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/ lu-2007a-1 The Wip1 Phosphatase Acts as a Gatekeeper in the p53-Mdm2 Autoregulatory Loop. human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Lu X., Ma O., Nguyen TA., Jones SN., Oren M., Donehower LA. Apoptosis - Interactions involving proteins with a function related to apoptosis xlu@biol.sc.edu,larryd@bcm.tmc.edu Cancer Cell (1535-6108) 2007 Only protein-protein interactions 8 imex curation Cancer - Interactions investigated in the context of cancer lu-2007a-2 The Wip1 Phosphatase Acts as a Gatekeeper in the p53-Mdm2 Autoregulatory Loop. in vitro In vitro phosphatase assay phosphatase assay predetermined predetermined participant predetermined Lu X., Ma O., Nguyen TA., Jones SN., Oren M., Donehower LA. Apoptosis - Interactions involving proteins with a function related to apoptosis xlu@biol.sc.edu,larryd@bcm.tmc.edu Cancer Cell (1535-6108) 2007 Only protein-protein interactions 8 imex curation Cancer - Interactions investigated in the context of cancer lu-2007a-3 The Wip1 Phosphatase Acts as a Gatekeeper in the p53-Mdm2 Autoregulatory Loop. human-saos-2 Homo sapiens SAOS-2 cell line established from primary osteogenic sarcoma saos-2 Homo sapiens SAOS-2 cell line established from primary osteogenic sarcoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Lu X., Ma O., Nguyen TA., Jones SN., Oren M., Donehower LA. Apoptosis - Interactions involving proteins with a function related to apoptosis p53 null Saos-2 cells were transfected with control scrambled siRNA, Wip1 expression vector DNA, or Wip1 siRNAs, together with the same amount of p53 expression vector DNA. Twenty-four hours after transfection, cells were treated with or without 5 Gy IR and protease inhibitors MG132 and MG102, and harvested 6 hr later. Cell lysates were immunoprecipitated with p53 antibody and immunoblotted with anti-ubiquitin antibody (left panel) or anti-Mdm2 xlu@biol.sc.edu,larryd@bcm.tmc.edu Cancer Cell (1535-6108) 2007 Only protein-protein interactions 8 imex curation Cancer - Interactions investigated in the context of cancer lu-2007a-4 The Wip1 Phosphatase Acts as a Gatekeeper in the p53-Mdm2 Autoregulatory Loop. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Lu X., Ma O., Nguyen TA., Jones SN., Oren M., Donehower LA. Apoptosis - Interactions involving proteins with a function related to apoptosis 293HEK cells that inducibly express Wip1 in the presence of doxycycline were transfected with equal amounts of HA-Mdm2 and FLAG-Mdm2 expression vector DNA, treated with or without 5 Gy IR, and harvested 2 hr later. The Mdm2 self-interaction was analyzed by immunoprecipitation and immunoblotting. xlu@biol.sc.edu,larryd@bcm.tmc.edu Cancer Cell (1535-6108) 2007 Only protein-protein interactions 8 imex curation Cancer - Interactions investigated in the context of cancer lu-2007a-5 The Wip1 Phosphatase Acts as a Gatekeeper in the p53-Mdm2 Autoregulatory Loop. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Lu X., Ma O., Nguyen TA., Jones SN., Oren M., Donehower LA. Apoptosis - Interactions involving proteins with a function related to apoptosis 293HEK cells were transfected with HA-ubiquitin expression vector, together with control DNA, or Wip1 expression vector DNA or Wip1 siRNA-1. Transfected cells were treated with or without 5 Gy IR and with protease inhibitor MG132, and harvested 2 hr after IR. Cell lysates were immunoprecipitated with anti-HA antibody and immunoblotted by anti-Mdm2 antibody. xlu@biol.sc.edu,larryd@bcm.tmc.edu Cancer Cell (1535-6108) 2007 Only protein-protein interactions 8 imex curation Cancer - Interactions investigated in the context of cancer ppm1d_human Protein phosphatase 1D Protein phosphatase 2C isoform delta p53-induced protein phosphatase 1 Protein phosphatase magnesium-dependent 1 delta PPM1D WIP1 protein protein human Homo sapiens Human MAGLYSLGVSVFSDQGGRKYMEDVTQIVVEPEPTAEEKPSPRRSLSQPLPPRPSPAALPGGEVSGKGPAVAAREARDPLPDAGASPAPSRCCRRRSSVAFFAVCDGHGGREAAQFAREHLWGFIKKQKGFTSSEPAKVCAAIRKGFLACHLAMWKKLAEWPKTMTGLPSTSGTTASVVIIRGMKMYVAHVGDSGVVLGIQDDPKDDFVRAVEVTQDHKPELPKERERIEGLGGSVMNKSGVNRVVWKRPRLTHNGPVRRSTVIDQIPFLAVARALGDLWSYDFFSGEFVVSPEPDTSVHTLDPQKHKYIILGSDGLWNMIPPQDAISMCQDQEEKKYLMGEHGQSCAKMLVNRALGRWRQRMLRADNTSAIVICISPEVDNQGNFTNEDELYLNLTDSPSYNSQETCVMTPSPCSTPPVKSLEEDPWPRVNSKDHIPALVRSNAFSENFLEVSAEIARENVQGVVIPSKDPEPLEENCAKALTLRIHDSLNNSLPIGLVPTNSTNTVMDQKNLKMSTPGQMKAQEIERTPPTNFKRTLEESNSGPLMKKHRRNGLSRSSGAQPASLPTTSQRKNSVKLTMRRRLRGQKKIGNPLLHQHRKTVCVC 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 ubp7_human Ubiquitin carboxyl-terminal hydrolase 7 USP7 HAUSP Ubiquitin thioesterase 7 Ubiquitin-specific-processing protease 7 Deubiquitinating enzyme 7 Herpesvirus-associated ubiquitin-specific protease protein protein human Homo sapiens Human MNHQQQQQQQKAGEQQLSEPEDMEMEAGDTDDPPRITQNPVINGNVALSDGHNTAEEDMEDDTSWRSEATFQFTVERFSRLSESVLSPPCFVRNLPWKIMVMPRFYPDRPHQKSVGFFLQCNAESDSTSWSCHAQAVLKIINYRDDEKSFSRRISHLFFHKENDWGFSNFMAWSEVTDPEKGFIDDDKVTFEVFVQADAPHGVAWDSKKHTGYVGLKNQGATCYMNSLLQTLFFTNQLRKAVYMMPTEGDDSSKSVPLALQRVFYELQHSDKPVGTKKLTKSFGWETLDSFMQHDVQELCRVLLDNVENKMKGTCVEGTIPKLFRGKMVSYIQCKEVDYRSDRREDYYDIQLSIKGKKNIFESFVDYVAVEQLDGDNKYDAGEHGLQEAEKGVKFLTLPPVLHLQLMRFMYDPQTDQNIKINDRFEFPEQLPLDEFLQKTDPKDPANYILHAVLVHSGDNHGGHYVVYLNPKGDGKWCKFDDDVVSRCTKEEAIEHNYGGHDDDLSVRHCTNAYMLVYIRESKLSEVLQAVTDHDIPQQLVERLQEEKRIEAQKRKERQEAHLYMQVQIVAEDQFCGHQGNDMYDEEKVKYTVFKVLKNSSLAEFVQSLSQTMGFPQDQIRLWPMQARSNGTKRPAMLDNEADGNKTMIELSDNENPWTIFLETVDPELAASGATLPKFDKDHDVMLFLKMYDPKTRSLNYCGHIYTPISCKIRDLLPVMCDRAGFIQDTSLILYEEVKPNLTERIQDYDVSLDKALDELMDGDIIVFQKDDPENDNSELPTAKEYFRDLYHRVDVIFCDKTIPNDPGFVVTLSNRMNYFQVAKTVAQRLNTDPMLLQFFKSQGYRDGPGNPLRHNYEGTLRDLLQFFKPRQPKKLYYQQLKMKITDFENRRSFKCIWLNSQFREEEITLYPDKHGCVRDLLEECKKAVELGEKASGKLRLLEIVSYKIIGVHQEDELLECLSPATSRTFRIEEIPLDQVDIDKENEMLVTVAHFHKEVFGTFGIPFLLRIHQGEHFREVMKRIQSLLDIQEKEFEKFKFAIVMMGRHQYINEDEYEVNLKDFEPQPGNMSHPRPWLGLDHFNKAPKRSRYTYLEKAIKIHN p53_human Cellular tumor antigen p53 TP53 P53 Tumor suppressor p53 Phosphoprotein p53 Antigen NY-CO-13 protein protein human Homo sapiens Human MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLSPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYKQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPPEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMFRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD ubb_human Polyubiquitin-B UBB protein protein human Homo sapiens Human MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGC ppm1d-mdm2 Endogenous Wip1 coimmunoprecipitates endogenous Mdm2 in U2OS lysates 677375 N/A 677380 unspecified role unspecified role bait bait N/A 677381 unspecified role unspecified role prey prey physical association physical association false false false 3 A mdm2-ppm1d Endogenous Wip1 coimmunoprecipitates with endogenous Mdm2 in U2OS lysates 677375 N/A 677381 unspecified role unspecified role bait bait N/A 677380 unspecified role unspecified role prey prey physical association physical association false false false 3 A Transfection with Wip1 siRNA abolishes this interaction due to the absence of Wip1. usp7-mdm2-6 Wip1 enhances the HAUSP-Mdm2 interaction. 677375 N/A 677382 unspecified role unspecified role bait bait N/A 677381 unspecified role unspecified role prey prey physical association physical association false false false 6 C Wip1 - Overexpression of Wip1 enhances the HAUSP-Mdm2 interaction. ppm1d-mdm2-1 Wip1 dephosphorylates Mdm2 phosphopeptide on Ser-395 in vitro 677376 N/A 677381 enzyme target enzyme target substrate substrate unspecified role unspecified role region opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 677380 enzyme enzyme unspecified role unspecified role region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. dephosphorylation dephosphorylation reaction dephosphorylation false false false The Mdm2 phosphopeptide used was Ac-ESEDYpSQPSTS-amide. 3 C, 3 F The Mdm2 phosphopeptide was dephosphorylated on Ser395. Wip1 is insensitive to okadaic acid. Mg2+ - Wip1 phosphatase activity is Mg2+-dependent. PP1, PP2A and PP2Calpha did not dephosphorylate Mdm2 serine 395 phosphopeptide. ppm1d-mdm2-2 Wip1 dephosphorylates intact Mdm2 on Ser-395 in vitro 677376 N/A 677381 enzyme target enzyme target substrate substrate unspecified role unspecified role region opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain false human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference N/A 677380 enzyme enzyme unspecified role unspecified role region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. dephosphorylation dephosphorylation reaction dephosphorylation false false false 3 D Mdm2 was dephosphorylated on Ser395. Mg2+ - Wip1 phosphatase activity is Mg2+-dependent. tp53-rps27a-1 Wip1 promotes p53 ubiquitination in Saos-2 cells 677377 N/A 677383 unspecified role unspecified role bait bait N/A 677384 unspecified role unspecified role prey prey physical association physical association false false false 2 B P04637 transfection Overexpressed Wip1 resulted in increased p53 ubiquitination, while reduction of Wip1 via Wip1 siRNA suppressed p53 ubiquitination in both IR-treated and untreated cells. mdm2-mdm2 Wip1 diminishes Mdm2 self-interaction. 677378 N/A 677381 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 677381 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false 6 A Increased Wip1 expression reduced the interaction between HA-Mdm2 and FLAG-Mdm2 before and after IR treatment. Q00987 transfection rsp27a-mdm2 Ubiquitination of Mdm2 is inhibited by Wip1. 677379 N/A 677384 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 677381 unspecified role unspecified role prey prey physical association physical association false false false 6 B Wip1 - This interaction is inhibited by Wip1. Ionizing radiation (IR) - This interaction is enhanced by IR treatment.