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/ Negative regulation of p53 functions by Daxx and the involvement of MDM2. Negative regulation of p53 functions by Daxx and the involvement of MDM2. J. Biol. Chem. (0021-9258) 2004 imex curation Zhao LY., Liu J., Sidhu GS., Niu Y., Liu Y., Wang R., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer p53-null HCT116 cells were transfected with various combinations of expression plasmids. The authors confirmed by email that the species of origin of the MDM2 construct used was human. dliao@ufl.edu 01-Feb-2008: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zhao LY., Liu J., Sidhu GS., Niu Y., Liu Y., Wang R., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis p53-null HCT116 cells were transfected with various combinations of expression plasmids. The authors confirmed by email that the species of origin of the MDM2 construct used was human. dliao@ufl.edu Accepted 2008-FEB-10 by ORCHARD. 01-Feb-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2004 orchard Only protein-protein interactions imex curation Negative regulation of p53 functions by Daxx and the involvement of MDM2. Negative regulation of p53 functions by Daxx and the involvement of MDM2. J. Biol. Chem. (0021-9258) 2004 imex curation Zhao LY., Liu J., Sidhu GS., Niu Y., Liu Y., Wang R., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer p53-null HCT116 cells were transfected with various combinations of expression plasmids. The authors confirmed by email that the species of origin of the MDM2 construct used was human. dliao@ufl.edu 01-Feb-2008: Contacted by JYOTI. orchard Only protein-protein interactions human-hct116 Human colon carcinoma HCT116 cells hct116 Human colon carcinoma line HCT116 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zhao LY., Liu J., Sidhu GS., Niu Y., Liu Y., Wang R., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis p53-null HCT116 cells were transfected with various combinations of expression plasmids. The authors confirmed by email that the species of origin of the MDM2 construct used was human. dliao@ufl.edu Accepted 2008-FEB-10 by ORCHARD. 01-Feb-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2004 orchard Only protein-protein interactions imex curation Negative regulation of p53 functions by Daxx and the involvement of MDM2. Negative regulation of p53 functions by Daxx and the involvement of MDM2. J. Biol. Chem. (0021-9258) 2004 imex curation Zhao LY., Liu J., Sidhu GS., Niu Y., Liu Y., Wang R., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer p53-null HCT116 cells were transfected with various combinations of expression plasmids. The authors confirmed by email that the species of origin of the MDM2 construct used was human. dliao@ufl.edu 01-Feb-2008: Contacted by JYOTI. orchard Only protein-protein interactions 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 tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Zhao LY., Liu J., Sidhu GS., Niu Y., Liu Y., Wang R., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis p53-null HCT116 cells were transfected with various combinations of expression plasmids. The authors confirmed by email that the species of origin of the MDM2 construct used was human. dliao@ufl.edu Accepted 2008-FEB-10 by ORCHARD. 01-Feb-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2004 orchard Only protein-protein interactions imex curation Negative regulation of p53 functions by Daxx and the involvement of MDM2. Negative regulation of p53 functions by Daxx and the involvement of MDM2. J. Biol. Chem. (0021-9258) 2004 imex curation Zhao LY., Liu J., Sidhu GS., Niu Y., Liu Y., Wang R., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer p53-null HCT116 cells were transfected with various combinations of expression plasmids. The authors confirmed by email that the species of origin of the MDM2 construct used was human. dliao@ufl.edu 01-Feb-2008: Contacted by JYOTI. orchard Only protein-protein interactions 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. fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Zhao LY., Liu J., Sidhu GS., Niu Y., Liu Y., Wang R., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis p53-null HCT116 cells were transfected with various combinations of expression plasmids. The authors confirmed by email that the species of origin of the MDM2 construct used was human. dliao@ufl.edu Accepted 2008-FEB-10 by ORCHARD. 01-Feb-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2004 orchard Only protein-protein interactions imex curation Negative regulation of p53 functions by Daxx and the involvement of MDM2. Negative regulation of p53 functions by Daxx and the involvement of MDM2. J. Biol. Chem. (0021-9258) 2004 imex curation Zhao LY., Liu J., Sidhu GS., Niu Y., Liu Y., Wang R., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer p53-null HCT116 cells were transfected with various combinations of expression plasmids. The authors confirmed by email that the species of origin of the MDM2 construct used was human. dliao@ufl.edu 01-Feb-2008: Contacted by JYOTI. orchard Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Zhao LY., Liu J., Sidhu GS., Niu Y., Liu Y., Wang R., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis p53-null HCT116 cells were transfected with various combinations of expression plasmids. The authors confirmed by email that the species of origin of the MDM2 construct used was human. dliao@ufl.edu Accepted 2008-FEB-10 by ORCHARD. 01-Feb-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2004 orchard Only protein-protein interactions imex curation Negative regulation of p53 functions by Daxx and the involvement of MDM2. Negative regulation of p53 functions by Daxx and the involvement of MDM2. J. Biol. Chem. (0021-9258) 2004 imex curation Zhao LY., Liu J., Sidhu GS., Niu Y., Liu Y., Wang R., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer p53-null HCT116 cells were transfected with various combinations of expression plasmids. The authors confirmed by email that the species of origin of the MDM2 construct used was human. dliao@ufl.edu 01-Feb-2008: Contacted by JYOTI. orchard Only protein-protein interactions human-hct116 Human colon carcinoma HCT116 cells hct116 Human colon carcinoma line HCT116 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Zhao LY., Liu J., Sidhu GS., Niu Y., Liu Y., Wang R., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis p53-null HCT116 cells were transfected with various combinations of expression plasmids. The authors confirmed by email that the species of origin of the MDM2 construct used was human. dliao@ufl.edu Accepted 2008-FEB-10 by ORCHARD. 01-Feb-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2004 orchard Only protein-protein interactions imex curation mdm2_human E3 ubiquitin-protein ligase Mdm2 MDM2 p53-binding protein Mdm2 Oncoprotein Mdm2 Double minute 2 protein RING-type E3 ubiquitin transferase Mdm2 protein protein human Homo sapiens Human MCNTNMSVPTDGAVTTSQIPASEQETLVRPKPLLLKLLKSVGAQKDTYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSENRCHLEGGSDQKDLVQELQEEKPSSSHLVSRPSTSSRRRAISETEENSDELSGERQRKRHKSDSISLSFDESLALCVIREICCERSSSSESTGTPSNPDLDAGVSEHSGDWLDQDSVSDQFSVEFEVESLDSEDYSLSEEGQELSDEDDEVYQVTVYQAGESDTDSFEEDPEISLADYWKCTSCNEMNPPLPSHCNRCWALRENWLPEDKGKDKGEISEKAKLENSTQAEEGFDVPDCKKTIVNDSRESCVEENDDKITQASQSQESEDYSQPSTSSSIIYSSQEDVKEFEREETQDKEESVESSLPLNAIEPCVICQGRPKNGCIVHGKTGHLMACFTCAKKLKKRNKPCPVCRQPIQMIVLTYFP DIP protein Q00987 original sequence version: 93 DIP protein Q00987-1 original sequence version: 192 DIP protein Q00987 original sequence version: 88 DIP protein Q00987 original sequence version: 180 DIP protein Q00987 original sequence version: 188 DIP protein Q00987 original sequence version: 189 DIP protein Q00987 original sequence version: 115 DIP protein Q00987-1 original sequence version: null DIP protein Q00987 original sequence version: 149 DIP protein Q00987 original sequence version: 146 DIP protein Q00987 original sequence version: 140 DIP protein Q00987 original sequence version: 183 DIP protein Q00987 original sequence version: 145 DIP protein Q00987 original sequence version: 182 DIP protein Q00987 original sequence version: 151 DIP protein Q00987 original sequence version: 156 DIP protein Q00987-1 original sequence version: 160 DIP protein Q00987 original sequence version: 160 DIP protein Q00987 original sequence version: 168 DIP protein Q00987 original sequence version: 186 DIP protein Q00987 original sequence version: 206 F37CE163876BC983 daxx_human Death domain-associated protein 6 DAXX BING2 DAP6 Daxx Fas death domain-associated protein ETS1-associated protein 1 protein protein human Homo sapiens Human MATANSIIVLDDDDEDEAAAQPGPSHPLPNAASPGAEAPSSSEPHGARGSSSSGGKKCYKLENEKLFEEFLELCKMQTADHPEVVPFLYNRQQRAHSLFLASAEFCNILSRVLSRARSRPAKLYVYINELCTVLKAHSAKKKLNLAPAATTSNEPSGNNPPTHLSLDPTNAENTASQSPRTRGSRRQIQRLEQLLALYVAEIRRLQEKELDLSELDDPDSAYLQEARLKRKLIRLFGRLCELKDCSSLTGRVIEQRIPYRGTRYPEVNRRIERLINKPGPDTFPDYGDVLRAVEKAAARHSLGLPRQQLQLMAQDAFRDVGIRLQERRHLDLIYNFGCHLTDDYRPGVDPALSDPVLARRLRENRSLAMSRLDEVISKYAMLQDKSEEGERKKRRARLQGTSSHSADTPEASLDSGEGPSGMASQGCPSASRAETDDEDDEESDEEEEEEEEEEEEEATDSEEEEDLEQMQEGQEDDEEEDEEEEAAAGKDGDKSPMSSLQISNEKNLEPGKQISRSSGEQQNKGRIVSPSLLSEEPLAPSSIDAESNGEQPEELTLEEESPVSQLFELEIEALPLDTPSSVETDISSSRKQSEEPFTTVLENGAGMVSSTSFNGGVSPHNWGDSGPPCKKSRKEKKQTGSGPLGNSYVERQRSVHEKNGKKICTLPSPPSPLASLAPVADSSTRVDSPSHGLVTSSLCIPSPARLSQTPHSQPPRPGTCKTSVATQCDPEEIIVLSDSD 1B309ADDAA878040 p53_human Cellular tumor antigen p53 TP53 P53 Tumor suppressor p53 Phosphoprotein p53 Antigen NY-CO-13 protein protein human Homo sapiens Human MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLSPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYKQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPPEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMFRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD DIP protein P04637 original sequence version: 121 DIP protein P04637 original sequence version: 115 DIP protein P04637-1 original sequence version: 224 DIP protein P04637 original sequence version: 117 DIP protein P04637-1 original sequence version: 226 DIP protein P04637 original sequence version: 226 DIP protein P04637 original sequence version: 212 DIP protein P04637 original sequence version: 220 DIP protein P04637-1 original sequence version: 220 DIP protein P04637-1 original sequence version: 145 DIP protein P04637 original sequence version: 146 DIP protein P04637 original sequence version: 145 DIP protein P04637 original sequence version: 166 DIP protein P04637-1 original sequence version: 166 DIP protein P04637 original sequence version: 169 DIP protein P04637 original sequence version: 177 DIP protein P04637 original sequence version: 175 DIP protein P04637-1 original sequence version: 175 DIP protein P04637-1 original sequence version: 172 DIP protein P04637-1 original sequence version: 169 DIP protein P04637 original sequence version: 170 DIP protein P04637 original sequence version: 172 DIP protein P04637-1 original sequence version: 194 DIP protein P04637 original sequence version: 196 DIP protein P04637 original sequence version: 185 DIP protein P04637-1 original sequence version: 191 DIP protein P04637 original sequence version: 191 DIP protein P04637 original sequence version: 180 DIP protein P04637-1 original sequence version: 197 DIP protein P04637 original sequence version: 183 DIP protein P04637-1 original sequence version: 183 DIP protein P04637-1 original sequence version: 185 DIP protein P04637 original sequence version: 199 DIP protein P04637 original sequence version: 201 DIP protein P04637 original sequence version: 189 DIP protein P04637 original sequence version: 235 DIP protein P04637 original sequence version: 230 DIP protein P04637 original sequence version: 234 DIP protein P04637-1 original sequence version: 234 DIP protein P04637-1 original sequence version: 235 DIP protein P04637 original sequence version: 239 DIP protein P04637-1 original sequence version: 212 DIP protein P04637-1 original sequence version: 213 DIP protein P04637 original sequence version: 213 DIP protein P04637 original sequence version: 206 DIP protein P04637-1 original sequence version: 188 DIP protein P04637-1 original sequence version: 228 DIP protein P04637 original sequence version: 208 DIP protein P04637 original sequence version: 240 DIP protein P04637-1 original sequence version: 240 DIP protein P04637 original sequence version: 204 DIP protein P04637-1 original sequence version: 204 DIP protein P04637-1 original sequence version: 209 DIP protein P04637-1 original sequence version: 203 DIP protein P04637 original sequence version: 221 DIP protein P04637-1 original sequence version: 152 DIP protein P04637 original sequence version: 217 DIP protein P04637-1 original sequence version: 140 DIP protein P04637 original sequence version: 215 DIP protein P04637 original sequence version: 214 DIP protein P04637-1 original sequence version: 182 DIP protein P04637 original sequence version: 182 DIP protein P04637 original sequence version: 188 DIP protein P04637 original sequence version: 192 DIP protein P04637-1 original sequence version: 192 DIP protein P04637 original sequence version: 218 DIP protein P04637-1 original sequence version: 218 AD5C149FD8106131 tp53-daxx 1 8 unspecified role unspecified role prey prey spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 9 unspecified role unspecified role bait bait direct interaction direct interaction 1 A When the extract of insect Sf9 cells expressing Daxx was mixed with purified WT p53, Daxx was precipitated along with p53 in an IP assay using DO-1 daxx-tp53 1 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection direct interaction direct interaction 1 A When the extract of insect Sf9 cells expressing Daxx was mixed with purified WT p53, Daxx was precipitated along with p53 in an IP assay using DO-1 (Fig. 1A, lane 3). Reciprocally, p53 was precipitated from the same mixture using anti-Daxx antibody M112 daxx-tp53-1 5 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association 1 B, C Deletion of the acidic domain (aa 434-496) from Daxx abolished its interaction with p53 This indicates that the Daxx acidic domain (aa 434-496) is required for binding with p53. tp53-daxx-1 5 9 unspecified role unspecified role bait bait P04637:p.Arg248Trp mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R W Wed May 02 16:27:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg248trp' Tue Jun 30 17:06:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg248Trp' c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain P04637:p.[Lys370Ala;Lys372Ala;Lys373Ala;Lys381Ala;Lys382Ala] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 18:05:59 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys370ala,lys372ala,lys373ala,lys381ala,lys382ala' Tue Jun 30 18:46:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys370Ala;Lys372Ala;Lys373Ala;Lys381Ala;Lys382Ala]' P04637:p.Lys386Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K R Wed May 02 17:06:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys386arg' Tue Jun 30 18:02:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys386Arg' P04637:p.Lys386Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K E Wed May 02 20:27:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys386glu' Tue Jun 30 15:48:04 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys386Glu' P04637:p.Lys319_Lys321delinsAlaAlaAla mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain KKK AAA Wed May 02 14:27:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys_lys_lys319-321ala_ala_ala' Tue Jun 30 19:58:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys319_Lys321delinsAlaAlaAla' P04637:p.Lys319_Lys321delinsGluGluGlu mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain KKK EEE Wed May 02 17:09:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys_lys_lys319-321glu_glu_glu' Tue Jun 30 18:01:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys319_Lys321delinsGluGluGlu' P04637:p.[Lys372Ala;Lys373Ala] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 15:47:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys372ala,lys373ala' Tue Jun 30 18:09:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys372Ala;Lys373Ala]' P04637:p.[Lys372Glu;Lys373Glu] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K E certain certain sequence position certain certain certain sequence position certain K E Wed May 02 18:06:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys372glu,lys373glu' Tue Jun 30 20:14:41 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys372Glu;Lys373Glu]' P04637:p.[Lys381Ala;Lys382Ala] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 19:01:09 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys381ala,lys382ala' Tue Jun 30 18:45:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys381Ala;Lys382Ala]' P04637:p.[Lys381Glu;Lys382Glu] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K E certain certain sequence position certain certain certain sequence position certain K E Wed May 02 15:31:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys381glu,lys382glu' Tue Jun 30 17:19:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys381Glu;Lys382Glu]' P04637:p.[Lys370Ala;Lys372Ala;Lys373Ala] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 14:38:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys370ala,lys372ala,lys373ala' Tue Jun 30 20:16:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys370Ala;Lys372Ala;Lys373Ala]' P04637:p.[Lys370Glu;Lys372Glu;Lys373Glu] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K E certain certain sequence position certain certain certain sequence position certain K E certain certain sequence position certain certain certain sequence position certain K E Wed May 02 15:23:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys370glu,lys372glu,lys373glu' Tue Jun 30 17:30:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys370Glu;Lys372Glu;Lys373Glu]' P04637:p.[Lys372Glu;Lys373Glu;Lys381Glu;Lys382Glu] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K E certain certain sequence position certain certain certain sequence position certain K E certain certain sequence position certain certain certain sequence position certain K E certain certain sequence position certain certain certain sequence position certain K E Wed May 02 19:34:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys372glu,lys373glu,lys381glu,lys382glu' Tue Jun 30 19:30:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys372Glu;Lys373Glu;Lys381Glu;Lys382Glu]' P04637:p.Ser376Glu mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S E Wed May 02 15:19:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser376glu' Tue Jun 30 19:51:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser376Glu' P04637:p.[Ser376Glu;Ser392Glu] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S E certain certain sequence position certain certain certain sequence position certain S E Wed May 02 18:47:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser376glu,ser392glu' Tue Jun 30 15:52:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser376Glu;Ser392Glu]' P04637:p.[Lys319Arg;Lys320Arg;Lys321Arg;Lys372Arg;Lys373Arg;Lys381Arg;Lys382Arg] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K R certain certain sequence position certain certain certain sequence position certain K R certain certain sequence position certain certain certain sequence position certain K R certain certain sequence position certain certain certain sequence position certain K R certain certain sequence position certain certain certain sequence position certain K R certain certain sequence position certain certain certain sequence position certain K R certain certain sequence position certain certain certain sequence position certain K R Wed May 02 17:02:06 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys319arg,lys320arg,lys321arg,lys372arg,lys373arg,lys381arg,lys382arg' Tue Jun 30 19:52:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys319Arg;Lys320Arg;Lys321Arg;Lys372Arg;Lys373Arg;Lys381Arg;Lys382Arg]' 8 unspecified role unspecified role prey prey physical association physical association 1 D, E, F, G p53 constructs with progressive deletions from the NH2 terminus up to residue 253 were able to bind to Daxx Point mutations in the NH2 terminus of p53 did not affect binding to Daxx and nor did mutations in the core domain. However, C-terminal truncations of p53 abolished this interaction (aa 1-315 and 1-355), indicating the C-terminal regulatory domain is required for this interaction. daxx-tp53-2 6 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain association association 2 B, 3 B, C MDM2 - This interaction only occurred when cells were tansfected with MDM2. Q9UER7; P04637; Q00987 transfection Daxx constructs aa 1-574, 1-501, and a construct with deletion of aa 191-242 (a putative amphipathic helix) bound to p53 less effectively than wild-type Daxx. In addition, the Daxx construct with a deletion between aa 502 and 574 exhibited weak binding to p53. The Daxx NH2-terminal domain (aa 1-130) and the COOH-terminal constructs (aa 516-740 and 573-740) did not bind to p53. These results indicate that the acidic domain of Daxx is required for binding to p53. More MDM2 was precipitated with WT Daxx than with the Daxx fragments (aa 1-574 and 1-502), in parallel with the amounts of precipitated p53. Interestingly, although p53 was precipitated with Daxx Delta191-242 and Delta502-574 constructs, little MDM2 was coprecipitated with them. These results suggest that MDM2 may mediate the Daxx-p53 interaction, as a physical bridge and stabilizer of the complex, and by inhbiting covalent modifications of p53. Or, MDM2 may alter the conformation of Daxx, so exposing its p53-binding domain. Daxx constructs aa 1-574, 1-501, and a construct with deletion of aa 191-242 (a putative amphipathic helix (18)) bound to p53 less effectively than wild-type Daxx (lanes 4, 6, and 10). In addition, the Daxx construct with a deletion between aa 502 and 574 exhibited weak binding to p53 (Fig. 3B, lane 16). The Daxx NH2-terminal domain (aa 1-130) and the COOH-terminal constructs (aa 516-740 and 573-740) did not bind to p53. These results indicate that the acidic domain of Daxx is required for binding to p53, consistent with the results obtained from yeast two-hybrid assays (Fig. 1). To examine whether MDM2 was present in the same FLAGDaxx immunocomplexes, the same membrane shown in Fig. 3B was stripped and reprobed with anti-MDM2 antibody. MDM2 was coprecipitated with WT Daxx and its fragments (aa 1-574 and 1-502, Fig. 3C, lanes 2, 4, and 6). tp53-daxx-2 2 8 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role bait bait P04637:p.[Lys381Ala;Lys382Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 19:26:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys381ala,lys382ala' Tue Jun 30 18:31:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys381Ala;Lys382Ala]' P04637:p.[Lys381Arg;Lys382Arg] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K R certain certain sequence position certain certain certain sequence position certain K R Wed May 02 19:37:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys381arg,lys382arg' Tue Jun 30 17:36:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys381Arg;Lys382Arg]' P04637:p.Ser376Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S A Wed May 02 17:44:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser376ala' Tue Jun 30 19:04:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser376Ala' P04637:p.Ser376Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S E Wed May 02 16:56:16 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser376glu' Tue Jun 30 16:49:28 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser376Glu' P04637:p.[Lys381Glu;Lys382Glu] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K E certain certain sequence position certain certain certain sequence position certain K E Wed May 02 17:03:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys381glu,lys382glu' Tue Jun 30 20:02:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys381Glu;Lys382Glu]' P04637:p.[Leu22Glu;Trp23Ser] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L E certain certain sequence position certain certain certain sequence position certain W S Wed May 02 19:01:52 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu22glu,trp23ser' Tue Jun 30 16:24:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu22Glu;Trp23Ser]' physical association physical association 4 B Daxx was coprecipitated with WT p53 (Fig. 4B, lane 2) and to a lesser extent with p53 K381A/K382A (lane 6), K381R/K382R (lane 8), S376A (lane 10), and p53 S376E Q9UER7; P04637 transfection daxx-tp53-3 3 9 unspecified role unspecified role prey prey P04637:p.[Leu22Gln;Trp23Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L Q certain certain sequence position certain certain certain sequence position certain W S Wed May 02 16:55:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu22gln,trp23ser' Tue Jun 30 17:35:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu22Gln;Trp23Ser]' P04637:p.[Lys381Ala;Lys382Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 18:07:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys381ala,lys382ala' Tue Jun 30 15:47:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys381Ala;Lys382Ala]' 8 unspecified role unspecified role bait bait n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association 5 C WT p53 was very efficiently precipitated in the presence of MDM2 (Fig. 5C, lane 2). Small but detectable amounts of p53 was precipitated in the absence of MDM2 expression (lane 4, p53 band was visible upon longer exposure). p53 mutants K381A/K382A and L22Q/W23S were also coprecipitated with FLAG-Daxx, although to a much lesser extent than WT p53 MDM2 - Cotransfection with MDM2 significantly enhanced this interaction. Q9UER7; P04637 transfection daxx-mdm2-3 3 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association 5 D MDM2 appears to constitutively associate with Daxx p53 - In the absence of p53 expression the amount of coprecipitated MDM2 was slightly reduced. Q9UER7; Q00987 transfection daxx-tp53-4 4 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization 5 E Codistribution of Daxx, MDM2, and p53 in DKO MEF Daxx appeared in nuclear body-like structures as well as in the nucleolus, whereas both p53 and MDM2 were largely excluded from these locations. The immunostaining patterns of p53 or MDM2 were essentially the same, whether both together or each individually was coexpressed with GFP-Daxx. daxx-mdm2-4 4 8 unspecified role unspecified role bait bait region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey colocalization colocalization 5 E Codistribution of Daxx, MDM2, and p53 in DKO MEF Daxx appeared in nuclear body-like structures as well as in the nucleolus, whereas both p53 and MDM2 were largely excluded from these locations. The immunostaining patterns of p53 or MDM2 were essentially the same, whether both together or each individually was coexpressed with GFP-Daxx.