Structural components and replication of arenaviruses.
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Biomedical subjects
Publications and source records attributed to I R Pedersen.
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Disruption of purified lymphocytic choriomeningitis (LCM) virus with Nonidet P-40 in 0.5 M KCl followed by sucrose gradient centrifugation in 0.3 M KCl led to the isolation of two viral nucleoproteins (RNPs) as well as 40S and 60S ribosomal subunits. The largest viral RNP sedimented heterogenously at 123S to 148S and was associated with 23S and 31S viral RNA. The other viral RNP sedimented at 83S and was associated with 23S viral RNA. The buoyant density in CsCl was determined to be 1.32 g/cm3 for the viral RNP. Densities of 1.52 and 1.60 g/cm3 were determined for the 40S and 60S subunits, similar to those of the BHK-21 cells subunits dissociated by 0.5 M KCl. The viral RNPs were partly sensitive to RNase.
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Purified preparations of lymphocytic choriomeningitis virus (LCM virus) contain three classes of RNA. The previously described 18s, 23s, 28s, and 31s RNAs, where the 23s and 31s RNAs are viral-specific, and the 18s and 28s RNAs probably are host RNAs incorporated in the virion. Now, 4s, 5s, and 5.5s RNAs can be isolated as well. Thus five RNAs which migrate by acrylamide gel electrophoresis as ribosome-derived RNA can be isolated from purified LCM virus. This observation further supports the reports that arenaviruses may contain ribosomes. The ribosome-derived RNA can be synthesized both before and after the virus infection. The viral 23s could be a hydrogen-bonded complex forming the 31s RNA, or it could be contained in defective interfering LCM virus particles; these possibilities are examined.
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Lymphocytic choriomeningitis (LCM) virus, Traub strain, was purified from BHK-21 tissue culture medium. The virus was then analyzed by equilibrium centrifugation and rate zonal centrifugation in sucrose gradients. A buoyant density in sucrose of 1.18 g/ml was found and the S(20, w) value was estimated to be about 470 to 500S. Furthermore, the (3)H-uridine-labeled ribonucleic acid (RNA) from virus was extracted from LCM virus and analyzed by rate zonal centrifugation. Two major and one minor single-stranded RNA components were found with sedimentation coefficients of 28, 22, and 18S.
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