An assay for the autoubiquitination activity of the E3 ligaseHDM2 (Mdm2) was developed and adapted to a high-throughput format to identify inhibitors of this activity. The assay can also be used tomeasure the activity of other E3s andmay be useful in finding both inhibitors and activators of a wide range of different ubiquitin ligases.
Schwartz DC, Hochstrasser M: A superfamily of protein tags: ubiquitin, SUMO and related modifiers. Trends Biochem Sci2003;28:321-328.
2.
Glickman MH, Ciechanover A: The ubiquitin-proteasome proteolytic hway: destruction for the sake of construction. Physiol Rev2002;82:373-428.
3.
Yang Y, Yu X: Regulation of apoptosis: the ubiquitous way. FASEB J2003;17:790-799.
4.
Clarke DJ: Proteolysis and the cell cycle. Cell Cycle2002;1:233-234.
5.
Nalepa G, Wade Harper J: Therapeutic anti-cancer targets upstream of the proteasome. Cancer Treat Rev2003;29(suppl. 1):49-57.
6.
Fang S, Lorick KL, Jensen JP, Weissman AM: RING finger ubiquitin protein ligases: implications for tumorigenesis, metastasis and for molecular targets in cancer. Semin Cancer Biol2003;13:5-14.
7.
Pickart CM: Mechanisms underlying ubiquitination. Ann Rev Biochem2001;70:503-533.
8.
Hicke L: Protein regulation by monoubiquitin. Nat Rev Mol Cell Biol2001;2:195-201.
9.
Jackson PK, Eldridge AG, Freed E, et al: The lore of theRINGs: substrate recognition and catalysis by ubiquitin ligases. TrendsCell Biol2000;10:429-439.
10.
Adams J: Potential for proteasome inhibition in the treatment of cancer. Drug Discov Today2003;8:307-315.
11.
Vousden KH: Activation of the p53 tumor suppressor gene. Biochem Biophys Acta2002;1602:47-59.
12.
Chen L, Agrawal S, Zhou W, Zhang R, Chen J: Synergistic activation of p53 by inhibition of MDM2 expression and DNA damage. Proc Natl Acad Sci USA1998;95:195-200.
13.
Zhang Z, Li M, Wang H, Agrawal S, Zhang R: Antisense therapy targeting MDM2 oncogene in prostate cancer: effects on proliferation, apoptosis, multiple gene expression, and chemotherapy. Proc Natl Acad Sci USA2003;100:11636-11641.
14.
Chen L, Lu W, Agrawal S, Zhou W, Zhang R, Chen J: Ubiquitous induction of p53 in tumor cells by antisense inhibition of MDM2 expression. Mol Med1999;5:21-34.
15.
Fang S, Jensen JP, Ludwig RL, Vousden KH, Weissman AM: Mdm2 is a RING finger-dependent ubiquitin protein ligase for itself and p53. J Biol Chem2000;275:8945-8951.
16.
Honda R, Yasuda H: Association of p19 ARF with Mdm2 inhibits ubiquitin ligase activity ofMdm2for tumor suppressor p53. EMBOJ1999;18:22-27.
17.
Midgley CA, Desterro JM, Saville MK, et al: An N-terminal p14ARF peptide blocks Mdm2-dependent ubiquitination in vitro and can activate p53 in vivo. Oncogene2000;19:2312-2323.
18.
Zhang Y, Wolf GW, Bhat K, et al: Ribosomal protein L11 negatively regulates oncoprotein MDM2 and mediates a p53-dependent ribosomal-stress checkpoint pathway. Mol Cell Biol2003;23:8902-8912.
19.
Stott F, Bates SA, James M, et al: The alternative product from the human CDKN2A locus, p14ARF, participates in a regulatory feedback loop with p53 and MDM2. EMBO J1998;17:5001-5014.
20.
Lohrum MA, Ludwig RL, Kubbutat MHG, Hanlon M, Vousden KH: Regulation of HDM2 activity by the ribosomal proteinL11. CancerCell2003;3:577-587.
21.
Pomerantz J, Schreiber-AgusN, Liégeois NJ, et al: The Ink4a tumor suppressor gene product, p19Arf, interacts with MDM2 and neutralizes MDM2's inhibition of p53. Cell1998;92:713-723.
22.
Kamijo T, Weber JD, Zambetti G, Zindy F, Roussel MF, Sherr CJ: Functional and physical interactions of the ARF tumor suppressor with p53 and Mdm2. Proc Natl Acad Sci USA1998;95:8292-8297.
23.
Jensen JP, Bates PW, Yang M, Vierstra RD, Weissman AM: Identification of a family of closely related human ubiquitin conjugating enzymes. J Biol Chem1995;270:30408-30414.
24.
Hatakeyama S, Jensen JP, Weissman AM: Subcellular localization and ubiquitin-conjugating enzyme (E2) interactions of mammalian HECT family ubiquitin protein ligases. J Biol Chem1997;272:15085-15092.
25.
Yang Y, Fang S, Jensen JP, Weissman AM, Ashwell JD: Ubiquitin protein ligase activity of IAPs and their degradation in proteasomes in response to apoptotic stimuli. Science2000;288:874-877.
Honda R, Tanaka H, Yasuda H: Oncoprotein MDM2 is a ubiquitin ligase E3 for tumor suppressor p53. FEBS Lett1997;420:25-27.
28.
Sharp DA, Kratowicz SA, Sank MJ, George DL: Stabilization of the MDM2 oncogene by interaction with the structurally related MDMX protein. J Biol Chem1999;274:38189-38196.
29.
Badciong JC, Haas AL: MdmX is a RING finger ubiquitin ligase capable of synergistically enhancing Mdm2 ubiquitination. J Biol Chem2002;277:49668-49675.
30.
Jackson MW, Berberich SJ: MdmX protects p53 from Mdm2-mediated degradation. Mol Cell Biol2000;20:1001-1007.
31.
Stad R, Ramos YF, Little N, et al: HdmX stabilizes Mdm2 and p53. J Biol Chem2000;275:28039-28044.
32.
Meulmeester E, Frenk R, Stad R, et al: Critical role for a central part ofMdm2 in the ubiquitylation of p53. Mol Biol Cell2003;23:4929-4938.
33.
Fabbro D, Garcia-Echeverria C: Targeting protein kinases in cancer therapy. Curr Opin Drug Discov Dev2002;5:701-712.
34.
Lai Z, Yang T, Kim YB, et al: Differentiation of Hdm2-mediated p53 ubiquitination and Hdm2 autoubiquitination activity by small molecular weight inhibitors. Proc Natl Acad Sci USA2002;99:14734-14739.