RESUMEN
The virus capsomeres of the outer and inner layers of capsids were effectively released simultaneously from purified virions by lipase digestion and were purified by a linear gradient ultracentrifugation. The capsid consisted of an array of double layers of uniformly arranged individual capsomeres where a lipid(s) served as a matrix in between the capsomeres.
Asunto(s)
Virus de la Fiebre Porcina Africana/análisis , Cápside/aislamiento & purificación , Iridoviridae/análisis , Virus de la Fiebre Porcina Africana/ultraestructura , Cápside/ultraestructura , Lipasa , Microscopía Electrónica , Virión/análisis , Virión/ultraestructuraRESUMEN
Frog erythrocytic virus (FEV), one of the largest icosahedral viruses, is enveloped, measures up to 450 nm in diameter, and contains double stranded DNA. The virus is found in the cytoplasm of erythrocytes of Rana catesbeiana, Rana septentrionalis, and Rana clamitans from Algonquin Park, Ontario (Canada). Acidophilic inclusions in infected erythrocytes stained with Giemsa's stain correspond to viroplasms from which FEV buds and forms aggregates of virus particles as seen in the electron microscope. Frog erythrocytic virus appears to acquire its envelope from lamellar membranes which surround the virus particles. The virus is structurally sensitive to cesium chloride, potassium tartrate and glycerol. It is also sensitive at pH 1 to 5 and a temperature of 56 C for 15 min. The virus contains at least 16 proteins which range in relative molecular mass from 19.5 to 91.0 kilodaltons (kDa), with two major proteins of 31.0 and 43.0 kDa. The viral DNA has a buoyant density of 1.690 +/- 0.005 g/ml, guanine plus cytosine ratio of 25 to 36%, and a melting temperature of 82 to 86 C. Data from this study indicate that FEV should be included in the family Iridoviridae.
Asunto(s)
Eritrocitos/microbiología , Iridoviridae/ultraestructura , Ranidae , Virosis/veterinaria , Animales , Citoplasma/microbiología , ADN Viral/análisis , Eritrocitos/ultraestructura , Cuerpos de Inclusión Viral/ultraestructura , Iridoviridae/análisis , Iridoviridae/genética , Microscopía Electrónica , Ontario , Prevalencia , Rana catesbeiana , Proteínas Virales/análisis , Virosis/epidemiología , Virosis/microbiologíaRESUMEN
Highly purified African swine fever virus contains a cyclic AMP-independent protein kinase which phosphorylates endogenous virus proteins with a specific activity of about 0.45 pmol/microgram of virus protein. The major substrates for the virion protein kinase in vitro were the structural proteins p10 and p9. Both proteins were phosphorylated preferentially at serine residues. A possible relationship between protein p10 phosphorylation and RNA synthesis in vitro by the virion-associated RNA polymerase is suggested by the finding that N-alpha-tosyl-L-lysyl-chloromethyl ketone inhibited both phosphorylation of p10 and transcription. Two phosphoproteins, with molecular masses of 35 and 17 kDa, were found in African swine fever virus purified from infected Vero cells labeled with [32P]phosphate. A phosphopolypeptide with a molecular mass of about 35 kDa was found in the cytoplasm of infected Vero cells.
Asunto(s)
Virus de la Fiebre Porcina Africana/análisis , Iridoviridae/análisis , Fosfoproteínas/metabolismo , Proteínas Quinasas/metabolismo , Proteínas Virales/metabolismo , Adenosina Trifosfato/metabolismo , Virus de la Fiebre Porcina Africana/efectos de los fármacos , Virus de la Fiebre Porcina Africana/enzimología , Animales , Electroforesis en Gel de Poliacrilamida , Peso Molecular , Octoxinol , Fosforilación , Fosfotreonina/análisis , Fosfotreonina/metabolismo , Polietilenglicoles/farmacología , ARN Viral/biosíntesis , ARN Viral/efectos de los fármacos , Clorometilcetona Tosilisina/farmacología , Células Vero , Virión/análisisRESUMEN
At least 74 acidic and 37 basic proteins are synthesized in African swine fever virus (ASFV)-infected monkey cells not detected in uninfected cells analyzed by two-dimensional gel electrophoresis. Essentially all the proteins synthesized early during infection are also observed at late times. The use of inhibitors such as cycloheximide and phosphonoacetate has led to the identification of 34 immediate early and 13 delayed early polypeptides. Therefore 64 proteins were classified as late polypeptides. Several ASFV-induced proteins are phosphorylated as proteins a1, a4, a20, a41, a48, a49, a51, a52, a55, a58, a67, b2, b12, b28, and b32.
Asunto(s)
Virus de la Fiebre Porcina Africana/análisis , Iridoviridae/análisis , Proteínas Virales/biosíntesis , Virus de la Fiebre Porcina Africana/efectos de los fármacos , Cicloheximida/farmacología , Electroforesis en Gel de Poliacrilamida , Ácido Fosfonoacético/farmacología , Fosfoproteínas/biosíntesis , Procesamiento Proteico-Postraduccional , Factores de TiempoRESUMEN
Seven African swine fever virus structural proteins were localized in the virion by immunoelectron microscopy. African swine fever virus-infected cells were incubated, before or after embedding and thin sectioning, with monoclonal antibodies specific for different structural proteins, and after labeling with protein A-gold complexes, the samples were examined in the electron microscope. Proteins p14 and p24 were found in the external region of the virion, proteins p12, p72, p17, and p37 were found in the intermediate layers, and protein p150 was found in the nucleoid and in one vertex. A monoclonal antibody that recognized protein p150 as well as p220, a virus-induced, nonstructural protein, could also bind to a component present in the nucleus of both uninfected and virus-infected cells.
Asunto(s)
Virus de la Fiebre Porcina Africana/análisis , Iridoviridae/análisis , Proteínas Virales/análisis , Virus de la Fiebre Porcina Africana/ultraestructura , Animales , Anticuerpos Monoclonales , Línea Celular , Núcleo Celular/análisis , Chlorocebus aethiops , Técnicas Inmunológicas , Microscopía Electrónica , Modelos Biológicos , Proteínas Virales/inmunología , Proteínas Estructurales Virales , Virión/análisisRESUMEN
Image analysis of freeze-etch replicas of cylindrical aberrant forms of FV3 provided evidence for three morphological subunits protruding from the six-coordinated capsomers. Negatively stained capsomers displayed both triangular and hexagonal profiles which suggests that their innermost portion is pseudohexagonal. Images from underfocused micrographs of capsomers are indicative of a central channel. The trimeric nature of the capsomer has been established by electrophoresis in the presence of Triton X-100, which showed that the molecular weight of the nondissociated capsomer is about 140,000 whereas that of the polypeptide itself is 48,000. This trimeric association does not occur via disulfide bonds, and inside the capsomers there are no free amino groups accessible to the usual bifunctional reagents. Thus, the chemical nature of the interpolypeptide bonds inside the trimers is still unknown. We have previously estimated the triangulation number (T) of FV3 to be 147 or 133 (Darcy-Tripier et al., 1984). The present study, using optical diffraction of the facets of FV3, allowed a better determination of the angle of skewness and is in favor of T = 133 (h = 9, k = 4, 18 degrees).
Asunto(s)
Cápside , Iridoviridae/ultraestructura , Fenómenos Químicos , Química , Densitometría , Electroforesis en Gel de Poliacrilamida , Grabado por Congelación , Iridoviridae/análisis , Sustancias Macromoleculares , Microscopía Electrónica , Peso Molecular , Coloración y EtiquetadoRESUMEN
Analysis of highly purified fish Lymphocystis Disease Virus (LDV), strain Leetown NFH, by three different methods, namely periodic Acid Schiff reaction, radiolabelling with tritiated fucose and N-acetyl-D-glucosamine and staining with three lectins, indicated that ten glycoproteins were associated with the virus structure. Six of them were detected by all of the three methods, three by both radiolabelling and lectin staining but only one by the lectin technique. Localization of these glycoproteins at the surface or inside the virion is discussed.
Asunto(s)
Peces/microbiología , Iridoviridae/análisis , Proteínas Virales/análisis , Animales , Glicoproteínas/análisis , Lectinas , Peso Molecular , Reacción del Ácido Peryódico de Schiff , Proteínas del Envoltorio Viral/análisisAsunto(s)
Iridoviridae/fisiología , Animales , Células Cultivadas , ADN Viral/análisis , Enfermedades de los Peces/microbiología , Peces/microbiología , Iridoviridae/análisis , Iridoviridae/enzimología , Iridoviridae/ultraestructura , Proteínas Virales/análisis , Virosis/microbiología , Virosis/veterinaria , Replicación ViralAsunto(s)
Virus de la Fiebre Porcina Africana , Iridoviridae , Fiebre Porcina Africana/inmunología , Fiebre Porcina Africana/microbiología , Fiebre Porcina Africana/fisiopatología , Virus de la Fiebre Porcina Africana/análisis , Virus de la Fiebre Porcina Africana/clasificación , Virus de la Fiebre Porcina Africana/inmunología , Virus de la Fiebre Porcina Africana/fisiología , Animales , Anticuerpos Antivirales , Antígenos Virales/inmunología , ADN Viral/análisis , ADN Viral/biosíntesis , Iridoviridae/análisis , Iridoviridae/clasificación , Iridoviridae/inmunología , Iridoviridae/fisiología , Macrófagos/microbiología , Monocitos/microbiología , Porcinos , Proteínas Virales/análisis , Proteínas Virales/biosíntesis , Proteínas Estructurales Virales , Replicación ViralAsunto(s)
Iridoviridae , Lepidópteros/microbiología , Animales , Antígenos Virales/análisis , Línea Celular , Efecto Citopatogénico Viral , ADN Viral/análisis , Hidrolasas/metabolismo , Insectos/microbiología , Iridoviridae/análisis , Iridoviridae/enzimología , Iridoviridae/fisiología , Iridoviridae/ultraestructura , Lípidos/análisis , Proteínas Virales/análisis , Replicación ViralAsunto(s)
Crustáceos/microbiología , Iridoviridae/fisiología , Nematodos/microbiología , Animales , Antígenos Virales/inmunología , Reacciones Cruzadas , Crustáceos/parasitología , ADN Viral/análisis , Iridoviridae/análisis , Iridoviridae/inmunología , Iridoviridae/ultraestructura , Microscopía Electrónica , Morfogénesis , Rickettsia/crecimiento & desarrollo , Proteínas Virales/análisisRESUMEN
Thirty-seven monoclonal antibodies to seven frog virus 3 (FV3) structural proteins were isolated and used to examine the distribution of viral proteins within virions and infected cells. Three monoclonal antibodies, one to the major capsid protein, VP55, and two to VP38 had detectable neutralizing activity suggesting that these proteins are located on the surface of virions. Immunofluorescent studies showed that VP108, VP57, VP55, and VP16 were localized mainly within virus assembly sites, while VP17 was detected in both assembly sites and the surrounding cytoplasm. The abundance of viral structural proteins within assembly sites is consistent with the idea that virion maturation occurs exclusively within assembly sites.
Asunto(s)
Iridoviridae/análisis , Proteínas Virales/análisis , Virión/análisis , Animales , Anticuerpos Monoclonales , Línea Celular , Cricetinae , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente , Riñón , Ratones , Ratones Endogámicos BALB C , Peso MolecularRESUMEN
Molecules of DNA of Aedes cantans mosquito iridescent virus were found to be of linear shape, about 150 micron in length. The temperature of melting, sedimentation coefficient, molecular weight, and buoyant density of DNA were determined as well as the content of GC pairs in it.
Asunto(s)
Aedes/microbiología , ADN Viral/aislamiento & purificación , Iridoviridae/aislamiento & purificación , Animales , Fenómenos Químicos , Química Física , ADN Viral/análisis , Iridoviridae/análisis , Cinética , Microscopía ElectrónicaAsunto(s)
Virus de la Fiebre Porcina Africana , Iridoviridae , Fiebre Porcina Africana/diagnóstico , Virus de la Fiebre Porcina Africana/análisis , Animales , Enzimas de Restricción del ADN/metabolismo , ADN Viral/análisis , Desoxirribonucleasa EcoRI , Ensayo de Inmunoadsorción Enzimática , Iridoviridae/análisis , Microscopía Electrónica , Porcinos , Proteínas Virales/análisisRESUMEN
African Swine Fever virus infected MS cells labeled with radioactive 14C-amino acids, 32Pi or [3H]-glucosamine were examined by high resolution sodium dodecylsulfate polyacrylamide gel electrophoresis and showed 43 infected cell polypeptides. Twenty-one of these proteins were present in the nuclear fraction of infected cells. At least 22 of the infected cell polypeptides induced antibodies during natural infections in swine. The pattern of infected cell polypeptides modified by incorporation of showed prosthetic groups that at least 8 polypeptides were phosphorylated and at least three specific viral glycoproteins (A, B and C) were detected by immunoprecipitation. The most highly glycosylated polypeptide corresponds to the structural viral protein VP51.
Asunto(s)
Virus de la Fiebre Porcina Africana/análisis , Glicoproteínas/análisis , Iridoviridae/análisis , Fosfoproteínas/análisis , Proteínas Virales/análisis , Virus de la Fiebre Porcina Africana/fisiología , Animales , Anticuerpos Antivirales/biosíntesis , Antígenos Virales/inmunología , Línea Celular , Núcleo Celular/análisis , Haplorrinos , Riñón , Proteínas Virales/inmunología , Proteínas Estructurales ViralesRESUMEN
The Chilo Iridescent Virus (CIV) is a lipid-containing virus propagated in vitro in choristoneura fumiferana cell cultures. We have analysed the individual lipids of the viral membrane which appeared interesting in their relative amounts and mainly in the high proportion of phosphatidylinositol. This fraction represented about 27 per cent of the phospholipid extract. The lipid composition of the viral membrane was unchanged whether the virus was propagated in vivo in larvae or in vitro in invertebrate cell cultures and was clearly different from that of the hosts.
Asunto(s)
Iridoviridae/análisis , Lípidos/análisis , Ácidos Grasos/análisis , Fosfatidilinositoles/análisis , Fosfolípidos/análisisRESUMEN
Frog virus 3 inoculated into mice induces an acute degenerative hepatitis. This hepatitis is of toxic origin since the virus is unable to multiply at 37 degrees C. The Kupffer cells, which are the target cells for FV3, reveal the presence of viral particles, viral DNA and proteins. Although the hepatocytes present early and drastic nuclear lesions, viral particles were never observed in these cells. Viral proteins however but not DNA, could be found inside parenchymal cells.