Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

A novel HECT-type E3 ubiquitin protein ligase NEDL1 enhances the p53-mediated apoptotic cell death in its catalytic activity-independent manner

Oncogene volume 27, pages 3700–3709 (2008)Cite this article

Abstract

NEDL1 (NEDD4-like ubiquitin protein ligase-1) is a newly identified HECT-type E3 ubiquitin protein ligase highly expressed in favorable neuroblastomas as compared with unfavorable ones. In this study, we found that NEDL1 cooperates with p53 to induce apoptosis. During cisplatin (CDDP)-mediated apoptosis in neuroblastoma SH-SY5Y cells, p53 was induced to accumulate in association with an increase in expression levels of NEDL1. Enforced expression of NEDL1 resulted in a decrease in number of G418-resistant colonies in SH-SY5Y and U2OS cells bearing wild-type p53, whereas NEDL1 had undetectable effect on p53-deficient H1299 and SAOS-2 cells. Similarly, enforced expression of NEDL1 increased number of U2OS cells with sub-G1 DNA content. Co-immunoprecipitation and in vitro binding assays revealed that NEDL1 binds to the COOH-terminal region of p53. Luciferase reporter assay showed that NEDL1 has an ability to enhance the transcriptional activity of p53. Small interfering RNA-mediated knockdown of the endogenous NEDL1 conferred the resistance of U2OS cells to adriamycin. It is noteworthy that NEDL1 enhanced pro-apoptotic activity of p53 in its catalytic activity-independent manner. Taken together, our present findings suggest that functional interaction of NEDL1 with p53 might contribute to the induction of apoptosis in cancerous cells bearing wild-type p53.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  • Adhikary S, Marinoni F, Hock A, Hulleman E, Popov N, Beier R et al. (2005). The ubiquitin ligase HectH9 regulates transcriptional activation by Myc and is essential for tumor cell proliferation. Cell 123: 409–421.

    Article  CAS  Google Scholar 

  • Bijur GN, De Sarno P, Jope RS . (2000). Glycogen synthase kinase-3beta facilitates staurosporine- and heat shock-induced apoptosis. J Biol Chem 275: 7583–7590.

    Article  CAS  Google Scholar 

  • Cluskey S, Ramsden DB . (2001). Mechanisms of neurodegeneration in amyotrophic lateral sclerosis. Mol Pathol 54: 386–392.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Cross DA, Culbert AA, Chalmers KA, Facci L, Skaper SD, Reith AD . (2001). Selective small-molecule inhibitors of glycogen synthase kinase-3 activity protect primary neurones from death. J Neurochem 77: 94–102.

    Article  CAS  Google Scholar 

  • Di Lello P, Jenkins LM, Jones TN, Nguyen BD, Hara T, Yamaguchi H et al. (2006). Structure of the Tfb1/p53 complex: Insights into the interaction between the p62/Tfb1 subunit of TFIIH and the activation domain of p53. Mol Cell 22: 731–740.

    Article  CAS  Google Scholar 

  • Donehower LA, Harvey M, Slagle BL, McArthur MJ, Montgomery Jr CA, Butel JS et al. (1992). Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours. Nature 356: 215–221.

    Article  CAS  Google Scholar 

  • Ekegren T, Grundstrom E, Lindholm D, Aquilonius SM . (1999). Upregulation of Bax protein and increased DNA degradation in ALS spinal cord motor neurons. Acta Neurol Scand 100: 317–321.

    Article  CAS  Google Scholar 

  • Gonzalez de Aguilar JL, Gordon JW, Rene F, de Tapia M, Lutz-Bucher B, Gaiddon C et al. (2000). Alteration of the Bcl-x/Bax ratio in a transgenic mouse model of amyotrophic lateral sclerosis: evidence for the implication of the p53 signaling pathway. Neurobiol Dis 7: 406–415.

    Article  CAS  Google Scholar 

  • Hetman M, Cavanaugh JE, Kimelman D, Xia Z . (2000). Role of glycogen synthase kinase-3beta in neuronal apoptosis induced by trophic withdrawal. J Neurosci 20: 2567–2574.

    Article  CAS  Google Scholar 

  • Hino S, Michiue T, Asashima M, Kikuchi A . (2003). Casein kinase I epsilon enhances the binding of Dvl-1 to Frat-1 and is essential for Wnt-3a-induced accumulation of beta-catenin. J Biol Chem 278: 14066–14073.

    Article  CAS  Google Scholar 

  • Hollstein M, Sidransky D, Vogelstein B, Harris CC . (1991). p53 mutations in human cancers. Science 253: 49–53.

    Article  CAS  Google Scholar 

  • Hupp TR, Lane DP . (1994). Regulation of the cryptic sequence-specific DNA-binding function of p53 by protein kinases. Cold Spring Harb Symp Quant Biol 59: 195–206.

    Article  CAS  Google Scholar 

  • Kiryo-Seo S, Hirayama T, Kato R, Kiyama H . (2005). Noxa is a critical mediator of p53-dependent motor neuron death after nerve injury in adult mouse. J Neurosci 25: 1442–1447.

    Article  Google Scholar 

  • Kishida M, Hino S, Michiue T, Yamamoto H, Kishida S, Fukui A et al. (2001). Synergistic activation of the Wnt signaling pathway by Dvl and casein kinase Iepsilon. J Biol Chem 276: 33147–33155.

    Article  CAS  Google Scholar 

  • Kitanaka C, Kato K, Ijiri R, Sakurada K, Tomiyama A, Noguchi K et al. (2002). Increased Ras expression and caspase-independent neuroblastoma cell death: possible mechanism of spontaneous neuroblastoma regression. J Natl Cancer Inst 94: 358–368.

    Article  CAS  Google Scholar 

  • Lee E, Salic A, Kirschner MW . (2001). Physiological regulation of [beta]- catenin stability by Tcf3 and CK1epsilon. J Cell Biol 154: 983–993.

    Article  CAS  Google Scholar 

  • Levine AJ, Chang A, Dittmer D, Notterman DA, Silver A, Thorn K et al. (1994). The p53 tumor suppressor gene. J Lab Clin Med 123: 817–823.

    CAS  PubMed  Google Scholar 

  • Martin LJ . (2000). p53 is abnormally elevated and active in the CNS of patients with amyotrophic lateral sclerosis. Neurobiol Dis 7: 613–622.

    Article  CAS  Google Scholar 

  • Martin LJ, Liu Z . (2002). Injury-induced spinal motor neuron apoptosis is preceded by DNA single-strand breaks and is p53- and bax-dependent. J Neurobiol 5: 181–197.

    Article  Google Scholar 

  • Miyazaki K, Fujita T, Ozaki T, Kato C, Kurose Y, Sakamoto M et al. (2004). NEDL1, a novel ubiquitin-protein isopeptide ligase for dishevelled-1, targets mutant superoxide dismutase-1. J Biol Chem 279: 11327–11335.

    Article  CAS  Google Scholar 

  • Miyazaki K, Ozaki T, Kato C, Hanamoto T, Fujita T, Irino S et al. (2003). A novel HECT-type E3 ubiquitin ligase, NEDL2, stabilizes p73 and enhances its transcriptional activity. Biochem Biophys Res Commun 308: 106–113.

    Article  CAS  Google Scholar 

  • Moll UM, LaQuaglia M, Benard J, Riou G . (1995). Wild-type p53 protein undergoes cytoplasmic sequestration in undifferentiated neuroblastomas but not in differentiated tumors. Proc Natl Acad Sci USA 92: 4407–44011.

    Article  CAS  Google Scholar 

  • Nakagawara A, Ohira M . (2004). Comprehensive genomics linking between neural development and cancer: neuroblastoma as a model. Cancer Lett 204: 213–224.

    Article  CAS  Google Scholar 

  • Pietenpol JA, Tokino T, Thiagalingam S, el-Deiry WS, Kinzler KW, Vogelstein B . (1994). Sequence-specific transcriptional activation is essential for growth suppression by p53. Proc Natl Acad Sci USA 91: 1998–2002.

    Article  CAS  Google Scholar 

  • Roos WP, Kaina B . (2006). DNA damage-induced cell death by apoptosis. Trends Mol Med 12: 440–450.

    Article  CAS  Google Scholar 

  • Shaw P, Freeman J, Bovey R, Iggo R . (1996). Regulation of specific DNA binding by p53: evidence for a role for O-glycosylation and charged residues at the carboxy-terminus. Oncogene 12: 921–930.

    CAS  PubMed  Google Scholar 

  • Thomas MC, Chiang CM . (2005). E6 oncoprotein represses p53-dependent gene activation via inhibition of protein acetylation independently of inducing p53 degradation. Mol Cell 17: 251–264.

    Article  CAS  Google Scholar 

  • Vousden KH, Lu X . (2002). Live or let die: the cell's response to p53. Nat Rev Cancer 2: 594–604.

    Article  CAS  Google Scholar 

  • Watcharasit P, Bijur GN, Zmijewski JW, Song L, Zmijewska A, Chen X et al. (2002). Direct, activating interaction between glycogen synthase kinase-3beta and p53 after DNA damage. Proc Natl Acad Sci USA 99: 7951–7955.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to Dr T Kamijo (Division of Biochemistry, Chiba Cancer Center Research Institute) for his helpful discussion. This work was supported in part by a grant-in-aid from the Ministry of Health, Labour and Welfare for Third Term Comprehensive Control Research for Cancer, a grant-in-aid for Scientific Research on Priority Areas from the Ministry of Education, Culture, Sports, Science and Technology, Japan, a grant-in-aid for Scientific Research from Japan Society for the Promotion of Science and Uehara Memorial Foundation.

Author information

Author notes

  1. Y Li and T Ozaki: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Biochemistry, Chiba Cancer Center Research Institute, Nitona, Chuou-Ku, Chiba, Japan

    Y Li, T Ozaki, H Kikuchi, H Yamamoto, M Ohira & A Nakagawara

  2. Production Technology Development Center, the Furukawa Electric Co., Ltd., 6 Yawata-Kaigandori, Ichihara, Japan

    Y Li

Authors
  1. Y Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T Ozaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. H Kikuchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. H Yamamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M Ohira
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. A Nakagawara
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to A Nakagawara.

Additional information

Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, Y., Ozaki, T., Kikuchi, H. et al. A novel HECT-type E3 ubiquitin protein ligase NEDL1 enhances the p53-mediated apoptotic cell death in its catalytic activity-independent manner. Oncogene 27, 3700–3709 (2008). https://doi.org/10.1038/sj.onc.1211032

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1211032

Keywords

This article is cited by

Search

Advanced search

Quick links