The introduction of dominant-negative p53 mutants suppresses temperature shift-induced senescence in immortal human fibroblasts expressing a thermolabile SV40 large T antigen

Immortal human fibroblasts, SVts8 cells, which express a heat-labile SV40 l arge T antigen, induces a senescence-like phenomenon in response to upward shift in temperature. Cells with arrested division show strong induction of senescence-associated beta-galactosidase, We examined how p53 and pRB are involved in this phenomenon since they are major targets of the T antigen. Transfection of cells with plasmids encoding the wild-type T antigen or hum an papilloma virus type 16 E6/E7 proteins completely abolished the arrest i n cell division, a plasmid encoding the E6 protein suppressed it markedly, while a plasmid encoding E7 had no effect. Plasmids encoding dominant-negat ive p53 mutants also suppressed the arrest in cell division to various degr ees. Upon temperature shift, p21 mRNA was upregulated 10-fold in SVts8 cell s, but only slightly in clones expressing the wild-type T antigen or domina nt-negative p53 mutants. These data demonstrate that p53 plays a major role in this senescence-like phenomenon.