Related Experiment Videos
Host-dependent temperature-sensitive growth of HVJ (Sendai virus) wild-type in rat glioma C 6 cells
Abstract:
At non-permissive temperature viral specific RNA synthesis was not restricted in rat glioma (C 6) cells infected with HVJ (Sendai virus) wild-type. However, as has previously been shown (J Gen Virol [1984] 65: 639-643), the synthesis of M protein was reduced at non-permissive temperature, in contrast to the L, P, HN, Fo and NP proteins which were synthesized in comparable amounts at permissive and non-permissive temperatures. In this report we show additionally that viral nucleocapsids (NC), which consist of L, P and NP proteins, were formed within the infected cells at both temperatures. Hemagglutinin and neuraminidase activities were also detected in samples incubated at non-permissive temperature. By membrane immunofluorescence and cell-surface immunoprecipitation it was shown that migration of HN and Fo proteins to the cell surface occurred normally at non-permissive temperature. Additionally, the L, P and NP proteins, which were associated with the plasma membrane isolated from the infected cells maintained at permissive temperature, were absent from the membrane of cells incubated at non-permissive temperature. These results suggest that NC and glycoproteins synthesized at non-permissive temperature could not assemble effectively at the plasma membrane because of a lack of M protein. Thus, the host-dependent ts lesion of HVJ in C 6 cells was considered to be mainly in M protein synthesis.
Insights
The host-dependent temperature-sensitive lesion in HVJ (Sendai virus) in C6 cells is primarily due to M protein synthesis defects, hindering viral assembly at non-permissive temperatures.
Area of Science:
- Virology
- Molecular Biology
- Cell Biology
Background:
- HVJ (Sendai virus) exhibits temperature-sensitive (ts) behavior in C6 rat glioma cells.
- Previous studies indicated reduced M protein synthesis at non-permissive temperatures.
Purpose of the Study:
- To investigate the host-dependent ts lesion of HVJ in C6 cells.
- To elucidate the role of M protein in viral assembly and cell surface expression.
Main Methods:
- Infection of C6 cells with wild-type HVJ at permissive and non-permissive temperatures.
- Analysis of viral RNA and protein synthesis (L, P, HN, Fo, NP, M).
- Detection of viral nucleocapsids (NC), hemagglutinin, and neuraminidase activities.
- Membrane immunofluorescence and cell-surface immunoprecipitation.
Main Results:
- Viral RNA synthesis was not restricted at non-permissive temperatures.
- M protein synthesis was significantly reduced, while other viral proteins were synthesized comparably.
- Viral nucleocapsids formed at both temperatures.
- Glycoproteins (HN, Fo) reached the cell surface, but L, P, and NP proteins were absent from the plasma membrane at non-permissive temperatures.
- Defective M protein synthesis impaired the assembly of viral components at the plasma membrane.
Conclusions:
- The ts lesion in HVJ-infected C6 cells is primarily located in M protein synthesis.
- Lack of M protein prevents effective assembly of viral nucleocapsids and glycoproteins at the plasma membrane.
- M protein is crucial for the proper localization and assembly of viral components in host cells.