Skip to main content
Log in

A putative protein inhibitor of activated STAT (PIASy) interacts with p53 and inhibits p53-mediated transactivation but not apoptosis

  • Published:
Apoptosis Aims and scope Submit manuscript

Abstract

The p53 protein has recently been reported to be capable of mediating apoptosis through a pathway that is not dependent on its transactivation function. We report here that the PIASy member of the protein inhibitor of activated STAT family inhibited p53's transactivation function without compromising its ability to induce apoptosis of the H1299 nonsmall cell lung carcinoma cell line. The p53 protein bound to PIASy in yeast two-hybrid assays and coprecipitated in complexes with p53 in immunoprecipitates from mammalian cells. PIASy inhibited the DNA-binding activity of p53 in nuclear extracts and blocked the ability of p53 to induce expression of two of its target genes, Bax and p21Waf1/Cip1, in H1299 cells. The block in p53-mediated induction of Bax and p21 was determined to be at the level of transactivation, since PIASy inhibited p53's ability to transactivate a p21/luciferase reporter construct. PIASy did not effect the incidence of apoptosis in H1299 cells upregulated for p53. PIASy appears to regulate p53-mediated functions and may direct p53 into a transactivation-independent mode of apoptosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Martinez J, Georgoff I, Martinez J, Levine AJ. Cellular localization and cell cycle regulation by a temperature-sensitive p53 protein. Genes Develop 1991; 5: 151-159.

    Google Scholar 

  2. Diller L, Kassel J, Nelson CE, et al. p53 functions as a cell cycle control protein in osteosarcomas. Mol Cell Biol 1990; 10: 5772-5781.

    Google Scholar 

  3. Lin D, Shields MT, Ullrich SJ, Apella E, Mercer WE. Growth arrest induced by wild-type p53 protein blocks cells prior to or near the restriction point in late G1 phase. Proc Natl Acad Sci USA 1992; 89: 9210-9214.

    Google Scholar 

  4. Alonigrinstein R, Zanbar I, Alboum I, Goldfinger N, Rotter V. Wild-type p53 functions as a control protein in the differentiation pathway of the B-cell lineage. Oncogene 1993; 8: 3297-3305.

    Google Scholar 

  5. Yin Y, Tainsky MA, Bischoff FZ, Strong LC, Wahl GM. Wild-type p53 restores cell cycle control and inhibits gene amplification in cells with mutant p53 alleles. Cell 1992; 70: 937-948.

    Google Scholar 

  6. Livingstone LR, White A, Sprouse J, Livanos E, Jacks T, Tlsty TD. Altered cell cycle arrest and gene amplification potential accompany loss of wild-type p53. Cell 1992; 70: 923-935.

    Google Scholar 

  7. Dameron KM, Volpert OV, Tainsky MA, Bouck N. Control of angiogenesis in fibroblasts by p53 regulation of thrombospondin-1. Science 1994; 265: 1582-1584.

    Google Scholar 

  8. Stromblad S, Becker JC, Yebra M, Brooks PC, Cheresh DA. Suppression of p53 activity and p21WAF1/cip1 expression by vascular cell integrin alpha V beta 3 during angiogenesis. J Clin Invest 1996; 98: 426-433.

    Google Scholar 

  9. Yonish-Rouach E, Grunwald D, Wilder S, et al. p53-mediated cell death: Relationship to cell cycle control. Mol Cell Biol 1993; 13: 1415-1423.

    Google Scholar 

  10. Debbas M, White E. Wild-type p53 mediates apoptosis by E1A, which is inhibited by E1B. Genes Develop 1993; 7: 546-554.

    Google Scholar 

  11. Clarke AR, Purdie CA, Harrison DJ, et al. Thymocyte apoptosis induced by p53-dependent and independent pathways. Nature 1993; 362: 849-852.

    Google Scholar 

  12. Gottlieb E, Haffner R, Von Ruden T, Wagner EF, Oren M. Down-regulation of wild-type p53 activity interferes with apoptosis of IL-3-dependent hematopoietic cells following IL-3 wilthdrawal. EMBO J 1994; 13: 1368-1374.

    Google Scholar 

  13. Merritt AJ, Potten CS, Kemp CJ, et al. The role of p53 in spontaneous and radiation-induced apoptosis in the gastrointestinal tract of normal and p53-deficient mice. Cancer Res 1994; 54: 614-617.

    Google Scholar 

  14. Hollstein M, Rice K, Greenblatt MS, et al. Database of p53 gene somatic mutations in human tumors and cell lines. Nucl Acids Res 1994; 22: 3551-3555.

    Google Scholar 

  15. Oliner JD, Kinzler KW, Meltzer PS, George DL, Vogelstein B. Amplification of the gene encoding a p53-associated protein in human sarcomas. Nature 1992; 358: 80-83.

    Google Scholar 

  16. Oliner JD, Pietenpol JA, Thiagalingam S, Gyuris J, Kinzler KW, Bogelstein B. Oncoprotein MDM2 conceals the activation domain of the tumour suppressor p53. Nature 1993; 362: 857-860.

    Google Scholar 

  17. Momand J, Zambetti GP, Olson DC, George D, Levine AJ. The mdm-2 oncogene product forms a complex with the p53 protein and inhibits p53-mediated transactivation. Cell 1992; 69: 1237-1245.

    Google Scholar 

  18. Chen, C-Y, Oliner JD, Zhan Q, Fornace AJ Jr., Vogelstein B, Kastan MB. Interactions between p53 and MDM2 in a mammalian cell cycle checkpoint pathway. Proc Natl Acad Sci USA 1994; 91: 2684-2688.

    Google Scholar 

  19. Levine AJ. p53, the cellular gatekeeper for growth and division. Cell 1997; 88: 323-331.

    Google Scholar 

  20. Ko LJ, Prives C. p53: Puzzle and paradigm. Genes Develop 1996; 10: 1054-1072.

    Google Scholar 

  21. Prives C, Hall P. The p53 pathway. J of Pathology 1999; 187: 112-126.

    Google Scholar 

  22. Giaccia AJ, Kastan M. The complexity of p53 modulation: Emerging patterns from divergent signals. Genes and Development 1998; 12: 2973-2983.

    Google Scholar 

  23. El-Deiry WS. Regulation of p53 downstream genes. Seminars in Cancer Biology 1998; 8: 345-357.

    Google Scholar 

  24. Tanaka H, Arakawa H, Yamaguchi T, et al. A ribonucleotide reductase gene involved in a p53-dependent cell-cycle checkpoint for DNA damage. Nature 2000; 404: 42-49.

    Google Scholar 

  25. Del Sal G, Ruaro EM, Utrera R, Cole C, Levine AJ, Schneider C. Gas1-induced growth suppression requires a transactivation-independent p53 function. Molecular and Cellular Biology 1995; 15: 7152-7160.

    Google Scholar 

  26. Theis S, Atz J, Mueller-Lantzsch N, Roemer K. A function in apoptosis other than transactivation inherent in the NH2-terminal domain of p53. Int J Cancer 1997; 71: 858-866.

    Google Scholar 

  27. Atz J, Wagner P, Roemer K. Function, oligomerization, and conformation of tumor-associated p53 proteins with mutated C-terminus. J of Cellular Biochemistry 2000; 76: 572-584.

    Google Scholar 

  28. Gao C, Tsuchida N. Activation of caspases in p53-induced transactivation-independent apoptosis. Jpn J Cancer Res 1999; 90: 180-187.

    Google Scholar 

  29. Takahashi T, Carbone D, Takahashi T, et al. Wild-type but not mutant p53 suppresses the growth of human lung cancer cells bearing multiple genetic lesions. Cancer Res 1992; 52: 2340-2343.

    Google Scholar 

  30. Ito H, Fukada Y, Murata K, Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol 1983; 153: 163-168.

    Google Scholar 

  31. Breeden L, Naysmyth K. Regulation of the yeast HO Gene. Cold Harbor Symposium Quant Biol 1985; 50: 643-650.

    Google Scholar 

  32. Ward A. Single-step purification of shuttle vectors from yeast for high frequency back-transformation into E. coli. Nucl Acids Res 1990; 18: 5319.

    Google Scholar 

  33. Maxwell SA, Acosta SA, Tombusch K, Davis GE. Expression of bax, bcl-2, waf-1, and PCNA gene products in an immortalized human endothelial cell line undergoing p53-mediated apoptosis. Apoptosis 1997; 2:442-454.

    Google Scholar 

  34. Dignam JD, Lebovitz RM, Roeder RG. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Research 1983; 11: 1475-1489.

    Google Scholar 

  35. Unger T, Sionov RV, Moallem E, et al. Mutations in serines 15 and 20 of human p53 impair its apoptotic ability. Oncogene 1999; 18: 3205-3212.

    Google Scholar 

  36. Maxwell SA, Davis GE. Molecular and biological characterization of an ECV-304-derived cell line resistant to p53-mediated apoptosis. Apoptosis 2000; 5, 277-288.

    Google Scholar 

  37. Liu B, Liao J, Rao X, et al. Inhibition of Stat1-mediated gene activation by PIAS1. Proc Natl Acad Sci USA 1998; 95: 10626-10631.

    Google Scholar 

  38. Simbulan-Rosenthal C, Rosenthal D, Iyer S, Boulares H, Smulson M. Transient poly(ADP-ribosyl)ation of nuclear protein and role of poly(ADP-ribose) polymerase in the early stages of apoptosis. J Biol Chem 1998; 273: 13703-13712.

    Google Scholar 

  39. Boulares AH, Yakovlev AG, Ivanoca V, et al. Role of poly(ADPribose) polymerase (PARP) cleavage in apoptosis. Caspase 3-resistant PARP mutant increases rates of apoptosis in transfected cells. J Biol Chem 1999; 274: 22932-22940.

    Google Scholar 

  40. Germain M, Affar ED, D'Amours D, Dixit VM, Salvesen GS, Poirier GG. Cleavage of automodified poly(ADP-ribose) polymerase during apoptosis. Evidence for involvement of caspase-7. J Biol Chem 1999; 274: 28379-28384.

    Google Scholar 

  41. Salvesen GS Dixit VM. Caspases: Intracellular signaling by proteolysis. Cell 1997; 91: 443-446.

    Google Scholar 

  42. Hoey T. A new player in cell death. Science 1997; 278: 1578-1579.

    Google Scholar 

  43. Sierra-Honigman MR, Nath AK, Murakami C, et al. Biological action of leptin as an angiogenic factor. Science 1998; 281: 1683-1686.

    Google Scholar 

  44. Ihle JN. STATs: Signal transducers and activators of transcripton. Cell 1996; 84: 331-334.

    Google Scholar 

  45. Rayanade RJ, Ndubuisi MI, Etlinger JD, Sehgal PB. Regulation of IL-6 signaling by p53: Stat3-and Stat5-masking in p53-Val135-containing human hepatoma Hep3B cell lines. J Immunol 1998; 161: 325-333.

    Google Scholar 

  46. Ritchie A, Braun SE, He J, Broxmeyer HE. Thrombopoietininduced conformational change in p53 lies downstream of the p44/p42 mitogen activated protein kinase cascade in the human growth factor-dependent cell line M07e. Oncogene 1999; 18: 1465-1477.

    Google Scholar 

  47. Fritsche M, Mundt M, Merkle C, Jahne R, Groner B. p53 suppresses cytokine induced, Stat5 mediated activation of transcription. Mol Cell Endocrinology 1998; 143: 143-154.

    Google Scholar 

  48. Garcia R, Jove R. Activation of STAT transcription factors in oncogenic tyrosine kinase signaling. J of Biomedical Science 1998; 5: 79-85.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nelson, V., Davis, G.E. & Maxwell, S.A. A putative protein inhibitor of activated STAT (PIASy) interacts with p53 and inhibits p53-mediated transactivation but not apoptosis. Apoptosis 6, 221–234 (2001). https://doi.org/10.1023/A:1011392811628

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1011392811628

Navigation