In vivo effects of abolishing the Single canonical sumoylation site in the C-terminal region of Drosophila p53


Using yeast two-hybrid screens we determined that Drosophila (Dm)p53 interacts with proteins involved in sumoylation (UBA2, UBC9 and PIAS) through different regions of its C-terminal domain. A K302R point mutation within a single canonical sumoylation site of Dmp53 did not abolish the observed interactions. These observations prompted us to analyze whether Dmp53 sumoylation at this site has any functional role in vivo. Genetic assays showed that deleting one copy of genes involved in sumoylation (lwr, Su(var)2–10 or smt3 heterozygosity) enhanced slightly the mutator phenotype of Dmp53. We compared the in vivo effects of wild type and K302R Dmp53 overproduced from transgenes and determined that similar levels of expression of the mutant and wild type proteins resulted in similar phenotype, and the two proteins showed similar cellular localization. The half life and the trans-activator activity of K302R mutant and wild type Dmp53 were also comparable. Lastly, by analyzing wild type and K302R Dmp53 expressed at different levels in animals and in S2 cells we detected no differences between the mobility of the mutant and wild-type protein. From these data we conclude that under normal developmental conditions the loss of SUMO modification at K302 does not affect Dmp53 function significantly.



Basic Local Alignment Search Tool


fluorescence activated cell sorting


gluthation S-transferase


multiple wing hair


Trp-, Ura-, Leu-, Lys-, His-


yeast two-hybrid


  1. 1.

    Altschul, S. F., Madden, T. L., Schaffer, A. A., Zhang, J., Zhang, Z., Miller, W., Lipman, D. J. (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25, 3389–3402.

    CAS  PubMed  PubMed Central  Google Scholar 

  2. 2.

    Bereczki, O., Ujfaludi, Z., Pardi, N., Nagy, Z., Tora, L., Boros, I. M., Balint, E. (2008) TATA binding protein associated factor 3 (TAF 3) interacts with p53 and inhibits its function. BMC Mol. Biol. 9, 57.

    Article  Google Scholar 

  3. 3.

    Bischof, J., Maeda, R. K., Hediger, M., Karch, F., Basler, K. (2007) An optimized transgenesis system for Drosophila using germ-line-specific phiC31 integrases. Proc. Natl. Acad. Sci. (USA) 104, 3312–3317.

    CAS  Article  Google Scholar 

  4. 4.

    Bodai, L., Pardi, N., Ujfaludi, Z., Bereczki, O., Komonyi, O., Balint, E., Boros, I. M. (2007) Daxxlike protein of Drosophila interacts with Dmp53 and affects longevity and Ark mRNA level. J. Biol. Chem. 282, 36386–36393.

    CAS  Article  Google Scholar 

  5. 5.

    Breeden, L. Nasmyth, K. (1985) Regulation of the yeast HO gene. Cold Spring Harb. Symp. Quant. Biol. 50, 643–650.

    CAS  Article  Google Scholar 

  6. 6.

    Brodsky, M. H., Nordstrom, W., Tsang, G., Kwan, E., Rubin, G. M., Abrams, J. M. (2000) Drosophila p53 binds a damage response element at the reaper locus. Cell 101, 103–113.

    CAS  Article  Google Scholar 

  7. 7.

    Formstecher, E., Aresta, S., Collura, V., Hamburger, A., Meil, A., Trehin, A., Reverdy, C., Betin, V., Maire, S., Brun, C., Jacq, B., Arpin, M., Bellaiche, Y., Bellusci, S., Benaroch, P., Bornens, M., Chanet, R., Chavrier, P., Delattre, O., Doye, V., Fehon, R., Faye, G., Galli, T., Girault, J. A., Goud, B., de Gunzburg, J., Johannes, L., Junier, M. P., Mirouse, V., Mukherjee, A., Papadopoulo, D., Perez, F., Plessis, A., Rosse, C., Saule, S., Stoppa-Lyonnet, D., Vincent, A., White, M., Legrain, P., Wojcik, J., Camonis, J., Daviet, L. (2005) Protein interaction mapping: a Drosophila case study. Genome Res. 15, 376–384.

    CAS  Article  Google Scholar 

  8. 8.

    Gostissa, M., Hengstermann, A., Fogal, V., Sandy, P., Schwarz, S. E., Scheffner, M., Del Sal, G. (1999) Activation of p53 by conjugation to the ubiquitin-like protein SUMO-1. Embo J. 18, 6462–6471.

    CAS  Article  Google Scholar 

  9. 9.

    Jin, S., Martinek, S., Joo, W. S., Wortman, J. R., Mirkovic, N., Sali, A., Yandell, M. D., Pavletich, N. P., Young, M. W., Levine, A. J. (2000) Identification and characterization of a p53 homologue in Drosophila melanogaster. Proc. Natl. Acad. Sci. (USA) 97, 7301–7306.

    CAS  Article  Google Scholar 

  10. 10.

    Johnson, E. S. (2004) Protein modification by SUMO. Annu. Rev. Biochem. 73, 355–382.

    CAS  Article  Google Scholar 

  11. 11.

    Kaddurah-Daouk, R., Greene, J. M., Baldwin, A. S., Jr., Kingston, R. E. (1987) Activation and repression of mammalian gene expression by the c-myc protein. Genes Dev. 1, 347–357.

    CAS  Article  Google Scholar 

  12. 12.

    Kwek, S. S., Derry, J., Tyner, A. L., Shen, Z., Gudkov, A. V. (2001) Functional analysis and intracellular localization of p53 modified by SUMO-1. Oncogene 20, 2587–2599.

    CAS  Article  Google Scholar 

  13. 13.

    Mauri, F., McNamee, L. M., Lunardi, A., Chiacchiera, F., Del Sal, G., Brodsky, M. H., Collavin, L. (2008) Modification of Drosophila p53 by SUMO modulates its transactivation and pro-apoptotic functions. J. Biol. Chem. 283, 20848–20856.

    CAS  Article  Google Scholar 

  14. 14.

    Melchior, F. Hengst, L. (2002) SUMO-1 and p53. Cell Cycle 1, 245–249.

    CAS  Article  Google Scholar 

  15. 15.

    Minty, A., Dumont, X., Kaghad, M., Caput, D. (2000) Covalent modification of p73alpha by SUMO- 1. Two-hybrid screening with p73 identifies novel SUMO-1-interacting proteins and a SUMO-1 interaction motif. J. Biol. Chem. 275, 36316–36323.

    CAS  Article  Google Scholar 

  16. 16.

    Muller, S., Berger, M., Lehembre, F., Seeler, J. S., Haupt, Y., Dejean, A. (2000) c-Jun and p53 activity is modulated by SUMO-1 modification. J. Biol. Chem. 275, 13321–13329.

    CAS  Article  Google Scholar 

  17. 17.

    Muller, S., Ledl, A., Schmidt, D. (2004) SUMO: a regulator of gene expression and genome integrity. Oncogene 23, 1998–2008.

    Article  Google Scholar 

  18. 18.

    Ollmann, M., Young, L. M., Di Como, C. J., Karim, F., Belvin, M., Robertson, S., Whittaker, K., Demsky, M., Fisher, W. W., Buchman, A., Duyk, G., Friedman, L., Prives, C., Kopczynski, C. (2000) Drosophila p53 is a structural and functional homolog of the tumor suppressor p53. Cell 101, 91–101.

    CAS  Article  Google Scholar 

  19. 19.

    Pankotai, T., Komonyi, O., Bodai, L., Ujfaludi, Z., Muratoglu, S., Ciurciu, A., Tora, L., Szabad, J., Boros, I. (2005) The homologous Drosophila transcriptional adaptors ADA2a and ADA2b are both required for normal development but have different functions. Mol. Cell. Biol. 25, 8215–8227.

    CAS  Article  Google Scholar 

  20. 20.

    Rodriguez, M. S., Desterro, J. M., Lain, S., Midgley, C. A., Lane, D. P., Hay, R. T. (1999) SUMO-1 modification activates the transcriptional response of p53. Embo J. 18, 6455–6461.

    CAS  Article  Google Scholar 

  21. 21.

    Rong, Y. S., Titen, S. W., Xie, H. B., Golic, M. M., Bastiani, M., Bandyopadhyay, P., Olivera, B. M., Brodsky, M., Rubin, G. M., Golic, K. G. (2002) Targeted mutagenesis by homologous recombination in D. melanogaster. Genes Dev. 16, 1568–1581.

    CAS  Article  Google Scholar 

  22. 22.

    Sogame, N., Kim, M., Abrams, J. M. (2003) Drosophila p53 preserves genomic stability by regulating cell death. Proc. Natl. Acad. Sci. (USA) 100, 4696–4701.

    CAS  Article  Google Scholar 

  23. 23.

    Stanyon, C. A., Liu, G., Mangiola, B. A., Patel, N., Giot, L., Kuang, B., Zhang, H., Zhong, J., Finley, R. L., Jr. (2004) A Drosophila protein-interaction map centered on cell-cycle regulators. Genome Biol. 5, R96.

    Article  Google Scholar 

  24. 24.

    Talamillo, A., Sanchez, J., Barrio, R. (2008) Functional analysis of the SUMOylation pathway in Drosophila. Biochem. Soc. Trans. 36, 868–873.

    CAS  Article  Google Scholar 

Download references

Author information



Corresponding author

Correspondence to I. M. Boros.

Rights and permissions

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Reprints and Permissions

About this article

Cite this article

Pardi, N., Vámos, E., Ujfaludi, Z. et al. In vivo effects of abolishing the Single canonical sumoylation site in the C-terminal region of Drosophila p53. BIOLOGIA FUTURA 62, 397–412 (2011).

Download citation


  • p53
  • SUMO
  • apoptosis
  • Drosophila
  • yeast two hybrid