Negative regulation of p53 family protein p73 by transcription factor E2F-1

• Project/Area Number: 15590261
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Chiba Cancer Center Research Institute
• Principal Investigator: OZAKI Toshinori Chiba Cancer Center Research Institute, Division of Biochemistry, Senior Researcher, 生化学研究部, 上席研究員 (40260252)
• Co-Investigator(Kenkyū-buntansha): NAKAGAWARA Akira Chiba Cancer Center Research Institute, Director, 研究局, 局長 (50117181)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000); Fiscal Year 2003: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: p53 / p73 / E3 ligase / ubiquitin / proteasome / RanBPM / UFD2a / apoptosis / E2F1 / NEDL2 / ubiquitination

Research Abstract

We have found for the first time that E2F-1 has an ability to promote the ubiquitin-dependent proteolytic degradation of p73 in COS7 and SAOS-2 cells. Similar results were also obtained in H1299, U2OS, H4 and A549 cells. Deletion analysis revealed that the transactivation function of E2F-1 is required for the degradation of p73. We also demonstrated that a novel HECT-type E3 ubiquitin protein ligase termed NEDL2 interacts with the PY motif of p73, and thereby inducing its ubiquitination. Unexpectedly, NEDL2-mediated ubiquitination of p73 resulted in an incre ase in its half-life, and enhanced its transcriptional activity. These observations strongly suggest that there exists a non-proteolytic regulatory function of the ubiquitination. By using a yeast-based two-hybrid screening, we have identified RanBPM as a binding partner of p73. RanBPM bound to the extreme COOH-terminal region of p73, and increased its stability by inhibiting the ubiquitination levels of p73. It is likely that the COOH-terminal Lys residues could be target(s) for ubiquitination. In addition, we have found that U-box-type E3/E4 ubiquitin protein ligase termed UFD2a induces the proteasome-dependent proteolytic degradation of p73 through the physical interaction with p73. Of note, UFD2a-mediated ubiquitination of p73 was not detected under our experimental conditions. Thus, our present results indicate that p73 is regulated not only by the ubiquitination-dependent degradation pathway but also by the functional interaction with UFD2a in a ubiquitination-independent manner.

Research Products

• [Journal Article] Identification of protein kinase A catalytic subunit β(PKA-Cβ) as a novel binding partner of p73 and regulation of p73 function. (2005)
  • Author(s): Hanamoto, T., Ozaki, T., et al.
  • Journal Title: J.Biol.Chem. 280 Pages: 16665-16675
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Protein stability and function of p73 are modulated by a physical interaction with RanBPM in mammalian cultured cells. (2005)
  • Author(s): Kramer, S., Ozaki, T., et al.
  • Journal Title: Oncogene 24 Pages: 938-944
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Identification of protein kinase A catalytic subunit β (PKA-Cβ) as a novel binding partner of p73 and regulation of p73 function. (2005)
  • Author(s): Hanamoto, T., Ozaki, T., Furuya, K., Hosoda, M., Hayashi, S., Nakanishi, M., Yamamoto, H., Kikuchi, H., Todo, S., Nakagawara, A.
  • Journal Title: J.Biol.Chem. 280 Pages: 16665-16675
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Protein stability and function of p73 are modulated by a physical interaction with RanBPM in mammalian cultured cells. (2005)
  • Author(s): Kramer, S., Ozaki, T., Miyazaki, K., Kato, C., Hanamoto, T., Nakagawara, A.
  • Journal Title: Oncogene 24 Pages: 938-944
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] LMO3 interacts with neuronal transcription factor, HEN2, and acts as an oncogene in neuroblastoma. (2005)
  • Author(s): Aoyama, M., Ozaki, T., et al.
  • Journal Title: Cancer Res. (in press)
• [Journal Article] Identification of protein kinase A catalytic subunitβ(PKA-Cβ) as a novel binding partner of p73 and regulation of p73 function. (2005)
  • Author(s): Hanamoto, T., Ozaki, T., et al.
  • Journal Title: J.Biol.Chem. (in press)
• [Journal Article] Topors, a p53 topoisomerase I-binding RING finger protein, is a co-activator of p53 in growth suppression induced by DNA damage. (2005)
  • Author(s): Lin, L., Ozaki, T., et al.
  • Journal Title: Oncogene (in press)
• [Journal Article] Polo-like kinase 1 (Plk1) inhibits p53 function by physical interaction and phosphorylation. (2004)
  • Author(s): Ando, K., Ozaki, T., et al.
  • Journal Title: J.Biol.Chem. 279 Pages: 25549-25561
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Polo-like kinase 1(Plk1) inhibits p53 function by physical interaction and phosphorylation. (2004)
  • Author(s): Ando, K., Ozaki, T., Yamamoto, H., Furuya, K., Hosoda, M., Hayashi, S., Fukuzawa, M., Nakagawara, A.
  • Journal Title: J.Biol.Chem. 279 Pages: 25549-25561
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Decreased expression of the candidate tumor suppressor gene ING1 is associated with poor prognosis in advanced neuroblastomas. (2004)
  • Author(s): Takahashi, M., Ozaki, T., et al.
  • Journal Title: Oncogene 12 Pages: 811-816
• [Journal Article] Polo-like Kinase 1(Plk1) inhibits p53 function by physical interaction and phosphorylation. (2004)
  • Author(s): Ando, K., Ozaki, T., et al.
  • Journal Title: J.Biol.Chem. 279 Pages: 25519-25561
• [Journal Article] A novel HECT-type E3 ubiquitin ligase, NEDL2, stabilizes p73 and enhances its transcriptional activity. (2003)
  • Author(s): Miyazaki, K., Ozaki, T., et al.
  • Journal Title: Biochem.Biophys.Res.Commun. 308 Pages: 106-113
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Negative autoregulation of p73 and p53 by DeltaNp73 in regulating differentiation and survival of human neuroblastoma cells. (2003)
  • Author(s): Nakagawa, T., Ozaki, T., et al.
  • Journal Title: Cancer Lett. 197 Pages: 105-109
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Function of p73, not of p53, is inhibited by the physical interaction with RACK1 and its inhibitory effect is counteracted by pRB. (2003)
  • Author(s): Ozaki, T., Watanabe, K., et al.
  • Journal Title: Oncogene 22 Pages: 3231-3242
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A novel HECT-type E3 ubiquitin ligase, NEDL2,stabilizes p73 and enhances its transcriptional activity. (2003)
  • Author(s): Miyazaki, K., Ozaki, T., Kato, C., Hanamoto, T., Fujita, T., Irino, S., Watanabe, K., Nakagawa, T., Nakagawara, A.
  • Journal Title: Biochem.Biophys.Res.Commun. 308 Pages: 106-113
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Negative autoregulation of p73 and p53 by DeltaNp73 in regulating differentiation and survival of human neuroblastoma cells. (2003)
  • Author(s): Nakagawa, T., Takahashi, M., Ozaki, T., Watanabe, K., Hayashi, S., Hosoda, M., Todo, S., Nakagawara, A.
  • Journal Title: Cancer Lett. 197 Pages: 105-109
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Function of p73,not of p53,is inhibited by the physical interaction with RACK1 and its inhibitory effect is counteracted by pRB. (2003)
  • Author(s): Ozaki, T., Watanabe, K., Nakagawa, T., Miyazaki, K., Takahashi, M., Nakagawara, A.
  • Journal Title: Oncogene 22 Pages: 3231-3242
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Miyazaki, K., Fujita, T., Ozaki, T., et al.: "NEDL1, a novel E3 Ubiquitin ligase for dishevelled1, targets mutant super oxide dismutase 1."J.Biol.Chem.. 279. 11327-11335 (2004)
• [Publications] Miyazaki, K., Ozaki, T., et al.: "A novel HECT-type E3 ubiquitin ligase, NEDL2, stabilizes p73 and enhances its transcriptional activity."Biochem.Biophys.Res.Commun.. 308. 106-113 (2003)
• [Publications] Nakagawa, T., Takahashi, M., Ozaki, T., et al.: "Negative auto regulation of p73 and p53 by DeltaNp73 in regulating differentiation and survival of human neuroblastoma cells."Cancer Lett.. 197. 105-109 (2003)
• [Publications] Okamoto, Y., Ozaki, T., et al.: "UbcH10 is the cancer-related E2 ubiquitin-conjugating enzyme."Cancer Res.. 63. 4167-4173 (2003)
• [Publications] Nakamura, Y., Ozaki, T., et al.: "Accumulation of p27^<KIP1> is associated with BMP2-induced growth arrest and neuronal differentiation of human neuroblastoma-derived cell lines."Biochem.Biophys.Res.Commun.. 307. 206-213 (2003)
• [Publications] Ozaki, T., Watanabe, K., et al.: "Function of p73, not of p53, is inhibited by the physical interaction with RACK1 and its inhibitory effect is counteracted by pRB."Oncogene. 22. 3231-8242 (2003)