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Understanding the Effects of Cancer-Associated Mutations in the Tumor Suppressor Protein p53: Structural Consequences of Mutations and Possible Ways of Rescuing Oncogenic Mutants

  • Andreas C. Joerger
  • Assaf Friedler
  • Alan R. Fersht
Part of the Protein Reviews book series (PRON, volume 6)

Abstract

The tumor suppressor protein p53 is a key control in the cell cycle and plays a crucial role in the prevention of cancer development. It is mutated in approximately half of all human cancers and has, therefore, become an important target for the development of novel cancer therapies. Here, we review the structure of the protein, the effects of mutation and how they may be reversed. p53 has a highly complex domain organization consisting of structured regions combined with largely unstructured domains. Most cancer-associated mutations are located in the DNA-binding core domain of the protein. The molecular basis for the detrimental effect of these mutations has been elucidated by structural and biophysical studies. Whereas some mutations affect residues that make direct contact with target DNA, others induce structural perturbations that reduce the thermodynamic stability of the protein. Because p53 core domain is only marginally stable above body temperature, many cancer mutations not only induce local conformational changes but also cause global unfolding of the core domain under physiologic conditions. Novel therapeutic strategies aim, therefore, to develop chemical chaperones that help refold p53 mutants to their correct native structure. Valuable lessons can be learned from studies on so-called second-site suppressor mutations that reverse the effects of cancer mutations. These may provide a basis for the rational design of novel therapeutics.

Keywords

Core Domain Chemical Chaperone Heteronuclear Single Quantum Correlation Tetramerization Domain Color Insert 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Aguilar, F., Hussain, S. P. and Cerutti, P. (1993). Aflatoxin B1 induces the transversion of G → T in codon 249 of the p53 tumor suppressor gene in human hepatocytes. Proc. Natl. Acad. Sci. USA. 90: 8586–8590.PubMedCrossRefGoogle Scholar
  2. Baroni, T. E., Wang, T., Qian, H., Dearth, L. R., Truong, L. N., Zeng, J., Denes, A. E., Chen, S. W. and Brachmann, R. K. (2004). A global suppressor motif for p53 cancer mutants. Proc. Natl. Acad. Sci. USA 101: 4930–4935.PubMedCrossRefGoogle Scholar
  3. Brachmann, R. K., Yu, K., Eby, Y., Pavletich, N. P. and Boeke, J. D. (1998). Genetic selection of intragenic suppressor mutations that reverse the effect of common p53 cancer mutations. EMBO J. 17: 1847–1859.PubMedCrossRefGoogle Scholar
  4. Buchhop, S., Gibson, M. K., Wang, X. W., Wagner, P., Sturzbecher, H. W. and Harris, C. C. (1997). Interaction of p53 with the human Rad51 protein. Nucleic Acids Res. 25: 3868–74.PubMedCrossRefGoogle Scholar
  5. Bullock, A. N., Henckel, J., DeDecker, B. S., Johnson, C. M., Nikolova, P. V., Proctor, M. R., Lane, D. P. and Fersht, A. R. (1997). Thermodynamic stability of wild-type and mutant p53 core domain. Proc. Natl. Acad. Sci. USA 94: 14338–14342.PubMedCrossRefGoogle Scholar
  6. Bullock, A. N., Henckel, J. and Fersht, A. R. (2000). Quantitative analysis of residual folding and DNA binding in mutant p53 core domain: definition of mutant states for rescue in cancer therapy. Oncogene 19: 1245–1256.PubMedCrossRefGoogle Scholar
  7. Bullock, A. N. and Fersht, A. R. (2001). Rescuing the function of mutant p53. Nat. Cancer Rev. 1: 68–76.CrossRefGoogle Scholar
  8. Bykov, V. J., Issaeva, N., Shilov, A., Hultcrantz, M., Pugacheva, E., Chumakov, P., Bergman, J., Wiman, K. G. and Selivanova, G. (2002). Restoration of the tumor suppressor function to mutant p53 by a low-molecular-weight compound. Nat. Med. 8: 282–288.PubMedCrossRefGoogle Scholar
  9. Bykov, V. J., Selivanova, G. and Wiman, K. G. (2003). Small molecules that reactivate mutant p53. Eur. J. Cancer 39: 1828–1834.PubMedCrossRefGoogle Scholar
  10. Cho, Y., Gorina, S., Jeffrey, P. D. and Pavletich, N. P. (1994). Crystal structure of a p53 tumor suppressor-DNA complex: understanding tumorigenic mutations. Science 265: 346–355.PubMedCrossRefGoogle Scholar
  11. Clore, G. M., Ernst, J., Clubb, R., Omichinski, J. G., Kennedy, W. M., Sakaguchi, K., Appella, E. and Gronenborn, A. M. (1995). Refined solution structure of the oligomerization domain of the tumour suppressor p53. Nat. Struct. Biol. 2: 321–333.PubMedCrossRefGoogle Scholar
  12. Clore, G. M., Omichinski, J. G., Sakaguchi, K., Zambrano, N., Sakamoto, H., Appella, E. and Gronenborn, A. M. (1994). High-resolution structure of the oligomerization domain of p53 by multidimensional NMR. Science 265: 386–391.PubMedCrossRefGoogle Scholar
  13. Courtois, S., de Fromentel, C. C. and Hainaut, P. (2004). p53 protein variants: structural and functional similarities with p63 and p73 isoforms. Oncogene 23: 631–638.PubMedCrossRefGoogle Scholar
  14. Derbyshire, D. J., Basu, B. P., Serpell, L. C., Joo, W. S., Date, T., Iwabuchi, K. and Doherty, A. J. (2002). Crystal structure of human 53BP1 BRCT domains bound to p53 tumour suppressor. EMBO J. 21: 3863–3872.PubMedCrossRefGoogle Scholar
  15. DiTullio, R. A., Jr., Mochan, T. A., Venere, M., Bartkova, J., Sehested, M., Bartek, J. and Halazonetis, T. D. (2002). 53BP1 functions in an ATM-dependent checkpoint pathway that is constitutively activated in human cancer. Nat. Cell Biol. 4:998–1002.PubMedCrossRefGoogle Scholar
  16. Dobson, C. M. (1999). Protein misfolding, evolution and disease. Trends Biochem. Sci. 24: 329–332.PubMedCrossRefGoogle Scholar
  17. Fan, J. Q., Ishii, S., Asano, N. and Suzuki, Y. (1999). Accelerated transport and maturation of lysosomal alpha-galactosidase A in Fabry lymphoblasts by an enzyme inhibitor. Nat. Med. 5: 112–115.PubMedCrossRefGoogle Scholar
  18. Fan, J. Q. (2003). A contradictory treatment for lysosomal storage disorders: inhibitors enhance mutant enzyme activity. Trends Pharmacol. Sci. 24: 355–360.PubMedCrossRefGoogle Scholar
  19. Foster, B. A., Coffey, H. A., Morin, M. J. and Rastinejad, F. (1999). Pharmacological rescue of mutant p53 conformation and function. Science 286: 2507–2510.PubMedCrossRefGoogle Scholar
  20. Friedler, A., Hansson, L. O., Veprintsev, D. B., Freund, S. M., Rippin, T. M., Nikolova, P. V., Proctor, M. R., Rüdiger, S. and Fersht, A. R. (2002). A peptide that binds and stabilizes p53 core domain: chaperone strategy for rescue of oncogenic mutants. Proc. Natl. Acad. Sci. USA 99: 937–942.PubMedCrossRefGoogle Scholar
  21. Friedler, A., Veprintsev, D. B., Hansson, L. O. and Fersht, A. R. (2003). Kinetic instability of p53 core domain mutants: implications for rescue by small molecules. J. Biol. Chem. 278: 24108–24112.PubMedCrossRefGoogle Scholar
  22. Friedler, A., DeDecker, B. S., Freund, S. M., Blair, C., Rüdiger, S. and Fersht, A. R. (2004). Structural distortion of p53 by the mutation R249S and its rescue by a designed peptide: implications for “mutant conformation.” J. Mol. Biol. 336: 187–196.PubMedCrossRefGoogle Scholar
  23. Friedler, A., Veprintsev, D. B., Freund, S. M., von Glos, K. I. and Fersht, A. R. (2005a). Modulation of binding of DNA to the C-terminal domain of p53 by acetylation. Structure 13: 629–36.PubMedCrossRefGoogle Scholar
  24. Friedler, A., Veprintsev, D. B., Rutherford, T., von Glos, K. I. and Fersht, A. R. (2005b). Binding of Rad51 and other peptide sequences to a promiscuous, highly electrostatic, binding site in p53. J. Biol. Chem. 280: 8051–8059.PubMedCrossRefGoogle Scholar
  25. Gorina, S. and Pavletich, N. P. (1996). Structure of the p53 tumor suppressor bound to the ankyrin and SH3 domains of 53BP2. Science 274: 1001–1005.PubMedCrossRefGoogle Scholar
  26. Grossman, S. R. (2001). p300/CBP/p53 interaction and regulation of the p53 response. Eur. J. Biochem. 268: 2773–2778.PubMedCrossRefGoogle Scholar
  27. Hainaut, P. and Hollstein, M. (2000). p53 and human cancer: the first ten thousand mutations. Adv. Cancer Res. 77: 81–137.PubMedGoogle Scholar
  28. Issaeva, N., Friedler, A., Bozko, P., Wiman, K. G., Fersht, A. R. and Selivanova, G. (2003). Rescue of mutants of the tumor suppressor p53 in cancer cells by a designed peptide. Proc. Natl. Acad. Sci. USA 100: 13303–13307.PubMedCrossRefGoogle Scholar
  29. Issaeva, N., Bozko, P., Enge, M., Protopopova, M., Verhoef, L. G., Masucci, M., Pramanik, A. and Selivanova, G. (2004). Small molecule RITA binds to p53, blocks p53-HDM-2 interaction and activates p53 function in tumors. Nat. Med. 10:1321–1328.PubMedCrossRefGoogle Scholar
  30. Jeffrey, P. D., Gorina, S. and Pavletich, N. P. (1995). Crystal structure of the tetramerization domain of the p53 tumor suppressor at 1.7 angstroms. Science 267: 1498–1502.PubMedCrossRefGoogle Scholar
  31. Joerger, A. C., Allen, M. D. and Fersht, A. R. (2004). Crystal structure of a superstable mutant of human p53 core domain. Insights into the mechanism of rescuing oncogenic mutations. J. Biol. Chem. 279: 1291–1296.PubMedCrossRefGoogle Scholar
  32. Joerger, A. C., Ang, H. C., Veprintsev, D. B., Blair, C. M. and Fersht, A. R. (2005). Structures of p53 cancer mutants and mechanism of rescue by second-site suppressor mutations. J. Biol. Chem. 280: 16030–16037.PubMedCrossRefGoogle Scholar
  33. Joo, W. S., Jeffrey, P. D., Cantor, S. B., Finnin, M. S., Livingston, D. M. and Pavletich, N. P. (2002). Structure of the 53BP1 BRCT region bound to p53 and its comparison to the Brca1 BRCT structure. Genes Dev. 16: 583–593.PubMedCrossRefGoogle Scholar
  34. Kraulis, P. J. (1991). MOLSCRIPT: a program to produce both detailed and schematic plots of protein structures. J. Appl. Crystallogr. 24: 946–950.CrossRefGoogle Scholar
  35. Kussie, P. H., Gorina, S., Marechal, V., Elenbaas, B., Moreau, J., Levine, A. J. and Pavletich, N. P. (1996). Structure of the MDM2 oncoprotein bound to the p53 tumor suppressor transactivation domain. Science 274: 948–953.PubMedCrossRefGoogle Scholar
  36. Lane, D. P. and Lain, S. (2002). Therapeutic exploitation of the p53 pathway. Trends Mol. Med. 8: S38–S42.PubMedCrossRefGoogle Scholar
  37. Lane, D. P. and Hupp, T. R. (2003). Drug discovery and p53. Drug Discov. Today 8: 347–355.PubMedCrossRefGoogle Scholar
  38. Linke, S. P., Sengupta, S., Khabie, N., Jeffries, B. A., Buchhop, S., Miska, S., Henning, W., Pedeux, R., Wang, X. W., Hofseth, L. J., Yang, Q., Garfield, S. H., Sturzbecher, H. W. and Harris, C. C. (2003). p53 interacts with hRAD51 and hRAD54, and directly modulates homologous recombination. Cancer Res. 63: 2596–605.PubMedGoogle Scholar
  39. Matsumura, I. and Ellington, A. D. (1999). In vitro evolution of thermostable p53 variants. Protein Sci. 8: 731–740.PubMedGoogle Scholar
  40. Merritt, E. A. and Bacon, D. J. (1997). Raster3D: Photorealistic molecular graphics. Methods Enzymol. 277: 505–524.CrossRefPubMedGoogle Scholar
  41. Michael, D. and Oren, M. (2002). The p53 and Mdm2 families in cancer. Curr. Opin. Genet. Dev. 12: 53–59.PubMedCrossRefGoogle Scholar
  42. Michael, D. and Oren, M. (2003). The p53-Mdm2 module and the ubiquitin system. Semin. Cancer Biol. 13: 49–58.PubMedCrossRefGoogle Scholar
  43. Momand, J., Wu, H. H. and Dasgupta, G. (2000). MDM2—master regulator of the p53 tumor suppressor protein. Gene 242: 15–29.PubMedCrossRefGoogle Scholar
  44. Morello, J. P., Petaja-Repo, U. E., Bichet, D. G. and Bouvier, M. (2000). Pharmacological chaperones: a new twist on receptor folding. Trends Pharmacol. Sci. 21: 466–469.PubMedCrossRefGoogle Scholar
  45. Müller-Tiemann, B. F., Halazonetis, T. D. and Elting, J. J. (1998). Identification of an additional negative regulatory region for p53 sequence-specific DNA binding. Proc. Natl. Acad. Sci. USA 95: 6079–6084.PubMedCrossRefGoogle Scholar
  46. Nikolova, P. V., Henckel, J., Lane, D. P. and Fersht, A. R. (1998). Semirational design of active tumor suppressor p53 DNA binding domain with enhanced stability. Proc. Natl. Acad. Sci. USA 95: 14675–14680.PubMedCrossRefGoogle Scholar
  47. Nikolova, P. V., Wong, K. B., DeDecker, B., Henckel, J. and Fersht, A. R. (2000). Mechanism of rescue of common p53 cancer mutations by second-site suppressor mutations. EMBO J. 19: 370–378.PubMedCrossRefGoogle Scholar
  48. Olivier, M., Eeles, R., Hollstein, M., Khan, M. A., Harris, C. C. and Hainaut, P. (2002). The IARC TP53 database: new online mutation analysis and recommendations to users. Hum. Mutat. 19: 607–614.PubMedCrossRefGoogle Scholar
  49. Peng, Y., Li, C., Chen, L., Sebti, S. and Chen, J. (2003). Rescue of mutant p53 transcription function by ellipticine. Oncogene 22: 4478–4487.PubMedCrossRefGoogle Scholar
  50. Prives, C. and Manley, J. L. (2001). Why is p53 acetylated? Cell 107: 815–818.PubMedCrossRefGoogle Scholar
  51. Resnick, M. A. and Inga, A. (2003). Functional mutants of the sequence-specific transcription factor p53 and implications for master genes of diversity. Proc. Natl. Acad. Sci. USA 100: 9934–9939.PubMedCrossRefGoogle Scholar
  52. Rippin, T. M., Bykov, V. J., Freund, S. M., Selivanova, G., Wiman, K. G. and Fersht, A. R. (2002). Characterization of the p53-rescue drug CP-31398 in vitro and in living cells. Oncogene 21: 2119–2129.PubMedCrossRefGoogle Scholar
  53. Rustandi, R. R., Baldisseri, D. M. and Weber, D. J. (2000). Structure of the negative regulatory domain of p53 bound to S100B(betabeta). Nat. Struct. Biol. 7: 570–574.PubMedCrossRefGoogle Scholar
  54. Ryan, K. M., Phillips, A. C. and Vousden, K. H. (2001). Regulation and function of the p53 tumor suppressor protein. Curr. Opin. Cell Biol. 13: 332–337.PubMedCrossRefGoogle Scholar
  55. Samuels-Lev, Y., O’Connor, D. J., Bergamaschi, D., Trigiante, G., Hsieh, J. K., Zhong, S., Campargue, I., Naumovski, L., Crook, T. and Lu, X. (2001). ASPP proteins specifically stimulate the apoptotic function of p53. Mol. Cell 8: 781–794.PubMedCrossRefGoogle Scholar
  56. Selivanova, G., Iotsova, V., Okan, I., Fritsche, M., Strom, M., Groner, B., Grafstrom, R. C. and Wiman, K. G. (1997). Restoration of the growth suppression function of mutant p53 by a synthetic peptide derived from the p53 C-terminal domain. Nat. Med. 3: 632–638.PubMedCrossRefGoogle Scholar
  57. Selivanova, G., Kawasaki, T., Ryabchenko, L. and Wiman, K. G. (1998). Reactivation of mutant p53: a new strategy for cancer therapy. Semin. Cancer Biol. 8: 369–378.PubMedCrossRefGoogle Scholar
  58. Selivanova, G., Ryabchenko, L., Jansson, E., Iotsova, V. and Wiman, K. G. (1999). Reactivation of mutant p53 through interaction of a C-terminal peptide with the core domain. Mol. Cell Biol. 19: 3395–3402.PubMedGoogle Scholar
  59. Shaulsky, G., Goldfinger, N., Ben-Zeev, A. and Rotter, V. (1990). Nuclear accumulation of p53 protein is mediated by several nuclear localization signals and plays a role in tumorigenesis. Mol. Cell. Biol. 10: 6565–6577.PubMedGoogle Scholar
  60. Stommel, J. M., Marchenko, N. D., Jimenez, G. S., Moll, U. M., Hope, T. J. and Wahl, G. M. (1999). A leucine-rich nuclear export signal in the p53 tetramerization domain: regulation of subcellular localization and p53 activity by NES masking. EMBO J. 18: 1660–1672.PubMedCrossRefGoogle Scholar
  61. Tanner, S. and Barberis, A. (2004). CP-31398, a putative p53-stabilizing molecule tested in mammalian cells and in yeast for its effects on p53 transcriptional activity. J. Negat. Results Biomed. 3: 5.PubMedCrossRefGoogle Scholar
  62. Vogelstein, B., Lane, D. and Levine, A. J. (2000). Surfing the p53 network. Nature 408: 307–310.PubMedCrossRefGoogle Scholar
  63. Vousden, K. H. and Lu, X. (2002). Live or let die: the cell’s response to p53. Nat. Rev. Cancer 2: 594–604.PubMedCrossRefGoogle Scholar
  64. Walker, K. K. and Levine, A. J. (1996). Identification of a novel p53 functional domain that is necessary for efficient growth suppression. Proc. Natl. Acad. Sci. USA 93: 15335–15340.PubMedCrossRefGoogle Scholar
  65. Wang, W., Takimoto, R., Rastinejad, F. and El-Deiry, W. S. (2003). Stabilization of p53 by CP-31398 inhibits ubiquitination without altering phosphorylation at serine 15 or 20 or MDM2 binding. Mol. Cell. Biol. 23: 2171–81.PubMedCrossRefGoogle Scholar
  66. Weinberg, R. L., Freund, S. M., Veprintsev, D. B., Bycroft, M. and Fersht, A. R. (2004). Regulation of DNA binding of p53 by its C-terminal domain. J. Mol. Biol. 342: 801–11.PubMedCrossRefGoogle Scholar
  67. Wong, K. B., DeDecker, B. S., Freund, S. M., Proctor, M. R., Bycroft, M. and Fersht, A. R. (1999). Hot-spot mutants of p53 core domain evince characteristic local structural changes. Proc. Natl. Acad. Sci. USA 96: 8438–8442.PubMedCrossRefGoogle Scholar
  68. Zhang, W., Guo, X. Y., Hu, G. Y., Liu, W. B., Shay, J. W. and Deisseroth, A. B. (1994). A temperature-sensitive mutant of human p53. EMBO J. 13: 2535–2544.PubMedGoogle Scholar
  69. Zhao, K., Chai, X., Johnston, K., Clements, A. and Marmorstein, R. (2001). Crystal structure of the mouse p53 core DNA-binding domain at 2.7 A resolution. J. Biol. Chem. 276: 12120–12127.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Andreas C. Joerger
    • 1
  • Assaf Friedler
    • 1
  • Alan R. Fersht
    • 1
  1. 1.MRC Centre for Protein EngineeringCambridgeUK

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