The basic carboxy-terminal domain of human p53 is dispensable for both transcriptional regulation and inhibition of tumor cell growth.
To investigate the relevance of the C-terminal domains of the human p53 tumor suppressor gene to its growth suppressive and transcriptional regulatory properties deletion mutants were generated which eliminated 30 (p53 delta 363), 60 (p53 delta 333) and 87 (p53 delta 306) amino acids from the C-terminus of the p53 protein. p53 delta 363 has lost the highly basic tail of the protein (residues 360-386). p53 delta 333 and p53 delta 306 lack the oligomerization domain (residues 320-360); p53 delta 306 has also lost the major nuclear localization signal of p53 (NLSI, residues 316-325). These mutants were assayed for transactivation from two p53 consensus binding sites and for transcriptional repression of two promoter systems in Calu6 lung cancer cells (p53 null). Moreover, their ability to inhibit cell growth in tumor cell lines with a defined p53 status was analysed. Deletion of the oligomerization domain correlated with significant loss of: (a) transactivation from a genomic sequence; (b) transcriptional repression; (c) the ability to inhibit colony formation. An intact NLSI was not a prerequisite for transactivation. p53 delta 363 behaved similarly to wt p53 in all the assays. We established an inducible expression system for p53 delta 363 in a human fibrosarcoma cell line known to be growth-suppressed by wt p53. The induction of p53 delta 363 expression also inhibited cell proliferation albeit to a lesser extent than wt p53. However, p53 delta 363 could upregulate WAF1/CIP1, GADD45 and MDM2 genes. Thus, the basis tail of p53 appears not to be required for the biological functions of the protein assayed.
Pellegata
N S
NS
Department of Microbiology and Molecular Genetics, University of California, Irvine 92717, USA.
Cajot
J F
JF
Stanbridge
E J
EJ
D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects; D002467:Cell Nucleus / Q000737:chemistry; D002471:Cell Transformation, Neoplastic / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic; D016158:Genes, p53; D006801:Humans; D008175:Lung Neoplasms / Q000473:pathology; D008969:Molecular Sequence Data; D009154:Mutation; D010446:Peptide Fragments / Q000737:chemistry*; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology
D010446:Peptide Fragments; D016159:Tumor Suppressor Protein p53