The WT1 gene product stabilizes p53 and inhibits p53-mediated apoptosis.

The Wilms' tumor-suppressor gene product WT1 coimmunoprecipitates with p53 from baby rat kidney (BRK) cells and Wilms' tumor specimens, and expression of WT1 in BRK cells is associated with increased levels of endogenous wild-type p53 protein. To study the effect of WT1 on p53 function, we cotransfected expression constructs into Saos-2 cells, an osteosarcoma cell line without endogenous expression of either gene. Expression of WT1 resulted in increased steady-state levels of p53, attributable to a prolongation in protein half-life, and associated with protection against papillomavirus E6-mediated degradation of p53. This effect mapped to zinc fingers 1 and 2 of WT1 and was not observed with the closely related EGR1 protein. The stabilized p53 demonstrated enhanced binding to its target DNA sequence and increased trans-activation of a promoter containing this RGC site, but reduced transcriptional repression of a TATA-containing promoter lacking this site. Expression of WT1 inhibited p53-mediated apoptosis triggered by UV irradiation or by expression of temperature-sensitive p53 in the wild-type conformation, but did not affect p53-mediated cell cycle arrest. We conclude that WT1 protein can stabilize p53, modulate its trans-activational properties, and inhibit its ability to induce apoptosis. This effect may contribute to the elevated levels of wild-type p53 protein that are observed in Wilms' tumors.

[1]  D. Haber,et al.  WT1 suppresses synthesis of the epidermal growth factor receptor and induces apoptosis. , 1995, The EMBO journal.

[2]  J. Martinou,et al.  Cloning of a bcl-2 homologue by interaction with adenovirus E1B 19K , 1995, Nature.

[3]  K. Miyagawa,et al.  Subnuclear localization of WT1 in splicing or transcription factor domains is regulated by alternative splicing , 1995, Cell.

[4]  V. Metelev,et al.  Induction of apoptosis in uninfected lymphocytes by HIV-1 Tat protein. , 1995, Science.

[5]  Matthew J. Brauer,et al.  Modulation of apoptosis by the widely distributed Bcl-2 homologue Bak , 1995, Nature.

[6]  E. White,et al.  Modulation of p53-mediated transcriptional repression and apoptosis by the adenovirus E1B 19K protein , 1995, Molecular and cellular biology.

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

[8]  N. Nowak,et al.  Anaplastic Wilms' tumour, a subtype displaying poor prognosis, harbours p53 gene mutations , 1994, Nature Genetics.

[9]  M. Sporn,et al.  Repression of the transforming growth factor-beta 1 gene by the Wilms' tumor suppressor WT1 gene product. , 1994, Molecular endocrinology.

[10]  B. Williams,et al.  Mutations of the p53 tumor suppressor gene occur infrequently in Wilms' tumor. , 1994, Cancer research.

[11]  D. Haber,et al.  WT1-mediated growth suppression of Wilms tumor cells expressing a WT1 splicing variant. , 1993, Science.

[12]  S. van den Heuvel,et al.  Distinct roles for cyclin-dependent kinases in cell cycle control. , 1993, Science.

[13]  Xin Lu,et al.  Differential induction of transcriptionally active p53 following UV or lonizing radiation: Defects in chromosome instability syndromes? , 1993, Cell.

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

[15]  M. Scheffner,et al.  The HPV-16 E6 and E6-AP complex functions as a ubiquitin-protein ligase in the ubiquitination of p53 , 1993, Cell.

[16]  X. Xia,et al.  Inhibition of colony-stimulating factor-1 promoter activity by the product of the Wilms' tumor locus. , 1993, The Journal of biological chemistry.

[17]  Z. Wang,et al.  A second transcriptionally active DNA-binding site for the Wilms tumor gene product, WT1. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[18]  S. Capitani,et al.  Human immunodeficiency virus type 1 Tat protein protects lymphoid, epithelial, and neuronal cell lines from death by apoptosis. , 1993, Cancer research.

[19]  David Housman,et al.  WT-1 is required for early kidney development , 1993, Cell.

[20]  C. Thompson,et al.  bcl-x, a bcl-2-related gene that functions as a dominant regulator of apoptotic cell death , 1993, Cell.

[21]  M. Scheffner,et al.  Localization of the E6-AP regions that direct human papillomavirus E6 binding, association with p53, and ubiquitination of associated proteins , 1993, Molecular and cellular biology.

[22]  M. Ewen,et al.  Inhibition of cell proliferation by p107, a relative of the retinoblastoma protein. , 1993, Genes & development.

[23]  V. Sukhatme,et al.  Increased expression of the insulin-like growth factor I receptor gene, IGF1R, in Wilms tumor is correlated with modulation of IGF1R promoter activity by the WT1 Wilms tumor gene product. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[24]  C. Miller,et al.  The p53 activation domain binds the TATA box-binding polypeptide in Holo-TFIID, and a neighboring p53 domain inhibits transcription , 1993, Molecular and cellular biology.

[25]  Amy Bernard,et al.  Physical and functional interaction between WT1 and p53 proteins. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[26]  J. Pipas,et al.  Specific repression of TATA-mediated but not initiator-mediated transcription by wild-type p53 , 1993, Nature.

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

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

[29]  E. White,et al.  Wild-type p53 mediates apoptosis by E1A, which is inhibited by E1B. , 1993, Genes & development.

[30]  S. Lowe,et al.  Stabilization of the p53 tumor suppressor is induced by adenovirus 5 E1A and accompanies apoptosis. , 1993, Genes & development.

[31]  M. Scheffner,et al.  Cloning and expression of the cDNA for E6-AP, a protein that mediates the interaction of the human papillomavirus E6 oncoprotein with p53 , 1993, Molecular and cellular biology.

[32]  John Calvin Reed,et al.  Bcl-2 blocks apoptosis in cells lacking mitochondrial DNA , 1993, Nature.

[33]  G. Zambetti,et al.  Wild-type p53 binds to the TATA-binding protein and represses transcription. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[34]  D. Bonthron,et al.  Human platelet-derived growth factor A chain is transcriptionally repressed by the Wilms tumor suppressor WT1. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[35]  B. Vogelstein,et al.  A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia , 1992, Cell.

[36]  Z. Wang,et al.  The Wilms' tumor gene product, WT1, represses transcription of the platelet-derived growth factor A-chain gene. , 1992, The Journal of biological chemistry.

[37]  N. Lemoine,et al.  Aberrant expression of the tumour suppressor gene p53 is very frequent in Wilms' tumours , 1992, The Journal of pathology.

[38]  T. Crook,et al.  Human papillomavirus E6 proteins bind p53 in vivo and abrogate p53‐mediated repression of transcription. , 1992, The EMBO journal.

[39]  G I Bell,et al.  Repression of the insulin-like growth factor II gene by the Wilms tumor suppressor WT1. , 1992, Science.

[40]  W. Bickmore,et al.  Modulation of DNA binding specificity by alternative splicing of the Wilms tumor wt1 gene transcript. , 1992, Science.

[41]  D. Lane,et al.  p53, guardian of the genome , 1992, Nature.

[42]  P. Sharp,et al.  A dominant mutation in the Wilms tumor gene WT1 cooperates with the viral oncogene E1A in transformation of primary kidney cells. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[43]  A. Levine,et al.  Wild-type p53 mediates positive regulation of gene expression through a specific DNA sequence element. , 1992, Genes & development.

[44]  A. Levine,et al.  The mdm-2 oncogene product forms a complex with the p53 protein and inhibits p53-mediated transactivation , 1992, Cell.

[45]  M. Gossen,et al.  Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[46]  A. Berk,et al.  Inhibition of p53 transactivation required for transformation by adenovirus early 1B protein , 1992, Nature.

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

[48]  L. Donehower,et al.  Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours , 1992, Nature.

[49]  M. Scheffner,et al.  A cellular protein mediates association of p53 with the E6 oncoprotein of human papillomavirus types 16 or 18. , 1991, The EMBO journal.

[50]  M. Yaniv,et al.  Wild-type p53 can down-modulate the activity of various promoters. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[51]  D. Housman,et al.  Alternative splicing and genomic structure of the Wilms tumor gene WT1. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[52]  S. Madden,et al.  Transcriptional repression mediated by the WT1 Wilms tumor gene product. , 1991, Science.

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

[54]  K. Kinzler,et al.  Identification of p53 as a sequence-specific DNA-binding protein , 1991, Science.

[55]  S. Tapscott,et al.  The MCK enhancer contains a p53 responsive element. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[56]  Arnold J. Levine,et al.  The E6 oncoprotein encoded by human papillomavirus types 16 and 18 promotes the degradation of p53 , 1990, Cell.

[57]  T. Curran,et al.  Binding of the Wilms' tumor locus zinc finger protein to the EGR-1 consensus sequence. , 1990, Science.

[58]  B. Vogelstein,et al.  p53 functions as a cell cycle control protein in osteosarcomas , 1990, Molecular and cellular biology.

[59]  O. Halevy,et al.  Conditional inhibition of transformation and of cell proliferation by a temperature-sensitive mutant of p53 , 1990, Cell.

[60]  J. Bard,et al.  The candidate Wilms' tumour gene is involved in genitourinary development , 1990, Nature.

[61]  D. Housman,et al.  An internal deletion within an 11p13 zinc finger gene contributes to the development of Wilms' tumor , 1990, Cell.

[62]  A. Levine,et al.  Association of human papillomavirus types 16 and 18 E6 proteins with p53. , 1990, Science.

[63]  A. Poustka,et al.  Homozygous deletion in Wilms tumours of a zinc-finger gene identified by chromosome jumping , 1990, Nature.

[64]  D. Housman,et al.  Isolation and characterization of a zinc finger polypeptide gene at the human chromosome 11 Wilms' tumor locus , 1990, Cell.

[65]  J. Fargnoli,et al.  Mammalian genes coordinately regulated by growth arrest signals and DNA-damaging agents , 1989, Molecular and cellular biology.

[66]  J. Jenkins,et al.  Ability of p53 and the adenovirus E1b 58-kilodalton protein to form a complex is determined by p53 , 1989, Journal of virology.

[67]  Eileen D. Adamson,et al.  A zinc finger-encoding gene coregulated with c-fos during growth and differentiation, and after cellular depolarization , 1988, Cell.

[68]  H. Koeffler,et al.  Rearrangement of the p53 gene in human osteogenic sarcomas. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[69]  P. Rigby,et al.  High efficiency gene transfer into mammalian cells. , 1984, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[70]  A. Levine,et al.  Adenovirus E1b-58kd tumor antigen and SV40 large tumor antigen are physically associated with the same 54 kd cellular protein in transformed cells , 1982, Cell.

[71]  S. N. Agoff,et al.  Regulation of the human hsp70 promoter by p53. , 1993, Science.