The 273rd codon mutants of p53 show growth modulation activities not correlated with p53-specific transactivation activity.
Cancer-related mutations of the p53 tumor suppressor gene are clustered in the four so-called 'hot spots', codons 175, 248, 273 and 281/282. By using recombination PCR in vitro mutagenesis, we introduced point mutations into the codon 273 of wild-type (wt) p53 (pC53-SN3) from Arg to His (pC53-273H [273H]), Asp (273D), Pro (273P), Lys (273K), Leu (273L) or Thr (273T), and compared their biological and biochemical activities with wt p53 and cancer-derived 175H, 248W and 273H/309S. Among them, 273H/309S, 273H and 273D as well as wt p53 transactivated the chloramphenicol acetyltransferase (CAT) gene placed downstream of the p53 binding consensus, while none of the other mutants including 273L did. Transcriptions from human c-fos and rat PCNA promoters were suppressed by wt p53 and 273D, while they were enhanced variously by all other mutants in Saos-2 and/or NIH3T3 cells. On the other hand, growth of human squamous carcinoma cell lines measured by the plating efficiency of G418-resistant colonies was enhanced by transfection of 175H, 248W, 273H/309S and 273P, while suppressed by not only wt p53, 273D and 273H but also 273L. Thus, 273H/309S enhanced cell growth in spite of its p53-specific transactivation activity, while 273L suppressed cell growth in spite of its complete loss of the p53-specific transactivation. We concluded that the sequence-specific transactivation of p53 is not always correlated with its growth inhibitory activity.
Kawamura
M
M
Department of Molecular Biology, Cancer Research Institute, Sapporo Medical University School of Medicine, Japan.
Yamashita
T
T
Segawa
K
K
Kaneuchi
M
M
Shindoh
M
M
Fujinaga
K
K
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002294:Carcinoma, Squamous Cell / Q000473:pathology; D002455:Cell Division / Q000235:genetics*; D002471:Cell Transformation, Neoplastic; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D003062:Codon / Q000235:genetics / Q000502:physiology*; D017930:Genes, Reporter; D016158:Genes, p53 / Q000235:genetics / Q000502:physiology*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016296:Mutagenesis; D017354:Point Mutation / Q000235:genetics / Q000502:physiology*; D051381:Rats; D015533:Transcriptional Activation / Q000502:physiology*; D014162:Transfection; D014407:Tumor Cells, Cultured; D014410:Tumor Stem Cell Assay
D003062:Codon; D015500:Chloramphenicol O-Acetyltransferase