TsJT60, a cell cycle G0- ts mutant, becomes lethal at non-permissive temperature by transformation with adenovirus 5 when the expression of E1B gene is lacking

Abstract

tsJT60, a temperature-sensitive ( ts) mutant cell line of Fischer rat, is viable at both permissive (34 °C) and non-permissive (39.5 °C) temperatures. The cells grow normally in exponential growth phase at both temperatures, but when stimulated with fetal bovine serum (FBS) from G0 phase they re-enter S phase at 34 °C but not at 39.5 °. When tsJT60 cells were transformed with adenovirus (Ad) 5 wild type, they grew well at both temperatures, expressed E1A and E1B genes, and formed colonies in soft agar. When tsJT60 cells were transformed with Ad 5 dl313, that lacks E1B gene, the transformed cells grew well at 34 °C but failed to form colony in soft agar. They died very soon at 39.5 °C. 3Y1 cells (a parental line of tsJT60) transformed with dl313 grew well at both temperatures, although neither expressed E1B gene nor formed colonies in soft agar. The phenotype of being lethal at 39.5 °C of dl313-transformed tsJT60 cells was complemented by cell fusion with 3Y1BU r cells (5-BrdU-resistant 3Y1), but not with tsJT60TG r cells (6-thioguanine resistant tsJT60). These results indicate that the lethal phenotype is related to the ts mutation of tsJT60 cells and also to the deletion of E1B gene of Ad5.