The Experts below are selected from a list of 446178 Experts worldwide ranked by ideXlab platform
Eladio Vinuela - One of the best experts on this subject based on the ideXlab platform.
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characterization of the african swine fever virus Structural Protein p14 5 a dna binding Protein
Virology, 1997Co-Authors: Luisa Martinezpomares, Carmen Simonmateo, Carlos Lopezotin, Eladio VinuelaAbstract:Abstract The gene encoding the Structural Protein p14.5 of African swine fever virus (ASFV) has been mapped and sequenced. This gene, designated E120R, is located in theSalI H/EcoRI E restriction fragment of the ASFV genome and is predicted to encode a Protein of 120 amino acids with a molecular weight of 13.4 kDa. Northern-blot analysis showed that E120R is transcribed at late times during the viral replication cycle. The E120R gene product has been expressed inEscherichia coli,purified, and used as an antigen for antibody production. The antiserum anti-pE120R recognized a Protein in infected cell extracts with an apparent molecular mass of 14.5 kDa, named p14.5. This antiserum also detected Protein p14.5 in purified virus particles. Protein p14.5 is synthesized late in infection and is located in viral factories. Immunoprecipitation analysis and binding-assay experiments have shown that Protein p14.5 interacts with a Protein that could correspond to the major Structural Protein p72. Purified Protein p14.5 interacts with DNA in a sequence-independent manner. It binds to both single-stranded and double-stranded DNA. A possible role of Protein p14.5 in the encapsidation of ASFV DNA is suggested.
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high level expression in escherichia coli of the gene coding for the major Structural Protein p72 of african swine fever virus
Gene, 1993Co-Authors: Jose M P Freije, Eladio Vinuela, M Munoz, Carlos LopezotinAbstract:Abstract The gene encoding the major Structural Protein (p72) of African swine fever virus (ASFV) has been expressed in Escherichia coli using a T7 RNA polymerase system. The use of a recombinant plasmid which contains the entire gene inserted between the T7 promoter and the transcription terminator of the expression vector allowed us to obtain a high expression level of the intact viral Protein. This polypeptide, which appears in the insoluble fraction of the bacterial extracts, showed an intense reaction with the antibodies present in the sera of ASFV-infected animals, as demonstrated by Western blot and enzyme-linked immunosorbent assay. The recombinant Protein was purified by size-exclusion highperformance liquid chromatography and used to develop a serological test of the disease.
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Protein p22 of african swine fever virus an early Structural Protein that is incorporated into the membrane of infected cells
Virology, 1991Co-Authors: Ana Camacho, Eladio VinuelaAbstract:Abstract The open reading frame K'177, located at the left end of the African swine fever virus genome, codes for an early induced Structural Protein of 22,000 Da (p22), which is released from the viral particle by the nonionic detergent n -octyl-β- d -glucopyranoside under conditions that solubilize external viral Structural Proteins. The predicted amino acid sequence of the Protein contains a hydrophobic region at its N-terminus with characteristics of a signal peptide and, at early times after virus infection of Vero cells, the Protein can be detected in the cell membrane by immunolabeling.
Naoto Ito - One of the best experts on this subject based on the ideXlab platform.
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severe acute respiratory syndrome coronavirus 3a Protein is a viral Structural Protein
Journal of Virology, 2005Co-Authors: Naoto Ito, Eric C Mossel, Krishna Narayanan, Clarence J. Peters, Taisuke Inoue, Vsevolod L. Popov, Cheng Huang, Shinji MakinoAbstract:The present study showed the association of a severe acute respiratory syndrome coronavirus (SCoV) accessory Protein, 3a, with plasma membrane and intracellular SCoV particles in infected cells. 3a Protein appeared to undergo posttranslational modifications in infected cells and was incorporated into SCoV particles, establishing that 3a Protein was a SCoV Structural Protein.
Piet A Van Rijn - One of the best experts on this subject based on the ideXlab platform.
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Structural Protein vp2 of african horse sickness virus is not essential for virus replication in vitro
Journal of Virology, 2017Co-Authors: Rene G P Van Gennip, Piet A Van Rijn, Sandra G P Van De Water, Christiaan A PotgieterAbstract:The Reoviridae family consists of nonenveloped multilayered viruses with a double-stranded RNA genome consisting of 9 to 12 genome segments. The Orbivirus genus of the Reoviridae family contains African horse sickness virus (AHSV), bluetongue virus, and epizootic hemorrhagic disease virus, which cause notifiable diseases and are spread by biting Culicoides species. Here, we used reverse genetics for AHSV to study the role of outer capsid Protein VP2, encoded by genome segment 2 (Seg-2). Expansion of a previously found deletion in Seg-2 indicates that Structural Protein VP2 of AHSV is not essential for virus replication in vitro. In addition, inframe replacement of RNA sequences in Seg-2 by that of green fluorescence Protein (GFP) resulted in AHSV expressing GFP, which further confirmed that VP2 is not essential for virus replication. In contrast to virus replication without VP2 expression in mammalian cells, virus replication in insect cells was strongly reduced, and virus release from insect cells was completely abolished. Further, the other outer capsid Protein, VP5, was not copurified with virions for virus mutants without VP2 expression. AHSV without VP5 expression, however, could not be recovered, indicating that outer capsid Protein VP5 is essential for virus replication in vitro. Our results demonstrate for the first time that a Structural viral Protein is not essential for orbivirus replication in vitro, which opens new possibilities for research on other members of the Reoviridae family.
Luis Enjuanes - One of the best experts on this subject based on the ideXlab platform.
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The envelope Protein of severe acute respiratory syndrome coronavirus interacts with the non-Structural Protein 3 and is ubiquitinated.
Virology, 2010Co-Authors: Enrique Alvarez, Jose L. Nieto-torres, Jose M. Jimenez-guardeño, Laura Marcos-villar, Marta L Dediego, Luis EnjuanesAbstract:Abstract To analyze the Proteins interacting with the severe acute respiratory syndrome coronavirus (SARS-CoV) envelope (E) Protein, a SARS-CoV was engineered including two tags associated to the E Protein. Using this virus, complexes of SARS-CoV E and other Proteins were purified using a tandem affinity purification system. Several viral and cell Proteins including spike, membrane, non-Structural Protein 3 (nsp3), dynein heavy chain, fatty acid synthase and transmembrane Protein 43 bound E Protein. In the present work, we focused on the binding of E Protein to nsp3 in infected cells and cell-free systems. This interaction was mediated by the N-terminal acidic domain of nsp3. Moreover, nsp3 and E Protein colocalized during the infection. It was shown that E Protein was ubiquitinated in vitro and in cell culture, suggesting that the interaction between nsp3 and E Protein may play a role in the E Protein ubiquitination status and therefore on its turnover.
Espen Rimstad - One of the best experts on this subject based on the ideXlab platform.
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The non-Structural Protein μNS of piscine orthoreovirus (PRV) forms viral factory-like structures
Veterinary Research, 2016Co-Authors: Hanne Merethe Haatveit, Maria Krudtaa Dahle, Turhan Markussen, Ingvild B. Nyman, Øystein Wessel, Espen RimstadAbstract:AbstractPiscine orthoreovirus (PRV) is associated with heart- and skeletal muscle inflammation in farmed Atlantic salmon. The virus is ubiquitous and found in both farmed and wild salmonid fish. It belongs to the family Reoviridae, closely related to the genus Orthoreovirus. The PRV genome comprises ten double-stranded RNA segments encoding at least eight Structural and two non-Structural Proteins. Erythrocytes are the major target cells for PRV. Infected erythrocytes contain globular inclusions resembling viral factories; the putative site of viral replication. For the mammalian reovirus (MRV), the non-Structural Protein μNS is the primary organizer in factory formation. The analogous PRV Protein was the focus of the present study. The subcellular location of PRV μNS and its co-localization with the PRV σNS, µ2 and λ1 Proteins was investigated. We demonstrated that PRV μNS forms dense globular cytoplasmic inclusions in transfected fish cells, resembling the viral factories of MRV. In co-transfection experiments with μNS, the σNS, μ2 and λ1 Proteins were recruited to the globular structures. The ability of μNS to recruit other PRV Proteins into globular inclusions indicates that it is the main viral Protein involved in viral factory formation and pivotal in early steps of viral assembly.
