IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ http://www.ebi.ac.uk/ macias-2010-1 An ARF-independent c-MYC-activated tumor suppression pathway mediated by ribosomal protein-Mdm2 Interaction. mouse-embryonic_fibroblast Mus musculus embryonic fibroblasts embryonic_fibroblast Embryonic fibroblast Fibroblasts taken from an embryonic as opposed to a foetal or adult source. Foetus is defined as the stage at which tissue diferentiation is initiated. Tissue taken from an embryonic as opposed to a foetal or adult source. Embryo is defined as the development stage prior to organ formation. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Macias E., Jin A., Deisenroth C., Bhat K., Mao H., Lindstrom MS., Zhang Y. Apoptosis - Interactions involving proteins with a function related to apoptosis The Mdm2 C305F mutation has been remapped to cys306phe MEFs were isolated at embryonic day 13.5 (E13.5). Amino acid sequence numbering for Mouse MDM2 exons 7-12 was obtained from Iwakuma and Lozano, Molecular Cancer Research, 2003. ypzhang@med.unc.edu Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer macias-2010-2 An ARF-independent c-MYC-activated tumor suppression pathway mediated by ribosomal protein-Mdm2 Interaction. mouse-spleen Mus musculus spleen spleen spleen anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Macias E., Jin A., Deisenroth C., Bhat K., Mao H., Lindstrom MS., Zhang Y. Apoptosis - Interactions involving proteins with a function related to apoptosis The Mdm2 C305F mutation has been remapped to cys306phe MEFs were isolated at embryonic day 13.5 (E13.5). Amino acid sequence numbering for Mouse MDM2 exons 7-12 was obtained from Iwakuma and Lozano, Molecular Cancer Research, 2003. ypzhang@med.unc.edu Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer arf_mouse Tumor suppressor ARF Cdkn2a p19ARF Alternative reading frame Cyclin-dependent kinase inhibitor 2A protein protein mouse Mus musculus Mouse MGRRFLVTVRIQRAGRPLQERVFLVKFVRSRRPRTASCALAFVNMLLRLERILRRGPHRNPGPGDDDGQRSRSSSSAQLRCRFELRGPHYLLPPGARRSAGRLPGHAGGAARVRGSAGCARCLGSPAARLGPRAGTSRHRAIFAFRWVLFVFRWVVFVYRWERRPDRRA rl23_mouse 60S ribosomal protein L23 Rpl23 protein protein mouse Mus musculus Mouse MSKRGRGGSSGAKFRISLGLPVGAVINCADNTGAKNLYIISVKGIKGRLNRLPAAGVGDMVMATVKKGKPELRKKVHPAVVIRQRKSYRRKDGVFLYFEDNAGVIVNNKGEMKGSAITGPVAKECADLWPRIASNAGSIA rl5_mouse 60S ribosomal protein L5 Rpl5 protein protein mouse Mus musculus Mouse MGFVKVVKNKAYFKRYQVRFRRRREGKTDYYARKRLVIQDKNKYNTPKYRMIVRVTNRDIICQIAYARIEGDMIVCAAYAHELPKYGVKVGLTNYAAAYCTGLLLARRLLNRFGMDKIYEGQVEVNGGEYNVESIDGQPGAFTCYLDAGLARTTTGNKVFGALKGAVDGGLSIPHSTKRFPGYDSESKEFNAEVHRKHIMGQNVADYMRYLMEEDEDAYKKQFSQYIKNNVTPDMMEEMYKKAHAAIRENPVYEKKPKREVKKKRWNRPKMSLAQKKDRVAQKKASFLRAQERAAES p53_mouse Cellular tumor antigen p53 Tp53 P53 Trp53 Tumor suppressor p53 protein protein mouse Mus musculus Mouse MTAMEESQSDISLELPLSQETFSGLWKLLPPEDILPSPHCMDDLLLPQDVEEFFEGPSEALRVSGAPAAQDPVTETPGPVAPAPATPWPLSSFVPSQKTYQGNYGFHLGFLQSGTAKSVMCTYSPPLNKLFCQLAKTCPVQLWVSATPPAGSRVRAMAIYKKSQHMTEVVRRCPHHERCSDGDGLAPPQHLIRVEGNLYPEYLEDRQTFRHSVVVPYEPPEAGSEYTTIHYKYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRDSFEVRVCACPGRDRRTEEENFRKKEVLCPELPPGSAKRALPTCTSASPPQKKKPLDGEYFTLKIRGRKRFEMFRELNEALELKDAHATEESGDSRAHSSYLKTKKGQSTSRHKKTMVKKVGPDSD rl11_mouse 60S ribosomal protein L11 Rpl11 protein protein mouse Mus musculus Mouse MAQDQGEKENPMRELRIRKLCLNICVGESGDRLTRAAKVLEQLTGQTPVFSKARYTVRSFGIRRNEKIAVHCTVRGAKAEEILEKGLKVREYELRKNNFSDTGNFGFGIQEHIDLGIKYDPSIGIYGLDFYVVLGRPGFSIADKKRRTGCIGAKHRISKEEAMRWFQQKYDGIILPGK mdm2_mouse E3 ubiquitin-protein ligase Mdm2 p53-binding protein Mdm2 Oncoprotein Mdm2 Double minute 2 protein Mdm2 RING-type E3 ubiquitin transferase Mdm2 protein protein mouse Mus musculus Mouse MCNTNMSVSTEGAASTSQIPASEQETLVRPKPLLLKLLKSVGAQNDTYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVAVSQQDSGTSLSESRRQPEGGSDLKDPLQAPPEEKPSSSDLISRLSTSSRRRSISETEENTDELPGERHRKRRRSLSFDPSLGLCELREMCSGGSSSSSSSSSESTETPSHQDLDDGVSEHSGDCLDQDSVSDQFSVEFEVESLDSEDYSLSDEGHELSDEDDEVYRVTVYQTGESDTDSFEGDPEISLADYWKCTSCNEMNPPLPSHCKRCWTLRENWLPDDKGKDKVEISEKAKLENSAQAEEGLDVPDGKKLTENDAKEPCAEEDSEEKAEQTPLSQESDDYSQPSTSSSIVYSSQESVKELKEETQDKDESVESSFSLNAIEPCVICQGRPKNGCIVHGKTGHLMSCFTCAKKLKKRNKPCPVCRQPIQMIVLTYFN mdm2-wt 986357 n/a 986359 unspecified role unspecified role prey prey n/a 986360 unspecified role unspecified role prey prey n/a 986361 unspecified role unspecified role prey prey n/a 986362 unspecified role unspecified role prey prey n/a 986363 unspecified role unspecified role prey prey n/a 986364 unspecified role unspecified role bait bait association association false false false Fig 2A, 6E. Mutant Mdm2 (cys306phe) did not interact with the L5 protein and had a weak interaction with the L11 protein. Actinomycin D Mutant Mdm2 (minus N terminal amino acids) does bind to p53 in Fig 2A, yet it does NOT bind p53 as shown in Fig 6E. In both Mdm2 mutant and p19 Arf knockout cell lines the addition of Actinomycin D increased the interaction of Mdm2 with L11 and p19Arf. In Fig 6E no interaction between Mdm2(wt) and p53 was observed in the absence of Act D, and no interaction between p53 and the mutant Mdm2 was noted at all. rl23_mdm2 986357 n/a 986360 unspecified role unspecified role bait bait n/a 986364 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind range range certain certain sequence position certain false P23804:p.Cys306Phe mutation mutation range range range range false physical association physical association false false false Fig 2A, 2B. Mdm2C306F failed to bind L5 and L11, but was able to bind L23 and p53. mdm2-wt-1 986358 n/a 986363 unspecified role unspecified role prey prey n/a 986364 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind range range certain certain sequence position certain false P23804:p.Cys306Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false The L11 in Eu-Myc;Mdm2C306F/C306F splenocytes, even though expressed to a higher level than in Eu-Myc;Mdm2+/+ splenocytes, showed very little interactions with Mdm2. Fig 5E.