Products of reverse transcription in avian retrovirus analyzed by electron microscopy

DNA products synthesized in avian retroviral particles permeabilized with melittin have been analyzed in an electron microscope. These studies have provided further insight and subsequent refinement in the melittin activation techniques. Our electron microscope analyses verify the existence of the plus-strand single-stranded DNA branches, presumed to originate by strand-displacement synthesis (L. R. Boone and A. M. Skalka, J. Virol. 37:117-126, 1981). The branches occur at many locations along the DNA molecules and are observed at very early times, even before the minus-strand copies of the RNA genome are completed. Circular forms of different derivations are observed at early and at late times, which are possible intermediates in viral replication. Novel forms termed H structures are also described. In addition to the identification of possible intermediates, these analyses have provided further information on the sequence of events in retroviral reverse transcription. These new data are combined with previous results to generate a model of reverse transcription which incorporates strand-displacement synthesis as an essential feature.