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Studies with aphidicolin on the Fv-1 host restriction of Friend murine leukemia virus
Abstract:
The murine gene Fv-1 exerts a major control over the replication of Friend murine leukemia virus (F-MuLV). An effect of the gene product has been determined to be at the level of accumulation and integration of viral DNA. Aphidicolin, an inhibitor of eucaryotic DNA polymerase alpha, was studied in murine cells infected either permissively or nonpermissively with regard to the Fv-1 genotype. Results indicated that inhibition of DNA polymerase alpha did not affect the accumulation of form III viral DNA in either permissive or nonpermissive cells. However, the normal accumulation of circular form I DNA in permissive cells was inhibited. The block in the accumulation of form I DNA resembled that occurring in some F-MuLV Fv-1-nonpermissive infections. Additionally, aphidicolin treatment resulted in the accumulation of novel low-molecular-weight viral DNA species, normally detectable in a nonpermissive infection of NIH cells with B-tropic F-MuLV. These data suggest that the Fv-1 gene product may interact with host DNA polymerase alpha to prevent viral replication.
Insights
The Fv-1 gene controls Friend murine leukemia virus (F-MuLV) replication by interacting with host DNA polymerase alpha. Inhibiting this enzyme affects viral DNA accumulation, suggesting a key role in viral restriction.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- The Fv-1 gene in mice significantly influences Friend murine leukemia virus (F-MuLV) replication.
- Viral DNA accumulation and integration are key steps affected by the Fv-1 gene product.
Purpose of the Study:
- To investigate the role of host DNA polymerase alpha in Fv-1-mediated restriction of F-MuLV.
- To determine if inhibiting DNA polymerase alpha mimics or modifies the Fv-1 restriction phenotype.
Main Methods:
- Murine cells with different Fv-1 genotypes (permissive and nonpermissive) were infected with F-MuLV.
- Cells were treated with aphidicolin, a specific inhibitor of eukaryotic DNA polymerase alpha.
- Viral DNA forms (form III, form I, and low-molecular-weight species) were analyzed.
Main Results:
- Aphidicolin did not affect the accumulation of form III viral DNA in either permissive or nonpermissive cells.
- Inhibition of DNA polymerase alpha blocked the accumulation of circular form I viral DNA in permissive cells.
- Aphidicolin treatment led to the accumulation of novel low-molecular-weight viral DNA species, similar to those seen in nonpermissive infections.
Conclusions:
- The Fv-1 gene product likely interacts with host DNA polymerase alpha.
- This interaction appears crucial for preventing viral replication, particularly at the stage of viral DNA form I accumulation.
- DNA polymerase alpha is implicated as a potential target or interacting partner in Fv-1 restriction.