ◽  
...  
Keyword(s):  
Virus Strain ◽  
Essential Gene ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Host Protein

scholarly journals The MyD116 African Swine Fever Virus Homologue Interacts with the Catalytic Subunit of Protein Phosphatase 1 and Activates Its Phosphatase Activity

2007 ◽  
Vol 81 (6) ◽  
pp. 2923-2929 ◽  
Author(s):  
José Rivera ◽  
Charles Abrams ◽  
Bruno Hernáez ◽  
Alberto Alcázar ◽  
José M. Escribano ◽  
...  
Keyword(s):  
Protein Phosphatase ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Catalytic Subunit ◽  
Protein Phosphatase 1 ◽  
Initiation Factor ◽  
Wild Type ◽  
Α Subunit ◽  
Initiation Factor 2

ABSTRACT The DP71L protein of African swine fever virus (ASFV) shares sequence similarity with the herpes simplex virus ICP34.5 protein over a C-terminal domain. We showed that the catalytic subunit of protein phosphatase 1 (PP1) interacts specifically with the ASFV DP71L protein in a yeast two-hybrid screen. The chimeric full-length DP71L protein, from ASFV strain Badajoz 71 (BA71V), fused to glutathione S-transferase (DP71L-GST) was expressed in Escherichia coli and shown to bind specifically to the PP1-α catalytic subunit expressed as a histidine fusion protein (6×His-PP1α) in E. coli. The functional effects of this interaction were investigated by measuring the levels of PP1 and PP2A in ASFV-infected Vero cells. This showed that infection with wild-type ASFV strain BA71V activated PP1 between two- and threefold over that of mock-infected cells. This activation did not occur in cells infected with the BA71V isolate in which the DP71L gene had been deleted, suggesting that expression of DP71L leads to PP1 activation. In contrast, no effect was observed on the activity of PP2A following ASFV infection. We showed that infection of cells with wild-type BA71V virus resulted in decreased phosphorylation of the α subunit of eukaryotic initiation factor 2 (eIF-2α). ICP34.5 recruits PP1 to dephosphorylate the α subunit of eukaryotic translational initiation factor 2 (also known as eIF-2α); possibly the ASFV DP71L protein has a similar function.

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 A novel function of African Swine Fever Virus pE66L in inhibition of host translation by the PKR/eIF2α pathway

2020 ◽  
Author(s):  
Zhou Shen ◽  
Chen Chen ◽  
Yilin Yang ◽  
Zhenhua Xie ◽  
Qingying Ao ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Synthesis ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Inhibitory Effect ◽  
Fever Virus ◽  
Host Protein ◽  
Renilla Luciferase ◽  
Host Gene Expression ◽  
Tm Domain

African swine fever virus (ASFV) is one of the most contagious and lethal viruses infecting pigs. This virus is endemic in many countries and has very recently spread to China, but no licensed vaccines or treatments are currently available. Despite extensive research, the basic question of how ASFV-encoded proteins inhibit host translation remains. Here, we examined how ASFV interfered with host translation and optimized viral gene expression. We found that 14 ASFV proteins inhibited Renilla luciferase (Rluc) activity greater than 5-fold, and the protein with the strongest inhibitory effect was pE66L, which was not previously reported. Combined with bioinformatical analysis and biochemical experiment, we determined that the transmembrane (TM) domain (amino acids 13–34) of pE66L was required for the inhibition of host gene expression. Notably, we constructed a recombinant plasmid with the TM domain linked to enhanced green fluorescent protein (EGFP) and further demonstrated that this domain broadly inhibited protein synthesis. Confocal and biochemical analyses indicated that the TM domain might help proteins locate to the endoplasmic reticulum (ER) to suppress translation though the PKR/eIF2α pathway. Deletion of the E66L gene had little effect on virus replication in macrophages, but significantly recovered host gene expression. Taken together, our findings complement studies on the host translation of ASFV proteins and suggest that ASFV pE66L induces host translation shutoff, which is dependent on activation of the PKR/eIF2α pathway. Importance African swine fever virus (ASFV) is a member of the nucleocytoplasmic large DNA virus superfamily that predominantly replicates in the cytoplasm of infected cells. The ASFV double-stranded DNA genome varies in length from approximately 170 to 193 kbp depending on the isolate and contains between 150 and 167 open reading frames (ORFs), of which half the encoded proteins have not been explored. Our study showed that 14 proteins had an obvious inhibitory effect on Renilla luciferase (Rluc) gene synthesis, with pE66L showing the most significant effect. Furthermore, the transmembrane (TM) domain of pE66L broadly inhibited host protein synthesis in a PKR/eIF2a pathway-dependent manner. Loss of pE66L during ASFV infection had little effect on virus replication, but significantly recovered host protein synthetic. Based on the above results, our findings expand our view of ASFV in determining the fate of host-pathogen interactions.

scholarly journals A ubiquitin conjugating enzyme encoded by African swine fever virus.

1992 ◽  
Vol 11 (1) ◽  
pp. 361-366 ◽  
Author(s):  
P.M. Hingamp ◽  
J.E. Arnold ◽  
R.J. Mayer ◽  
L.K. Dixon
Keyword(s):  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Ubiquitin Conjugating Enzyme ◽  
Conjugating Enzyme

The Ep152R ORF of African swine fever virus strain Georgia encodes for an essential gene that interacts with host protein BAG6

2016 ◽  
Vol 223 ◽  
pp. 181-189 ◽  
Author(s):  
Manuel V. Borca ◽  
Vivian O’Donnell ◽  
Lauren G. Holinka ◽  
Devendra K. Rai ◽  
Brenton Sanford ◽  
...  
Keyword(s):  
Virus Strain ◽  
Essential Gene ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Host Protein

scholarly journals Construction, Identification and Analysis of the Interaction Network of African Swine Fever Virus MGF360-9L with Host Proteins

Viruses ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 1804
Author(s):  
Bo Yang ◽  
Dajun Zhang ◽  
Xijuan Shi ◽  
Chaochao Shen ◽  
Yu Hao ◽  
...  
Keyword(s):  
Protein Interaction ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Interaction Network ◽  
Fever Virus ◽  
Host Protein ◽  
Cellular Proteins ◽  
Regulatory Subunit ◽  
Transcriptional Level

African swine fever virus (ASFV) is prevalent in many countries and is a contagious and lethal virus that infects pigs, posing a threat to the global pig industry and public health. The interaction between the virus and the host is key to unlocking the mystery behind viral pathogenesis. A comprehensive understanding of the viral and host protein interaction may provide clues for developing new antiviral strategies. Here, we show a network of ASFV MGF360-9L protein interactions in porcine kidney (PK-15) cells. Overall, 268 proteins that interact with MGF360-9L are identified using immunoprecipitation and liquid chromatography–mass spectrometry (LC-MS). Accordingly, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted, and the protein–protein interaction (PPI) network was created. It was speculated that the cellular proteins interacting with MGF360-9L are involved in protein binding, metabolism, and the innate immune response. Proteasome subunit alpha type (PSMA3), 26S protease regulatory subunit 4 (PSMC1), autophagy and beclin 1 regulator 1 (AMBRA1), and DEAD-box helicase 20 (DDX20) could interact with MGF360-9L in vitro. PSMA3 and PSMC1 overexpression significantly promoted ASFV replication, and MGF360-9L maintained the transcriptional level of PSMA3 and PSMC1. Here, we show the interaction between ASFV MGF360-9L and cellular proteins and elucidate the virus–host interaction network, which effectively provides useful protein-related information that can enable further study of the potential mechanism and pathogenesis of ASFV infection.

Sign in / Sign up

Export Citation Format

Share Document