We studied Moloney murine leukemia virus replication in newly infected Balb c/3T3 cells brought to the G0 phase by serum depletion. Using the polymerase chain reaction method, we showed that Moloney murine leukemia virus can be efficiently internalized in nonproliferating fibroblasts, although reverse transcription of the viral RNA in these cells remains incomplete. It seems likely that a lower availability of deoxyribonucleotides in G0-arrested cells is responsible for this premature termination of the reverse transcription step. Accordingly, the addition of high concentrations of nucleosides to the culture medium of nondividing cells simultaneously with infection enables them to complete the reverse transcription process, without re-initiating the cell cycle. Inhibition of reverse transcription by hydroxyurea confirms the dependence of this retroviral step on the intracellular nucleotide pool rather than on the precise arrest point of the host cell cycle. Furthermore, the pyrimidine nucleotide pool, and more particularly the cytidine pool, appears to play a central regulatory role in this step.
Animals Base Sequence