Genetic selection of intragenic suppressor mutations that reverse the effect of common p53 cancer mutations

Several lines of evidence suggest that the presence of the wild‐type tumor suppressor gene p53 in human cancers correlates well with successful anti‐cancer therapy. Restoration of wild‐type p53 function to cancer cells that have lost it might therefore improve treatment outcomes. Using a systematic yeast genetic approach, we selected second‐site suppressor mutations that can overcome the deleterious effects of common p53 cancer mutations in human cells. We identified several suppressor mutations for the V143A, G245S and R249S cancer mutations. The beneficial effects of these suppressor mutations were demonstrated using mammalian reporter gene and apoptosis assays. Further experiments showed that these suppressor mutations could override additional p53 cancer mutations. The mechanisms of such suppressor mutations can be elucidated by structural studies, ultimately leading to a framework for the discovery of small molecules able to stabilize p53 mutants.

[1]  D. Lane,et al.  Activation of the cryptic DNA binding function of mutant forms of p53. , 1993, Nucleic acids research.

[2]  L. Donehower,et al.  The tumor suppressor p53. , 1993, Biochimica et biophysica acta.

[3]  K. Kinzler,et al.  Oncogenic forms of p53 inhibit p53-regulated gene expression. , 1992, Science.

[4]  D. Shortle,et al.  Genetic analysis of staphylococcal nuclease: identification of three intragenic "global" suppressors of nuclease-minus mutations. , 1985, Genetics.

[5]  R. Hruban,et al.  p 53 Mutations in Pancreatic Carcinoma and Evidence of Common Involvement of Homocopolymer Tracts in DNA Microdeletions 1 , 2006 .

[6]  C. Harris,et al.  Mutations in the p53 tumor suppressor gene: clues to cancer etiology and molecular pathogenesis. , 1994, Cancer research.

[7]  C. Prives,et al.  p53: puzzle and paradigm. , 1996, Genes & development.

[8]  M. Vidal,et al.  Dominant-negative p53 mutations selected in yeast hit cancer hot spots. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[9]  T. Halazonetis,et al.  Structure–based rescue of common tumor–derived p53 mutants , 1996, Nature Medicine.

[10]  W. Hong,et al.  Retrovirus–mediated wild–type P53 gene transfer to tumors of patients with lung cancer. , 1996, Nature Medicine.

[11]  M. Vidal,et al.  Reverse two-hybrid and one-hybrid systems to detect dissociation of protein-protein and DNA-protein interactions. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[12]  D. Abraham,et al.  Allosteric modifiers of hemoglobin: 2-[4-[[(3,5-disubstituted anilino)carbonyl]methyl]phenoxy]-2-methylpropionic acid derivatives that lower the oxygen affinity of hemoglobin in red cell suspensions, in whole blood, and in vivo in rats. , 1992, Biochemistry.

[13]  R. Sikorski,et al.  A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. , 1989, Genetics.

[14]  J. Gibbs,et al.  Pharmaceutical research in molecular oncology , 1994, Cell.

[15]  B. Groner,et al.  Restoration of the growth suppression function of mutant p53 by a synthetic peptide derived from the p53 C-terminal domain , 1997, Nature Medicine.

[16]  A. Neri,et al.  Microinjection of monoclonal antibody PAb421 into human SW480 colorectal carcinoma cells restores the transcription activation function to mutant p53. , 1995, Cancer research.

[17]  D. Fisher.  Apoptosis in cancer therapy: Crossing the threshold , 1994, Cell.

[18]  M. Stratton,et al.  Mutation of the p53 gene in human soft tissue sarcomas: association with abnormalities of the RB1 gene. , 1990, Oncogene.

[19]  R. Hruban,et al.  p53 mutations in pancreatic carcinoma and evidence of common involvement of homocopolymer tracts in DNA microdeletions. , 1994, Cancer research.

[20]  P. Jeffrey,et al.  Crystal structure of a p53 tumor suppressor-DNA complex: understanding tumorigenic mutations. , 1994, Science.

[21]  Thierry Soussi,et al.  Somatic point mutations in the p53 gene of human tumors and cell lines: updated compilation , 1996, Nucleic Acids Res..

[22]  Junying Yuan,et al.  Induction of apoptosis in fibroblasts by IL-1β-converting enzyme, a mammalian homolog of the C. elegans cell death gene ced-3 , 1993, Cell.

[23]  E. Baxter,et al.  p53 and tumour viruses: catching the guardian off-guard. , 1997, Trends in microbiology.

[24]  D. Housman,et al.  p53 status and the efficacy of cancer therapy in vivo. , 1994, Science.

[25]  B. Vogelstein,et al.  p53 mutations in human cancers. , 1991, Science.

[26]  D. Lane,et al.  The dominating effect of mutant p53 , 1995, Nature Genetics.

[27]  O. Halevy,et al.  p53 Mutations: Gains or losses? , 1991, Journal of cellular biochemistry.

[28]  E. Shaulian,et al.  Induction of apoptosis in HeLa cells by trans-activation-deficient p53. , 1995, Genes & development.

[29]  E. White,et al.  Life, death, and the pursuit of apoptosis. , 1996, Genes & development.

[30]  T. Halazonetis,et al.  Conformational shifts propagate from the oligomerization domain of p53 to its tetrameric DNA binding domain and restore DNA binding to select p53 mutants. , 1993, The EMBO journal.

[31]  J. Kovarik,et al.  p53 derived from human tumour cell lines and containing distinct point mutations can be activated to bind its consensus target sequence. , 1995, Oncogene.

[32]  K. Kinzler,et al.  Definition of a consensus binding site for p53 , 1992, Nature Genetics.

[33]  Scott W. Lowe,et al.  p53 is required for radiation-induced apoptosis in mouse thymocytes , 1993, Nature.

[34]  Neal F. Cariello,et al.  Databases and software for the analysis of mutations in the human p53 gene, human hprt gene and both the lacI and lacZ gene in transgenic rodents , 1998, Nucleic Acids Res..

[35]  K. Kinzler,et al.  Cancer therapy meets p53. , 1994, The New England journal of medicine.

[36]  M. Oren,et al.  p53 in growth control and neoplasia. , 1996, Biochimica et biophysica acta.

[37]  K. Kinzler,et al.  Sequence-specific transcriptional activation is essential for growth suppression by p53. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[38]  M. Oren,et al.  Biochemical properties and biological effects of p53. , 1995, Current opinion in genetics & development.

[39]  C. Gorman,et al.  Transient production of proteins using an adenovirus transformed cell line , 1990 .

[40]  J. Milner.  DNA damage, p53 and anticancer therapies , 1995, Nature Medicine.

[41]  D A Scudiero,et al.  An abnormality in the p53 pathway following gamma-irradiation in many wild-type p53 human melanoma lines. , 1996, Cancer research.

[42]  Scott A. Peterson,et al.  Inhibiting transthyretin amyloid fibril formation via protein stabilization. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[43]  M. Oren,et al.  p53; from inductive signal to cellular effect. , 1997, Current opinion in genetics & development.

[44]  L. Holmberg,et al.  Complete sequencing of the p53 gene provides prognostic information in breast cancer patients, particularly in relation to adjuvant systemic therapy and radiotherapy , 1995, Nature Medicine.

[45]  D. Lane,et al.  Small peptides activate the latent sequence-specific DNA binding function of p53 , 1995, Cell.

[46]  J. Trent,et al.  WAF1, a potential mediator of p53 tumor suppression , 1993, Cell.

[47]  D. Housman,et al.  p53-dependent apoptosis modulates the cytotoxicity of anticancer agents , 1993, Cell.

[48]  S. Lowe,et al.  Cancer therapy and p53. , 1995, Current opinion in oncology.

[49]  L. Donehower,et al.  The tumore suppressor p53 , 1993 .

[50]  A. Neri,et al.  Restoration of the transcription activation function to mutant p53 in human cancer cells. , 1996, Oncogene.

[51]  R. Parker,et al.  A rapid method for localized mutagenesis of yeast genes , 1992, Yeast.

[52]  R. Iggo,et al.  Mammalian p53 can function as a transcription factor in yeast. , 1992, Nucleic acids research.

[53]  J. Freeman,et al.  Mutation of conserved domain II alters the sequence specificity of DNA binding by the p53 protein. , 1994, The EMBO journal.

[54]  R. Saiki,et al.  A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. , 1988, Nucleic acids research.

[55]  R. Mirimanoff,et al.  Clinical implications of the p53 tumor-suppressor gene. , 1994, The New England journal of medicine.

[56]  Thierry Soussi,et al.  TP53 tumor suppressor gene: A model for investigating human mutagenesis , 1992, Genes, chromosomes & cancer.

[57]  Bert Vogelstein,et al.  p53 function and dysfunction , 1992, Cell.

[58]  M. Karin,et al.  p53-Dependent apoptosis in the absence of transcriptional activation of p53-target genes , 1994, Nature.

[59]  C. Purdie,et al.  Thymocyte apoptosis induced by p53-dependent and independent pathways , 1993, Nature.

[60]  B. Vogelstein,et al.  Suppression of human colorectal carcinoma cell growth by wild-type p53. , 1990, Science.

[61]  Y. Kassir,et al.  A simple and highly efficient procedure for rescuing autonomous plasmids from yeast. , 1992, Nucleic acids research.

[62]  Neal F. Cariello,et al.  Databases and software for the analysis of mutations in the human p53 gene, the human hprt gene and the lacZ gene in transgenic rodents , 1996, Nucleic Acids Res..

[63]  C. Harris,et al.  Structure and function of the p53 tumor suppressor gene: clues for rational cancer therapeutic strategies. , 1996, Journal of the National Cancer Institute.

[64]  A. Levine.  p53, the Cellular Gatekeeper for Growth and Division , 1997, Cell.