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Characterization of ribonucleoproteins and ribosomes isolated from lymphocytic choriomeningitis virus
Abstract:
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.
Insights
Researchers isolated two lymphocytic choriomeningitis virus (LCMV) nucleoproteins (RNPs) and ribosomal subunits. These viral RNPs, associated with viral RNA, showed partial sensitivity to RNase, offering insights into LCMV structure.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Lymphocytic choriomeningitis virus (LCMV) is a significant human pathogen.
- Understanding the molecular composition of LCMV is crucial for developing antiviral strategies.
Purpose of the Study:
- To isolate and characterize the nucleoproteins (RNPs) of LCMV.
- To investigate the association of viral RNA with these RNPs.
- To compare viral RNPs with host ribosomal subunits.
Main Methods:
- Disruption of purified LCMV using Nonidet P-40 and KCl.
- Sucrose gradient centrifugation for isolation of viral components.
- Buoyant density determination in CsCl.
- RNase sensitivity assays.
Main Results:
- Two distinct viral RNPs were isolated, sedimenting at 123S-148S and 83S.
- The larger RNP was associated with 23S and 31S viral RNA; the smaller with 23S viral RNA.
- Viral RNPs had a buoyant density of 1.32 g/cm³, distinct from isolated 40S and 60S ribosomal subunits.
- Viral RNPs exhibited partial sensitivity to RNase.
Conclusions:
- LCMV possesses distinct nucleoprotein complexes associated with its RNA genome.
- These RNPs share some properties with host ribosomal subunits but are structurally unique.
- The partial RNase sensitivity suggests a specific RNA-protein interaction within the viral RNP structure.