The ubiquitin–protein ligase Itch regulates p73 stability

Mario Rossi, Vincenzo De Laurenzi, Eliana Munarriz, Douglas R Green, Yun-Cai Liu, Karen H Vousden, Gianni Cesareni and Gerry Melino


The EMBO Journal (2005) 24, 836–848


Читать статью


p73, a member of the p53 family of transcription factors, is upregulated in response to DNA damage, inducing cell cycle arrest and apoptosis. Besides indications that this p73 response is post-transcriptional, little is known about the underlying molecular mechanisms of p73 protein degradation. Ubiquitination and proteasomal-dependent degradation of p53 are regulated by its transcriptional target MDM2. However, unlike p53, p73 binds to, but is not degraded by, MDM2. Here we describe the binding of p73 to Itch, a Hect ubiquitin–protein ligase. Itch selectively binds and ubiquitinates p73 but not p53; this results in the rapid proteasome-dependent degradation of p73. Upon DNA damage Itch itself is downregulated, allowing p73 protein levels to rise and thus interfere with p73 function. In conclusion, we have identified a key mechanism in the control of p73 protein levels both in normal as well as in stress conditions.


Agami R, Blandino G, Oren M, Shaul Y (1999) Interaction of c-Abl and p73alpha and their collaboration to induce apoptosis. Nature 399: 809–813

Balint E, Bates S, Vousden KH (1999) Mdm2 binds p73 alpha without targeting degradation. Oncogene 18: 3923–3929

Bernassola F, Salomoni P, Oberst A, Di Como CJ, Pagano M, Melino G, Pandolfi PP (2004) Ubiquitin-dependent degradation of p73 is inhibited by PML. J Exp Med 199: 1545–1557

Casciano I, Mazzocco K, Boni L, Pagnan G, Banelli B, Allemanni G, Ponzoni M, Tonini GP, Romani M (2002) Expression of DeltaNp73 is a molecular marker for adverse outcome in neuroblastoma patients. Cell Death Differ 9: 246–251

Castagnoli L, Zucconi A, Quondam M, Rossi M, Vaccaro P, Panni S, Paoluzi S, Santonico E, Dente L, Cesareni G (2001) Alternative bacteriophage display systems. Comb Chem High Throughput Screen 4: 121–133

Catani MV, Costanzo A, Savini I, Levrero M, de Laurenzi V, Wang JY, Melino G, Avigliano L (2002) Ascorbate up-regulates MLH1 (Mut L homologue-1) and p73: implications for the cellular response to DNA damage. Biochem J 364: 441–447

Cesareni G, Castagnoli L, Cestra G (1999) Phage displayed peptide libraries. Comb Chem High Throughput Screen 2: 1–17

De Laurenzi V, Catani MV, Terrinoni A, Corazzari M, Melino G, Costanzo A, Levrero M, Knight RA (1999) Additional complexity in p73: induction by mitogens in lymphoid cells and identification of two new splicing variants epsilon and zeta. Cell Death Differ 6:

De Laurenzi V, Costanzo A, Barcaroli D, Terrinoni A, Falco M, Annicchiarico-Petruzzelli M, Levrero M, Melino G (1998) Two new p73 splice variants, gamma and delta, with different transcriptional activity. J Exp Med 188: 1763–1768

De Laurenzi V, Raschella G, Barcaroli D, Annicchiarico-Petruzzelli M, Ranalli M, Catani MV, Tanno B, Costanzo A, Levrero M, Melino G (2000) Induction of neuronal differentiation by p73, in a neuroblastoma cell line. Journal Biological Chemistry 275: 15226–15231

Dobbelstein M, Wienzek S, Konig C, Roth J (1999) Inactivation of the p53-homologue p73 by the mdm2-oncoprotein. Oncogene 18: 2101–2106

Fang D, Elly C, Gao B, Fang N, Altman Y, Joazeiro C, Hunter T, Copeland N, Jenkins N, Liu YC (2002) Dysregulation of T lymphocyte function in itchy mice: a role for Itch in TH2 differentiation. Nat Immunol 3: 281–287

Gong JC, Costanzo A, Yang HQ, Melino G, Kaelin JrWG, Levrero M, Wang JY (1999) The tyrosine kinase c-Abl regulates p73 in apoptotic response to cisplatin-induced DNA damage. Nature 399: 806–809

Gottifredi V, Pelicci G, Munarriz E, Maione R, Pelicci PG, Amati P (1999) Polyomavirus large T antigen induces alterations in cytoplasmic signalling pathways involving Shc activation. J Virol 73: 1427–1437

Grob TJ, Novak U, Maisse C, Barcaroli D, Luthi AU, Pirnia F, Hugli B, Graber HU, De Laurenzi V, Fey MF, Melino G, Tobler A (2001)

Human delta Np73 regulates a dominant negative feedback loop for TAp73 and p53. Cell Death Differ 8: 1213–1223

Hamilton MH, Tcherepanova I, Huibregtse JM, McDonnell DP (2001) Nuclear import/export of hRPF1/Nedd4 regulates the ubiquitin-dependent degradation of its nuclear substrates. J Biol Chem 276: 26324–26331

Harvey KF, Kumar S (1999) Nedd4-like proteins: an emerging family of ubiquitin-protein ligases implicated in diverse cellular functions. Trends Cell Biol 9: 166–169

Hicke L (2001) Protein regulation by monoubiquitin. Nat Rev Mol Cell Biol 2: 195–201

Ikawa S, Nakagawara A, Ikawa Y (1999) p53 family genes: structural comparison, expression and mutation. Cell Death Differ 6: 1154–1161

Kaghad M, Bonnet H, Yang A, Creancier L, Biscan JC, Valent A, Minty A, Chalon P, Lelias JM, Dumont X, Ferrara P, McKeon F, Caput D (1997) Monoallelically expressed gene related to p53 at 1p36, a region frequently deleted in neuroblastoma and other human cancers. Cell 90: 809–819

Kloetzel PM (2001) Antigen processing by the proteasome. Nat Rev Mol Cell Biol 2: 179–187

Kumar S, Kao WH, Howley PM (1997) Physical interaction between specific E2 and Hect E3 enzymes determines functional cooperativity. J Biol Chem 272: 13548–13554

Lohrum MA, Vousden KH (1999) Regulation and activation of p53 and its family members. Cell Death Differ 6: 1162–1168

Maisse C, Munarriz E, Barcaroli D, Melino G, De Laurenzi V (2004) DNA damage induces the rapid and selective degradation of the DeltaNp73 isoform, allowing apoptosis to occur. Cell Death Differ 11: 685–687

Melino G, Bernassola F, Ranalli M, Yee K, Zong WX, Corazzari M, Knight RA, Green DR, Thompson C, Vousden KH (2004) p73 Induces apoptosis via PUMA transactivation and Bax mitochondrial translocation. J Biol Chem 279: 8076–8083

Melino G, De Laurenzi V, Vousden KH (2002) p73: Friend or foe in tumorigenesis. Nat Rev Cancer 2: 605–615

Melino G, Lu X, Gasco M, Crook T, Knight RA (2003) Functional regulation of p73 and p63: development and cancer. Trends Biochem Sci 28: 663–670

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

Nakano K, Balint E, Ashcroft M, Vousden KH (2000) A ribonucleotide reductase gene is a transcriptional target of p53 and p73. Oncogene 19: 4283–4289

Ongkeko WM, Wang XQ, Siu WY, Lau AW, Yamashita K, Harris AL, Cox LS, Poon RY (1999) MDM2 and MDMX bind and stabilize the p53-related protein p73. Curr Biol 9: 829–832

Perry WL, Hustad CM, Swing DA, O’Sullivan TN, Jenkins NA, Copeland NG (1998) The itchy locus encodes a novel ubiquitin pro-tein ligase that is disrupted in a18H mice. Nat Genet 18: 143–146

Putzer BM, Tuve S, Tannapfel A, Stiewe T (2003) Increased DeltaNp73 expression in tumors by upregulation of the E2F1-regulated, TA-promoter-derived DeltaN0-p73 transcript. Cell Death Differ 10: 612–614

Qiu L, Joazeiro C, Fang N, Wang HY, Elly C, Altman Y, Fang D, Hunter T, Liu YC (2000) Recognition and ubiquitination of Notch by Itch, a hect-type E3 ubiquitin ligase. J Biol Chem 275: 35734–35737

Romani M, Tonini GP, Banelli B, Allemanni G, Mazzocco K, Scaruffi P, Boni L, Ponzoni M, Pagnan G, Raffaghello L, Ferrini S, Croce M, Casciano I (2003) Biological and clinical role of p73 in neuroblastoma. Cancer Lett 197: 111–117

Sayan AE, Rossi M, Melino G, Knight RA (2004) p73: in silico evidence for a putative third promoter region. Biochem Biophys Res Commun 313: 765–770

Stiewe T, Putzer BM (2002) Role of p73 in malignancy: tumor suppressor or oncogene? Cell Death Differ 9: 237–245

Strano S, Munarriz E, Rossi M, Castagnoli L, Shaul Y, Sacchi A, Oren M, Sudol M, Cesareni G, Blandino G (2001) Physical interaction with Yes-associated protein enhances p73 transcriptional activity. J Biol Chem 276: 15164–15173

Sudol M (1996) Structure and function of the WW domain. Prog Biophys Mol Biol 65: 113–132

Toh WH, Siddique MM, Boominathan L, Lin KW, Sabapathy K (2004) c-Jun regulates the stability and activity of the p53 homologue, p73. Advance online publication 9 August 2004; DOI:10.1074/JBC.M407672200

Treier M, Staszewski LM, Bohmann D (1994) Ubiquitin-dependent c-Jun degradation in vivo is mediated by the delta domain. Cell 78: 787–798

Tschan MP, Grob TJ, Peters UR, De Laurenzi V, Huegli B, Kreuzer KA, Schmidt CA, Melino G, Fey MF, Tobler A, Cajot J-F (2000) Enhanced p73 expression during differentiation and complex p73 isoforms in myeloid leukemia. Biochemical Biophysical Research Communications 277: 62–65

Ueda Y, Hijikata M, Takagi S, Chib T, Shimotohno K (1999) New p73 variants with altered C-terminal structures have varied transcriptional activities. Oncogene 18: 4993–4998

Vossio S, Palescandolo E, Pediconi N, Moretti F, Balsano C, Levrero M, Costanzo A (2002) DN-p73 is activated after DNA damage in a p53-dependent manner to regulate p53-induced cell cycle arrest. Oncogene 21: 3796–3803

Weissman AM (2001) Themes and variations on ubiquitylation. Nat Rev Mol Cell Biol 2: 169–178

Winberg G, Matskova L, Chen F, Plant P, Rotin D, Gish G, Ingham R, Ernberg I, Pawson T (2000) Latent membrane protein 2A of Epstein-Barr virus bindsWWdomain E3 protein-ubiquitin ligases that ubiquitinate B-cell tyrosine kinases. Mol Cell Biol 20: 8526–8535

Yang A, Walker N, Bronson R, Kaghad M, Oosterwegel M, Bonnin J, Vagner C, Bonnet H, Dikkes P, Sharpe A, McKeon F, Caput D (2000) p73-deficient mice have neurological, pheromonal and inflammatory defects but lack spontaneous tumours. Nature 404: 99–103

Yuan ZM, Shioya H, Ishiko T, Sun X, Gu J, Huang YY, Lu H, Kharbanda S, Weichselbaum R, Kufe D (1999) p73 is regulated by tyrosine kinase c-Abl in the apoptotic response to DNA damage. Nature 399: 814–817

Zaika AI, Slade N, Erster SH, Sansome C, Joseph TW, Pearl M, Chalas E, Moll UM (2002) DeltaNp73, a dominant-negative inhibitor of wild-type p53 and TAp73, is up-regulated in human tumors. J Exp Med 196: 765–780

Zeng X, Chen L, Jost CA, Maya R, Keller D, Wang X, Kaelin Jr WG, Oren M, Chen J, Lu H (1999) MDM2 suppresses p73 function without promoting p73 degradation. Mol Cell Biol 19: 3257–3266

Zucconi A, Dente L, Santonico E, Castagnoli L, Cesareni G (2001) Selection of ligands by panning of domain libraries displayed on phage lambda reveals new potential partners of synaptojanin 1. J Mol Biol 307: 1329–1339

Прочитано 5330 раз
Оцените материал
(2 голосов)
Опубликовано в СТАТЬИ
Авторизуйтесь, чтобы получить возможность оставлять комментарии


Управление научных исследований СПбГТИ (ТУ)

Горячие новости

Приборное оснащение лаборатории

Rambler's Top100
//'+ 'Рейтинг<\/a><\/p>');})(window,navigator,document);//]]>