Enhanced induction of SV40 replication from transformed rat cells by fusion with UV-irradiated normal and repair-deficient human fibroblasts.
Fusion of SV40-transformed rat (BRKSV) cells which do not spontaneously produce infectious virus, with permissive monkey cells resulted in a low level of production of infectious virus in the heterokaryons. UV-Irradiation of the BRKSV cells prior to fusion did not result in increased virus production, but irradiation of the monkey cells prior to fusion did result in enhanced induction (EI) of SV40, as compared to control experiments in which neither cell type was irradiated. This indicated that rat cells lack the ability to initiate replication of integrated SV40 upon UV-irradiation and do not contain "permissiveness" factors that are required to support SV40 replication. In contrast, monkey cells do contain such permissiveness factors which seem to be temporally enhanced by UV-irradiation, and thus may be responsible for the EI phenomenon. Expression of EI was dose-dependent and reached maximum values approximately 24 h after UV-irradiation. The kinetics of EI resembled that of EI previously established for SV40 induction in semi-permissive cells, and of enhanced reactivation (ER) and enhanced mutagenesis (EM) of SV40 in monkey cells. Similar kinetics of EI were obtained when human diploid fibroblasts were used for fusion with BRKSV cells. Similar levels of EI were found with normal human cells and repair-deficient xeroderma pigmentosum (XP) cells of complementation groups A and C, and XP variant cells. This suggests that expression of EI is not related to excision repair. Since EI is also normally expressed in XP cells which display an abnormal ER of HSV and in XP variant cells which show a delayed EM of HSV, we conclude that EI may occur independently of ER and EM. Finally it was shown that treatment of human cells with N-ethyl-N-nitrosourea results in similar induction of EI as irradiation with UV-light, and that addition of TPA in fusion experiments has no effect on EI.
D000818:Animals; D002459:Cell Fusion; D002472:Cell Transformation, Viral; D002478:Cells, Cultured / Q000528:radiation effects; D002522:Chlorocebus aethiops; D004260:DNA Repair; D005038:Ethylnitrosourea / Q000494:pharmacology; D006801:Humans; D051381:Rats; D013539:Simian virus 40 / Q000235:genetics*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors; D014779:Virus Replication / Q000187:drug effects / Q000528:radiation effects
D013755:Tetradecanoylphorbol Acetate; D005038:Ethylnitrosourea