Suppression of p53 point mutations by mdm-2
One microgram of pCOSXl-CAT is cotransfected with 50 ng of p53 expression plasmids encoding either human wild-type (Wt) or p53 double-point mutants into SAOS-2 cells. Five micrograms of mdm-2 expression plasmid encoding the suppressive mutant Δ222–437 or vector alone are also cotransfected. The CAT activities induced by each p53 plasmid in the presence of vector are set at 100%. Each mutant was analyzed at least five times. Error bars represent standard deviations. Experiments using full-length mdm-2 produced similar results (data not shown). The p53 double-point mutant 2/3 contains 2E-K, 3E-K; mutant 14/19 contains 14L-Q, 19F-S; mutant 25/26 contains 25L-Q, 26L-H substitutions.