Effect of gamma radiation on retroviral recombination.
To elucidate the mechanism(s) of retroviral recombination, we exposed virions to gamma radiation prior to infecting target cells. By using previously described spleen necrosis virus-based vectors containing multiple markers, recombinant proviruses were studied after a single round of retrovirus replication. The current models of retroviral recombination predict that breaking virion RNA should promote minus-strand recombination (forced copy-choice model), decrease or not affect plus-strand recombination (strand displacement/assimilation model), and shift plus-strand recombination towards the 3' end of the genome. However, we found that while gamma irradiation of virions reduced the amount of recoverable viral RNA, it did not primarily cause breaks. Thus, the frequency of selected recombinants was not significantly altered with greater doses of radiation. In spite of this, the irradiation did decrease the number of recombinants with only one internal template switch. As a result, the average number of additional internal template switches in the recombinant proviruses increased from 0.7 to 1.4 as infectivity decreased to 6%. The unselected internal template switches tended to be 5' of the selected crossover even in the recombinants from irradiated viruses, inconsistent with a plus-strand recombination mechanism.
Hu
W S
WS
McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.
Temin
H M
HM
D001483:Base Sequence; D015152:Blotting, Northern; D002478:Cells, Cultured; D005720:Gamma Rays; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification / Q000528:radiation effects; D011995:Recombination, Genetic / Q000528:radiation effects; D012190:Retroviridae / Q000235:genetics* / Q000528:radiation effects
D012367:RNA, Viral