Cellular functions are required for the synthesis and integration of avian sarcoma virus-specific DNA

Abstract

We have used two experimental strategies to test the role of cellular functions in the synthesis and integration of virus-specific DNA in cells infected by avian sarcoma virus. First, quail embryo fibroblasts, placed in stationary phase (G 0) by prolonged serum starvation, did not support the efficient synthesis of viral DNA during the first 24–48 hr after infection. Synthesis of viral DNA was impaired according to at least two parameters: the amount of DNA was diminished, particularly the amount of the plus-strand DNA (identical in polarity to the viral genome); and the length of both minus and plus strands was reduced in the stationary cells. In parallel cultures fed with fresh serum, over two thirds of the cells were able to reenter the cell cycle within 24 hr, and viral DNA of normal size was synthesized. Second, density labeling of viral and cellular DNA with BUdR was used to determine whether cellular DNA synthesis was required for integration of viral DNA. In both quail embryo fibroblasts released from G 0 by serum replacement and randomly growing duck embryo fibroblasts, viral DNA was integrated only into cellular DNA replicated during the infection. Our results indicate that serum-starved cells lack a factor (or factors) required for the efficient and complete synthesis of ASV-specific DNA. We have not been able to establish whether such factor(s) are present in growing cells only during S phase. Integration of viral DNA appears to require cellular DNA synthesis; this may be due to a requirement for a factor (or factors) present in adequate concentration only during S phase or to a requirement for the structural changes in cellular DNA that accompany replication.