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Visualization of cytoskeletal elements and associated retroviral antigens by immunogold transmission electron
Abstract:
Several investigators have reported an association between the cytoskeleton and viral antigens. In our laboratory, biochemical immunofluorescence and immuno-gold electron microscopy studies were conducted on TX-100 extracted NIH/3T3 cells infected with Moloney-murine leukemia virus. Cytochalasin B treatment causes reversible microfilament disruption and a concomitant decrease in virus production. No effect on microtubules was seen. Immunogold electron microscopy reveals an association between cytoskeletal actin and the viral antigens gp70 and p15E. The results of these immunocytological and biochemical studies indicate that the cytoskeleton may play an integral role in transport and processing of viral gene-envelope products.
Insights
The cytoskeleton, specifically actin filaments, is linked to Moloney-murine leukemia virus production. Disrupting microfilaments with Cytochalasin B reduces viral output, suggesting a role in viral transport.
Area of Science:
- Virology
- Cell Biology
- Immunology
Background:
- An association between the cytoskeleton and viral antigens has been previously reported.
- The specific role of cytoskeletal components in viral replication remains an area of investigation.
Purpose of the Study:
- To investigate the role of the cytoskeleton in the replication of Moloney-murine leukemia virus.
- To determine if cytoskeletal integrity affects viral antigen processing and production.
Main Methods:
- Biochemical analysis
- Immunofluorescence microscopy
- Immuno-gold electron microscopy
- TX-100 extraction of NIH/3T3 cells infected with Moloney-murine leukemia virus
- Cytochalasin B treatment to disrupt microfilaments
Main Results:
- Cytochalasin B treatment caused reversible microfilament disruption.
- A decrease in virus production was observed following microfilament disruption.
- No effect was observed on microtubules.
- Immunogold electron microscopy showed an association between cytoskeletal actin and viral antigens gp70 and p15E.
Conclusions:
- The cytoskeleton, particularly actin filaments, plays an integral role in the transport and processing of viral gene-envelope products.
- Cytoskeletal integrity is important for efficient Moloney-murine leukemia virus production.