RESUMEN
Cell-associated proteins of rodent (CG-1820 strain) and human (K-27 and 360 strains) hantaviruses isolated in the European endemic areas of the U.S.S.R. are antigenically similar as revealed by immunoprecipitation and immunoblotting assays. Nucleocapsid-associated RNAs of representative hantaviruses of four antigenic groups [Sugiyama et al. (1987) J Gen Virol 68: 979-987] have unique PAGE patterns. "Electropherotyping" of the RNAs isolated from infected cells might be used for identifying and distinguishing hantaviruses.
Asunto(s)
Fiebre Hemorrágica con Síndrome Renal/microbiología , Orthohantavirus/análisis , ARN Viral/análisis , Proteínas Virales/análisis , Animales , Cápside/análisis , Fiebre Hemorrágica con Síndrome Renal/inmunología , Humanos , Técnicas de Inmunoadsorción , Peso Molecular , Pruebas de Precipitina , U.R.S.S. , Células Vero , Proteínas del Núcleo Viral/análisisRESUMEN
Single-stranded RNAs from Lassa virus-infected cells were fractionated by affinity chromatography on an oligo(dT)-cellulose column as well as by sucrose gradient centrifugation. Fractionated RNAs were translated in rabbit reticulocyte lysates, and translation products were precipitated with monoclonal antibodies to the nucleocapsid protein of Lassa virus. It has been shown that mRNA for the nucleocapsid protein is 15-16S and the RNA does not contain long if any poly(A) sequences.
Asunto(s)
Arenaviridae/genética , Cápside/genética , Virus Lassa/genética , ARN Mensajero/aislamiento & purificación , ARN Viral/aislamiento & purificación , Proteínas del Núcleo Viral/genética , Animales , Fiebre de Lassa/microbiología , Poli A/genética , Poli A/aislamiento & purificación , Biosíntesis de Proteínas , ARN Mensajero/genética , ARN Viral/genética , Células VeroRESUMEN
RNA from Machupo virus infected cells was centrifuged in a linear sucrose gradient and RNAs from gradient fractions were tested separately for template activity in a cell-free protein synthesizing system from rabbit reticulocytes. Fraction 15-16 S programmed the synthesis of protein that migrated in SDS-polyacrylamide gel like the nucleocapsid protein of a purified virus. The synthesis of virus glycoproteins was not detected in the system.
Asunto(s)
Arenaviridae/genética , Arenavirus del Nuevo Mundo/genética , Biosíntesis de Proteínas , ARN Mensajero/genética , ARN Viral/genética , Animales , Arenavirus del Nuevo Mundo/metabolismo , Cápside/biosíntesis , Cápside/genética , Células Vero , Proteínas del Núcleo Viral/biosíntesis , Proteínas del Núcleo Viral/genética , Proteínas Virales/biosíntesis , Proteínas Virales/genéticaRESUMEN
Sucrose gradient velocity centrifugation, polyacrylamide gel electrophoresis and RNA-RNA hybridization were used to characterize Lassa and Machupo virion RNAs as well as virus-specific RNAs from cells infected with Pichinde and Machupo viruses. Five RNA species: 30-31S, 28S, 22-24S, 18S and 4-6S have been detected in Lassa, Machupo, and Pichinde virion RNAs. Among them 28S, 18S and 4-6S RNAs cosediment and comigrate with respectively cell RNAs. RNase resistance analyses suggest the presence of extensive secondary structures and complementary RNAs in Lassa, Machupo, and Pichinde virion RNAs. Annealing with poly(A)-containing RNA from infected cells has revealed that the bulk of "minus" strands of Machupo virion RNA is located in 22-24S and 28-31S fractions of sucrose gradient. Thus Machupo and Lassa viruses as well as Pichinde virus contain two genomic RNA fragments: "large" (molecular weight of about 2.2 X 10(6] and "small" (molecular weight of about 1.3 X 10(6]. In the cells infected with Pichinde virus and treated with actinomycin D (1.0 microgram/ml) synthesis of 18S, 22-24S and 30-31S RNAs has been registered. At least 22-24S and 30-31S classes comprise "plus" and "minus" strands. In cells infected with Machupo virus in the presence of actinomycin D the synthesis of similar sedimentation classes of RNAs and certain amounts of 28S RNA have been detected.