Key result
Regeneration of infectious Harvey viruses from truncated proviruses involves illegitimate recombination of cellular or cotransfected DNAs flanking the 5'-LTR-ras gene, rather than RNA template switching.
Retroviral transduction of oncogenic sequences proceeds via rare illegitimate recombinations between proviral and cellular DNAs rather than RNA template switching.
Challenges prior recombination models in retrovirology; leaves open relevance to human gene therapy vectors.
We have studied whether the origin of retroviral onc genes, by transduction of sequences from cellular proto-onc genes, involves DNA or RNA recombination. By using altered Harvey sarcoma proviruses as models for transduction intermediates, we have investigated the mechanism of regeneration of transforming virus from truncated proviruses with only a single 5' long terminal repeat (LTR) but with a complete 5'-LTR-ras transforming gene. The Harvey proviruses were specifically altered to discriminate between virus regeneration by RNA template switching during reverse transcription, as has been postulated, and virus regeneration by DNA recombination with either helper virus or among elements of the defective provirus alone. For this purpose U3 elements of the Harvey proviral LTR, which are essential for replication but not for transcription, were deleted in vitro. Only proviral constructions with an intact or a nearly intact single LTR regenerated infectious Harvey sarcoma virus. Since all constructions transformed cells and produced identical RNAs, our results exclude a model of virus regeneration by switching of RNA templates during reverse transcription. We conclude that regeneration of infectious Harvey viruses from truncated provirus involved illegitimate recombination of cellular or cotransfected DNAs flanking the 5'-LTR-ras gene of Harvey sarcoma virus. Based on this and evidence from the literature, we propose that retroviral transduction proceeds by way of rare illegitimate recombinations between proviral and cellular DNAs.
No takes yet. Share an insight, caveat, or question.
Goodrich et al. (1988) studied this question. Altered Harvey sarcoma proviruses vs. Wild-type Harvey provirus or other constructs was evaluated on Regeneration of infectious Harvey sarcoma virus. Regeneration of infectious Harvey viruses from truncated proviruses involves illegitimate recombination of cellular or cotransfected DNAs flanking the 5'-LTR-ras gene, rather than RNA template switching.
Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context: