The fragmentation of incoming Semliki Forest virus nucleocapsids in mosquito (Aedes albopictus) cells might be coupled to virion uncoating.
The fate of Semliki Forest virus (SFV) nucleocapsid, especially the capsid protein (C-protein), was investigated during the early stages of a productive infection in mosquito Aedes albopictus cells. Infection of the cells resulted in a time dependent accumulation of a C-protein derived fragment. This fragmentation of incoming viral nucleocapsid was prevented by NH4Cl, an agent generally used to elevate the pH in acidic intracellular compartments, suggesting that a low intravesicular pH is required for this process. Density gradient analysis of the postnuclear cell lysate demonstrated that the fragmentation was associated with a cellular compartment showing a density of 1.14 +/- 0.02 g/ml. This cellular compartment was devoid from a lysosomal marker enzyme and represented the timely preceding cellular fraction through which SFV passed before encountering a lysosomal fraction. Furthermore, the intracellular distribution of the viral, 3H-uridine-labeled RNA suggested that the same fraction might represent a key cellular compartment in which the separation of the viral RNA from the viral structural proteins is primed. In conclusion, these data lead to the suggestion that the fragmentation of incoming SFV nucleocapsids in Aedes albopictus cells might be the part of the mechanism leading to the release of viral RNA into the cytosol during early stages of productive infection.
Mrkic
B
B
Institute of Biochemistry, University of Bern, Switzerland.
Kempf
C
C
D000330:Aedes / Q000166:cytology; D000818:Animals; D002460:Cell Line; D002500:Centrifugation, Isopycnic; D007553:Isotope Labeling; D019251:Nucleocapsid / Q000502:physiology*; D012367:RNA, Viral / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism; D012672:Semliki forest virus / Q000235:genetics / Q000502:physiology*; D014316:Tritium; D014529:Uridine; D014771:Virion; D019065:Virus Assembly
D012367:RNA, Viral; D014316:Tritium; D012260:Ribonucleases; D014529:Uridine