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Transformation of human cells by a polyomavirus containing complementing JCV and RFV genomes
F J O'Neill1, L Renzetti, T H Miller
1Research Service, VA Medical Center, Salt Lake City, Utah.
Virus Research
|September 1, 1988
Summary
A novel infectious virus, E-JCV/L-RFV, was created by combining early JCV and late RFV DNA. This virus partially transformed human cells, indicating a role for L-RFV in initiating transformation.
Area of Science:
- Virology
- Molecular Biology
- Oncology
Background:
- Human polyomaviruses, including John Cunningham virus (JCV), are associated with various diseases.
- The role of different viral components in cellular transformation is not fully understood.
Purpose of the Study:
- To investigate the transforming potential of early JCV (E-JCV) and late RFV (L-RFV) DNA.
- To determine the role of L-RFV in the transformation process initiated by E-JCV.
Main Methods:
- Molecular cloning of viral DNA from E-JCV and L-RFV.
- Transfection of cloned DNA into human fetal brain (HFB) and human embryonic kidney (HEK) cells.
- Complementation assays to produce infectious viral particles.
- Analysis of cellular transformation, T-antigen expression, and viral DNA presence in transformants.
Main Results:
- A new infectious virus, E-JCV/L-RFV, was produced through complementation.
- Infection with E-JCV/L-RFV resulted in partial transformation of HFB and HEK cells.
- Transformants expressed T-antigen and had an extended lifespan, but showed incomplete transformation phenotypes.
- E-JCV DNA was detected in transformants, while L-RFV DNA was absent, yet L-RFV was implicated in transformation initiation.
Conclusions:
- The combination of E-JCV and L-RFV can generate a transforming virus.
- L-RFV plays a critical role in the initiation of cellular transformation, even in the absence of its own DNA in the final transformants.
- This study sheds light on the complex interplay of viral components in oncogenesis.