Mutant p53 proteins behave in a dominant, negative fashion in vivo.
The p53 encodes a cellular phosphoprotein that has been association with both neoplastic transformation and the control of cellular growth. Recent studies have reported that p53 also acts as a transcriptional regulator. We have studied transactivational properties of human wild-type and mutant p53 proteins representing 4 major mutational hotspots (codons 141, 175, 248, 273) as well as a double mutant Tyr141/His273 and a p53 with the transcriptional activating region removed (pcDC2). Transactivation by p53 was shown with a p53 concensus binding sequence controlled CAT reporter gene, and activity was assayed after co-transfection of the reporter with either wild-type or mutant p53 expression constructs. Wild-type p53 as well as one mutant p53 [(mutation of arginine to histidine at codon 273 (His 273)], had strong transactivating activity, but all other mutant p53s were inactive in transcriptional activation, including the double mutant Tyr141/His273 suggesting that the Tyr141 mutation was dominant over the His273 mutation in the same protein. Moreover, when mutant p53 (Tyr141, His175, Trp248, or Tyr141/His273) was cotransfected with either wild-type p53 or mutant His273 p53, these mutants inhibited the transactivation of coexpressed wild-type p53. The p53 vector (pcDC2), which contains p53 oligomerization sequences, but not the transactivational domain, markedly inhibited wild-type p53 transactivational activity. Each of the mutant p53s similarly inhibited the transactivation of His273 p53. Therefore, with the exception of His273, each of the other mutant p53 were unable to transactivate and each behaved in a dominant negative fashion.
Hachiya
M
M
Department of Medicine, Cedars-Sinai Medical Center, UCLA School of Medicine 90048.
Chumakov
A
A
Miller
C W
CW
Akashi
M
M
Said
J
J
Koeffler
H P
HP
D001483:Base Sequence; D001665:Binding Sites; D003062:Codon; D016384:Consensus Sequence; D017930:Genes, Reporter; D016158:Genes, p53; D006639:Histidine / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D014158:Transcription, Genetic; D015533:Transcriptional Activation / Q000187:drug effects; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000502:physiology*
D003062:Codon; D016159:Tumor Suppressor Protein p53; D006639:Histidine