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[PMID]:29244872
[Au] Autor:Barrado-Gil L; Galindo I; Martínez-Alonso D; Viedma S; Alonso C
[Ad] Endereço:Department of Biotechnology, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, INIA, Madrid, Spain.
[Ti] Título:The ubiquitin-proteasome system is required for African swine fever replication.
[So] Source:PLoS One;12(12):e0189741, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Several viruses manipulate the ubiquitin-proteasome system (UPS) to initiate a productive infection. Determined viral proteins are able to change the host's ubiquitin machinery and some viruses even encode their own ubiquitinating or deubiquitinating enzymes. African swine fever virus (ASFV) encodes a gene homologous to the E2 ubiquitin conjugating (UBC) enzyme. The viral ubiquitin-conjugating enzyme (UBCv1) is expressed throughout ASFV infection and accumulates at late times post infection. UBCv is also present in the viral particle suggesting that the ubiquitin-proteasome pathway could play an important role at early ASFV infection. We determined that inhibition of the final stage of the ubiquitin-proteasome pathway blocked a post-internalization step in ASFV replication in Vero cells. Under proteasome inhibition, ASF viral genome replication, late gene expression and viral production were severely reduced. Also, ASFV enhanced proteasome activity at late times and the accumulation of polyubiquitinated proteins surrounding viral factories. Core-associated and/or viral proteins involved in DNA replication may be targets for the ubiquitin-proteasome pathway that could possibly assist virus uncoating at final core breakdown and viral DNA release. At later steps, polyubiquitinated proteins at viral factories could exert regulatory roles in cell signaling.
[Mh] Termos MeSH primário: Vírus da Febre Suína Africana/genética
Febre Suína Africana/genética
Enzimas de Conjugação de Ubiquitina/genética
Proteínas Virais/genética
Replicação Viral/genética
[Mh] Termos MeSH secundário: Febre Suína Africana/virologia
Vírus da Febre Suína Africana/patogenicidade
Animais
Cercopithecus aethiops
Replicação do DNA/genética
DNA Viral/genética
Genoma Viral
Complexo de Endopeptidases do Proteassoma/genética
Suínos/virologia
Ubiquitina/genética
Células Vero
Vírion/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA, Viral); 0 (Ubiquitin); 0 (Viral Proteins); EC 2.3.2.23 (Ubiquitin-Conjugating Enzymes); EC 2.3.2.23 (ubiquitin-conjugating enzyme UBCv1, African swine fever virus); EC 3.4.25.1 (Proteasome Endopeptidase Complex)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180102
[Lr] Data última revisão:
180102
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171216
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0189741


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[PMID]:29235978
[Au] Autor:Hakobyan A; Galindo I; Nañez A; Arabyan E; Karalyan Z; Chistov AA; Streshnev PP; Korshun VA; Alonso C; Zakaryan H
[Ad] Endereço:1​Group of Antiviral Defense Mechanisms, Institute of Molecular Biology of NAS RA, 0014, Yerevan, Armenia.
[Ti] Título:Rigid amphipathic fusion inhibitors demonstrate antiviral activity against African swine fever virus.
[So] Source:J Gen Virol;99(1):148-156, 2018 Jan.
[Is] ISSN:1465-2099
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Rigid amphipathic fusion inhibitors (RAFIs) are a family of nucleoside derivatives that inhibit the infectivity of several enveloped viruses by interacting with virion envelope lipids and inhibiting fusion between viral and cellular membranes. Here we tested the antiviral activity of two RAFIs, 5-(Perylen-3-ylethynyl)-arabino-uridine (aUY11) and 5-(Perylen-3-ylethynyl)uracil-1-acetic acid (cm1UY11) against African swine fever virus (ASFV), for which no effective vaccine is available. Both compounds displayed a potent, dose-dependent inhibitory effect on ASFV infection in Vero cells. The major antiviral effect was observed when aUY11 and cm1UY11 were added at early stages of infection and maintained during the complete viral cycle. Furthermore, virucidal assay revealed a significant extracellular anti-ASFV activity for both compounds. We also found decrease in the synthesis of early and late viral proteins in Vero cells treated with cm1UY11. Finally, the inhibitory effect of aUY11 and cm1UY11 on ASFV infection in porcine alveolar macrophages was confirmed. Overall, our study has identified novel anti-ASFV compounds with potential for future therapeutic developments.
[Mh] Termos MeSH primário: Vírus da Febre Suína Africana/efeitos dos fármacos
Antivirais/farmacologia
Perileno/análogos & derivados
Uracila/análogos & derivados
Uridina/análogos & derivados
Proteínas Virais/antagonistas & inibidores
Vírion/efeitos dos fármacos
Internalização do Vírus/efeitos dos fármacos
[Mh] Termos MeSH secundário: Vírus da Febre Suína Africana/crescimento & desenvolvimento
Vírus da Febre Suína Africana/metabolismo
Animais
Antivirais/síntese química
Membrana Celular/efeitos dos fármacos
Membrana Celular/virologia
Cercopithecus aethiops
Relação Dose-Resposta a Droga
Macrófagos Alveolares/efeitos dos fármacos
Macrófagos Alveolares/virologia
Testes de Sensibilidade Microbiana
Perileno/síntese química
Perileno/farmacologia
Cultura Primária de Células
Suínos
Uracila/síntese química
Uracila/farmacologia
Uridina/síntese química
Uridina/farmacologia
Células Vero
Proteínas Virais/biossíntese
Vírion/crescimento & desenvolvimento
Vírion/metabolismo
Replicação Viral/efeitos dos fármacos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (5-(perylen-3-yl)ethynylarabinouridine); 0 (5-(perylen-3-ylethynyl)uracil-1-acetic acid); 0 (Antiviral Agents); 0 (Viral Proteins); 56HH86ZVCT (Uracil); 5QD5427UN7 (Perylene); WHI7HQ7H85 (Uridine)
[Em] Mês de entrada:1712
[Cu] Atualização por classe:171229
[Lr] Data última revisão:
171229
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171214
[St] Status:MEDLINE
[do] DOI:10.1099/jgv.0.000991


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[PMID]:28768556
[Au] Autor:Nielsen JP; Larsen TS; Halasa T; Christiansen LE
[Ad] Endereço:Dynamical Systems, Department of Applied Mathematics and Computer Sciences,Technical University of Denmark,Lyngby,Denmark.
[Ti] Título:Estimation of the transmission dynamics of African swine fever virus within a swine house.
[So] Source:Epidemiol Infect;145(13):2787-2796, 2017 10.
[Is] ISSN:1469-4409
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The spread of African swine fever virus (ASFV) threatens to reach further parts of Europe. In countries with a large swine production, an outbreak of ASF may result in devastating economic consequences for the swine industry. Simulation models can assist decision makers setting up contingency plans. This creates a need for estimation of parameters. This study presents a new analysis of a previously published study. A full likelihood framework is presented including the impact of model assumptions on the estimated transmission parameters. As animals were only tested every other day, an interpretation was introduced to cover the weighted infectiousness on unobserved days for the individual animals (WIU). Based on our model and the set of assumptions, the within- and between-pen transmission parameters were estimated to ß w = 1·05 (95% CI 0·62-1·72), ß b = 0·46 (95% CI 0·17-1·00), respectively, and the WIU = 1·00 (95% CI 0-1). Furthermore, we simulated the spread of ASFV within a pig house using a modified SEIR-model to establish the time from infection of one animal until ASFV is detected in the herd. Based on a chosen detection limit of 2·55% equivalent to 10 dead pigs out of 360, the disease would be detected 13-19 days after introduction.
[Mh] Termos MeSH primário: Vírus da Febre Suína Africana/fisiologia
Febre Suína Africana/transmissão
[Mh] Termos MeSH secundário: Febre Suína Africana/diagnóstico
Febre Suína Africana/virologia
Animais
Abrigo para Animais
Modelos Teóricos
Suínos
Reino Unido
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1709
[Cu] Atualização por classe:171125
[Lr] Data última revisão:
171125
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170804
[St] Status:MEDLINE
[do] DOI:10.1017/S0950268817001613


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[PMID]:28721858
[Au] Autor:Abworo EO; Onzere C; Oluoch Amimo J; Riitho V; Mwangi W; Davies J; Blome S; Peter Bishop R
[Ad] Endereço:1​International Livestock Research Institute (ILRI), P.O. Box 30709, GPO 00100, Nairobi, Kenya.
[Ti] Título:Detection of African swine fever virus in the tissues of asymptomatic pigs in smallholder farming systems along the Kenya-Uganda border: implications for transmission in endemic areas and ASF surveillance in East Africa.
[So] Source:J Gen Virol;98(7):1806-1814, 2017 Jul.
[Is] ISSN:1465-2099
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The persistence of African swine fever virus (ASFV) in endemic areas, with small-scale but regular outbreaks in domestic pigs, is not well understood. ASFV has not been detected using conventional diagnosis in these pigs or adjacent populations of resistant African wild pigs, that could act as potential carriers during the outbreaks. However, such data are crucial for the design of evidence-based control strategies. We conducted cross-sectional (1107 pigs) and longitudinal (100 pigs) monitoring of ASFV prevalence in local pigs in Kenya and Uganda. The horizontal survey revealed no evidence of ASFV in the serum or blood using either conventional or real-time PCR. One pig consistently tested positive using ELISA, but negative using PCR assays on blood. Interestingly, the isotype of the antibodies from this animal were strongly IgA biased relative to control domestic pigs and warthogs, suggesting a role for mucosal immunity. The tissues from this pig were positive by PCR following post-mortem. Internal organ tissues of 44 healthy pigs (28 sentinel pigs and 16 pigs from slaughter slabs) were tested with four different PCR assays; 15.9 % were positive for ASFV suggesting that healthy pigs carrying ASFV exist in the swine population in the study area. P72 and p54 genotyping of ASFV revealed very limited diversity: all were classified in genotype IX at both loci, as were virtually all viruses causing recent ASF outbreaks in the region. Our study suggests that carrier pigs may play a role in ASF disease outbreaks, although the triggers for outbreaks remain unclear and require further investigation. This study significantly increases scientific knowledge of the epidemiology of ASF in the field in Africa, which will contribute to the design of effective surveillance and control strategies.
[Mh] Termos MeSH primário: Vírus da Febre Suína Africana/isolamento & purificação
Febre Suína Africana/virologia
[Mh] Termos MeSH secundário: África Oriental/epidemiologia
Febre Suína Africana/diagnóstico
Febre Suína Africana/epidemiologia
Febre Suína Africana/transmissão
Vírus da Febre Suína Africana/classificação
Vírus da Febre Suína Africana/genética
Criação de Animais Domésticos
Animais
Doenças Assintomáticas
Estudos Transversais
Surtos de Doenças
Genótipo
Quênia/epidemiologia
Suínos
Uganda/epidemiologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170921
[Lr] Data última revisão:
170921
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170720
[St] Status:MEDLINE
[do] DOI:10.1099/jgv.0.000848


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[PMID]:28502836
[Au] Autor:Portugal RS; Bauer A; Keil GM
[Ad] Endereço:Institut für molekulare Virologie und Zellbiologie, Friedrich-Loeffler-Institut, Südufer 10, Greifswald, Insel Riems 17493, Germany. Electronic address: portmari@gmail.com.
[Ti] Título:Selection of differently temporally regulated African swine fever virus promoters with variable expression activities and their application for transient and recombinant virus mediated gene expression.
[So] Source:Virology;508:70-80, 2017 Aug.
[Is] ISSN:1096-0341
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:African swine fever virus threatens pig production worldwide due to the lack of vaccines, for which generation of both deletion and insertion mutants is considered. For development of the latter, operational ASFV promoters of different temporal regulation and strengths are desirable. We therefore compared the capacities of putative promoter sequences from p72, CD2v, p30, viral DNA polymerase and U104L genes to mediate expression of luciferase from transfected plasmids after activation in trans, or p30-, DNA polymerase- and U104L promoters in cis, using respective ASFV recombinants. We identified sequences with promoter activities upstream the viral ORFs, and showed that they differ in both their expression intensity regulating properties and in their temporal regulation. In summary, p30 and DNA polymerase promoters are recommended for high level early regulated transgene expression. For late expression, the p72, CD2v and U104L promoter are suitable. The latter however, only if low level transgene expression is aimed.
[Mh] Termos MeSH primário: Vírus da Febre Suína Africana/genética
Febre Suína Africana/virologia
Regiões Promotoras Genéticas
[Mh] Termos MeSH secundário: Vírus da Febre Suína Africana/metabolismo
Animais
Expressão Gênica
Genes Reporter
Luciferases/genética
Luciferases/metabolismo
Suínos
Transfecção
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
EC 1.13.12.- (Luciferases)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170818
[Lr] Data última revisão:
170818
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170516
[St] Status:MEDLINE


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[PMID]:28494931
[Au] Autor:Karalyan ZR; Ter-Pogossyan ZR; Karalyan NY; Semerjyan ZB; Tatoyan MR; Karapetyan SA; Karalova EM
[Ad] Endereço:Laboratory of Cell Biology and Virology, Institute of Molecular Biology of NAS RA, 0014 Yerevan, Armenia; Yerevan State Medical University, Armenia. Electronic address: zkaralyan@yahoo.com.
[Ti] Título:Hemophagocytic lymphohistiocytosis in acute African swine fever clinic.
[So] Source:Vet Immunol Immunopathol;187:64-68, 2017 May.
[Is] ISSN:1873-2534
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:INTRODUCTION: Hemophagocytic lymphohistiocytosis (HLH) usually has been defined as the combination of a proliferation of cytologically benign, actively phagocytic macrophages in bone marrow, spleen, lymph nodes, etc. in association with fever, cytopenia, splenomegaly, and hypertriglyceridemia. HLH is often triggered by viral infection. The aim of this study was to ascertain the features of HLH involvement in African swine fever virus (ASFV) (genotype II) pathogenesis. METHODS: The serum levels of macrophage colony-stimulating factor (MCSF) and granulocyte-macrophage colony-stimulating factor (GMCSF), as well as the histological constitution (for hemophagocytic macrophages detection) of various organs of pigs infected with ASFV genotype II were investigated. The diagnosis of HLH was made according to universally accepted human criteria. RESULTS: The association of fever, cytopenias, splenomegaly, and hemophagocytosis was present in 87.5% of the infected pigs (absence of hyperthermia in one of eight pigs). Marked hypertriglyceridemia was observed at 3-4days post infection. Previously it was shown that ASFV induced a significant decrease in the level of fibrinogen from day 5 till the end of experiment. Progression of the HLH coincided with a temporary increase in the serum levels of MCSF levels (early stage of disease) and GMCSF levels (2-3 pays post infection). CONCLUSIONS: Hemophagocytic syndrome should be suspected in ASFV (genotypeII) infected pigs.
[Mh] Termos MeSH primário: Febre Suína Africana/etiologia
Linfo-Histiocitose Hemofagocítica/veterinária
[Mh] Termos MeSH secundário: Febre Suína Africana/imunologia
Febre Suína Africana/patologia
Vírus da Febre Suína Africana/imunologia
Animais
Ensaio de Imunoadsorção Enzimática/veterinária
Pulmão/patologia
Linfo-Histiocitose Hemofagocítica/complicações
Linfo-Histiocitose Hemofagocítica/imunologia
Linfo-Histiocitose Hemofagocítica/patologia
Suínos
Triglicerídeos/sangue
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Triglycerides)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170929
[Lr] Data última revisão:
170929
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170513
[St] Status:MEDLINE


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[PMID]:28481911
[Au] Autor:Lokhandwala S; Waghela SD; Bray J; Sangewar N; Charendoff C; Martin CL; Hassan WS; Koynarski T; Gabbert L; Burrage TG; Brake D; Neilan J; Mwangi W
[Ad] Endereço:Department of Veterinary Pathobiology, Texas A&M University, College Station, TX, United States of America.
[Ti] Título:Adenovirus-vectored novel African Swine Fever Virus antigens elicit robust immune responses in swine.
[So] Source:PLoS One;12(5):e0177007, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:African Swine Fever Virus (ASFV) is a high-consequence transboundary animal pathogen that often causes hemorrhagic disease in swine with a case fatality rate close to 100%. Lack of treatment or vaccine for the disease makes it imperative that safe and efficacious vaccines are developed to safeguard the swine industry. In this study, we evaluated the immunogenicity of seven adenovirus-vectored novel ASFV antigens, namely A151R, B119L, B602L, EP402RΔPRR, B438L, K205R and A104R. Immunization of commercial swine with a cocktail of the recombinant adenoviruses formulated in adjuvant primed strong ASFV antigen-specific IgG responses that underwent rapid recall upon boost. Notably, most vaccinees mounted robust IgG responses against all the antigens in the cocktail. Most importantly and relevant to vaccine development, the induced antibodies recognized viral proteins from Georgia 2007/1 ASFV-infected cells by IFA and by western blot analysis. The recombinant adenovirus cocktail also induced ASFV-specific IFN-γ-secreting cells that were recalled upon boosting. Evaluation of local and systemic effects of the recombinant adenovirus cocktail post-priming and post-boosting in the immunized animals showed that the immunogen was well tolerated and no serious negative effects were observed. Taken together, these outcomes showed that the adenovirus-vectored novel ASFV antigen cocktail was capable of safely inducing strong antibody and IFN-γ+ cell responses in commercial swine. The data will be used for selection of antigens for inclusion in a multi-antigen prototype vaccine to be evaluated for protective efficacy.
[Mh] Termos MeSH primário: Adenoviridae/genética
Vírus da Febre Suína Africana/genética
Febre Suína Africana/imunologia
Antígenos Virais/imunologia
Suínos/imunologia
[Mh] Termos MeSH secundário: Vírus da Febre Suína Africana/imunologia
Animais
Antígenos Virais/genética
Western Blotting
Ensaio de Imunoadsorção Enzimática
Vetores Genéticos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antigens, Viral)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170915
[Lr] Data última revisão:
170915
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170509
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0177007


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[PMID]:28418311
[Au] Autor:van Heerden J; Malan K; Gadaga BM; Spargo RM
[Ti] Título:Reemergence of African Swine Fever in Zimbabwe, 2015.
[So] Source:Emerg Infect Dis;23(5):860-861, 2017 05.
[Is] ISSN:1080-6059
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Zimbabwe is the only country in southern Africa with no reported African swine fever (ASF) outbreaks during 1993-2014. However, the 2015 discovery of genotype II ASF virus in Zimbabwe indicates the reemergence of ASF in this country and suggests that this viral genotype may be spreading through eastern and southern Africa.
[Mh] Termos MeSH primário: Vírus da Febre Suína Africana/genética
Febre Suína Africana/epidemiologia
Febre Suína Africana/virologia
Doenças Transmissíveis Emergentes
[Mh] Termos MeSH secundário: Febre Suína Africana/história
Vírus da Febre Suína Africana/classificação
Animais
Surtos de Doenças
Genes Virais
História do Século XXI
Filogenia
Análise de Sequência de DNA
Suínos
Zimbábue/epidemiologia
[Pt] Tipo de publicação:HISTORICAL ARTICLE; JOURNAL ARTICLE
[Em] Mês de entrada:1706
[Cu] Atualização por classe:171109
[Lr] Data última revisão:
171109
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170419
[St] Status:MEDLINE
[do] DOI:10.3201/eid2305.161195


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[PMID]:28381576
[Au] Autor:Frouco G; Freitas FB; Coelho J; Leitão A; Martins C; Ferreira F
[Ad] Endereço:Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Lisbon, Portugal.
[Ti] Título:DNA-Binding Properties of African Swine Fever Virus pA104R, a Histone-Like Protein Involved in Viral Replication and Transcription.
[So] Source:J Virol;91(12), 2017 Jun 15.
[Is] ISSN:1098-5514
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:African swine fever virus (ASFV) codes for a putative histone-like protein (pA104R) with extensive sequence homology to bacterial proteins that are implicated in genome replication and packaging. Functional characterization of purified recombinant pA104R revealed that it binds to single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) over a wide range of temperatures, pH values, and salt concentrations and in an ATP-independent manner, with an estimated binding site size of about 14 to 16 nucleotides. Using site-directed mutagenesis, the arginine located in pA104R's DNA-binding domain, at position 69, was found to be relevant for efficient DNA-binding activity. Together, pA104R and ASFV topoisomerase II (pP1192R) display DNA-supercoiling activity, although none of the proteins by themselves do, indicating that the two cooperate in this process. In ASFV-infected cells, A104R transcripts were detected from 2 h postinfection (hpi) onward, reaching a maximum concentration around 16 hpi. pA104R was detected from 12 hpi onward, localizing with viral DNA replication sites and being found exclusively in the Triton-insoluble fraction. Small interfering RNA (siRNA) knockdown experiments revealed that pA104R plays a critical role in viral DNA replication and gene expression, with transfected cells showing lower viral progeny numbers (up to a reduction of 82.0%), lower copy numbers of viral genomes (-78.3%), and reduced transcription of a late viral gene (-47.6%). Taken together, our results strongly suggest that pA104R participates in the modulation of viral DNA topology, probably being involved in viral DNA replication, transcription, and packaging, emphasizing that ASFV mutants lacking the A104R gene could be used as a strategy to develop a vaccine against ASFV. Recently reintroduced in Europe, African swine fever virus (ASFV) causes a fatal disease in domestic pigs, causing high economic losses in affected countries, as no vaccine or treatment is currently available. Remarkably, ASFV is the only known mammalian virus that putatively codes for a histone-like protein (pA104R) that shares extensive sequence homology with bacterial histone-like proteins. In this study, we characterized the DNA-binding properties of pA104R, analyzed the functional importance of two conserved residues, and showed that pA104R and ASFV topoisomerase II cooperate and display DNA-supercoiling activity. Moreover, pA104R is expressed during the late phase of infection and accumulates in viral DNA replication sites, and its downregulation revealed that pA104R is required for viral DNA replication and transcription. These results suggest that pA104R participates in the modulation of viral DNA topology and genome packaging, indicating that A104R deletion mutants may be a good strategy for vaccine development against ASFV.
[Mh] Termos MeSH primário: Vírus da Febre Suína Africana/genética
Vírus da Febre Suína Africana/fisiologia
Proteínas de Ligação a DNA/química
Proteínas de Ligação a DNA/metabolismo
Histonas/química
Histonas/metabolismo
Transcrição Genética
Replicação Viral
[Mh] Termos MeSH secundário: Febre Suína Africana/prevenção & controle
Animais
Cercopithecus aethiops
Proteínas de Ligação a DNA/genética
Proteínas de Ligação a DNA/isolamento & purificação
Expressão Gênica
Genoma Viral
Histonas/genética
Mutagênese Sítio-Dirigida
Reação em Cadeia da Polimerase em Tempo Real
Deleção de Sequência
Sus scrofa/virologia
Suínos
Células Vero
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA-Binding Proteins); 0 (Histones)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:171126
[Lr] Data última revisão:
171126
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170407
[St] Status:MEDLINE


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[PMID]:28298603
[Au] Autor:Eaton HE; Kobayashi T; Dermody TS; Johnston RN; Jais PH; Shmulevitz M
[Ad] Endereço:Department of Medical Microbiology and Immunology, Li Ka Shing Institute of Virology, University of Alberta, Edmonton, Alberta, Canada.
[Ti] Título:African Swine Fever Virus NP868R Capping Enzyme Promotes Reovirus Rescue during Reverse Genetics by Promoting Reovirus Protein Expression, Virion Assembly, and RNA Incorporation into Infectious Virions.
[So] Source:J Virol;91(11), 2017 Jun 01.
[Is] ISSN:1098-5514
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Reoviruses, like many eukaryotic viruses, contain an inverted 7-methylguanosine (m7G) cap linked to the 5' nucleotide of mRNA. The traditional functions of capping are to promote mRNA stability, protein translation, and concealment from cellular proteins that recognize foreign RNA. To address the role of mRNA capping during reovirus replication, we assessed the benefits of adding the African swine fever virus NP868R capping enzyme during reovirus rescue. C3P3, a fusion protein containing T7 RNA polymerase and NP868R, was found to increase protein expression 5- to 10-fold compared to T7 RNA polymerase alone while enhancing reovirus rescue from the current reverse genetics system by 100-fold. Surprisingly, RNA stability was not increased by C3P3, suggesting a direct effect on protein translation. A time course analysis revealed that C3P3 increased protein synthesis within the first 2 days of a reverse genetics transfection. This analysis also revealed that C3P3 enhanced processing of outer capsid µ1 protein to µ1C, a previously described hallmark of reovirus assembly. Finally, to determine the rate of infectious-RNA incorporation into new virions, we developed a new recombinant reovirus S1 gene that expressed the fluorescent protein UnaG. Following transfection of cells with UnaG and infection with wild-type virus, passage of UnaG through progeny was significantly enhanced by C3P3. These data suggest that capping provides nontraditional functions to reovirus, such as promoting assembly and infectious-RNA incorporation. Our findings expand our understanding of how viruses utilize capping, suggesting that capping provides nontraditional functions to reovirus, such as promoting assembly and infectious-RNA incorporation, in addition to enhancing protein translation. Beyond providing mechanistic insight into reovirus replication, our findings also show that reovirus reverse genetics rescue is enhanced 100-fold by the NP868R capping enzyme. Since reovirus shows promise as a cancer therapy, efficient reovirus reverse genetics rescue will accelerate production of recombinant reoviruses as candidates to enhance therapeutic potency. NP868R-assisted reovirus rescue will also expedite production of recombinant reovirus for mechanistic insights into reovirus protein function and structure.
[Mh] Termos MeSH primário: Vírus da Febre Suína Africana/enzimologia
Nucleotidiltransferases/metabolismo
Orthoreovirus de Mamíferos/genética
Orthoreovirus de Mamíferos/fisiologia
RNA Viral/metabolismo
Vírion/fisiologia
Montagem de Vírus
[Mh] Termos MeSH secundário: Vírus da Febre Suína Africana/genética
Linhagem Celular
Proteínas Recombinantes de Fusão/metabolismo
Genética Reversa
Vírion/genética
Replicação Viral
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Viral); 0 (Recombinant Fusion Proteins); EC 2.7.7.- (Nucleotidyltransferases); EC 2.7.7.50 (mRNA guanylyltransferase)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:171112
[Lr] Data última revisão:
171112
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170317
[St] Status:MEDLINE



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