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/ Acetylation is indispensable for p53 activation. Acetylation is indispensable for p53 activation. Cell (0092-8674) 2008 imex curation Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Change to acetylation assay when CVs update Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 7 in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Change to acetylation assay when CVs update Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu Accepted 2008-OCT-29 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 7 imex curation Acetylation is indispensable for p53 activation. Acetylation is indispensable for p53 activation. Cell (0092-8674) 2008 imex curation Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Change to acetylation assay when CVs update Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 7 in vitro In vitro acetylase assay acetylase assay predetermined predetermined participant predetermined Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu Accepted 2008-OCT-29 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 7 imex curation Acetylation is indispensable for p53 activation. Acetylation is indispensable for p53 activation. Cell (0092-8674) 2008 imex curation Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Change to acetylation assay when CVs update Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 7 in vitro In vitro emsa supershift electrophoretic mobility supershift assay emsa supershift predetermined predetermined participant predetermined Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Change to acetylation assay when CVs update Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu Accepted 2008-OCT-29 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 7 imex curation Acetylation is indispensable for p53 activation. Acetylation is indispensable for p53 activation. Cell (0092-8674) 2008 imex curation Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Change to acetylation assay when CVs update Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 7 human-h1299 Homo sapiens lung lymph node carcinoma h1299 Human lung lymph node carcinoma p53 null. These cells have also been reported to secrete the peptide hormone neuromedin B (NMB), but not gastrin releasing peptide (GRP). Also known as NCI H1299 or CRL-5803. http://www.lgcpromochem.com/atcc/ ATCC number: CRL-5803. ATCC number of the cell line is CRL-5803. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Change to acetylation assay when CVs update HA-tagged CREBBP identified using an anti-tag Ab Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu Accepted 2008-OCT-29 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 7 imex curation Acetylation is indispensable for p53 activation. Acetylation is indispensable for p53 activation. Cell (0092-8674) 2008 imex curation Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Change to acetylation assay when CVs update Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 7 human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Change to acetylation assay when CVs update Cells treated with actinomycin D (10 nM) for 0, 4, 8, 16, and 24 hrs. Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu Accepted 2008-OCT-29 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 7 imex curation Acetylation is indispensable for p53 activation. Acetylation is indispensable for p53 activation. Cell (0092-8674) 2008 imex curation Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Change to acetylation assay when CVs update Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 7 in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Change to acetylation assay when CVs update Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu Accepted 2008-OCT-29 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 7 imex curation Acetylation is indispensable for p53 activation. Acetylation is indispensable for p53 activation. Cell (0092-8674) 2008 imex curation Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Change to acetylation assay when CVs update Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 7 in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa predetermined predetermined participant predetermined Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Change to acetylation assay when CVs update Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu Accepted 2008-OCT-29 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 7 imex curation Acetylation is indispensable for p53 activation. Acetylation is indispensable for p53 activation. Cell (0092-8674) 2008 imex curation Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Change to acetylation assay when CVs update Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 7 human-h1299 Homo sapiens lung lymph node carcinoma h1299 Human lung lymph node carcinoma p53 null. These cells have also been reported to secrete the peptide hormone neuromedin B (NMB), but not gastrin releasing peptide (GRP). Also known as NCI H1299 or CRL-5803. http://www.lgcpromochem.com/atcc/ ATCC number: CRL-5803. ATCC number of the cell line is CRL-5803. ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Tang Y., Zhao W., Chen Y., Zhao Y., Gu W. Apoptosis - Interactions involving proteins with a function related to apoptosis Change to acetylation assay when CVs update Source of p53 taken from PMID: 17189186, ep300 from PMID: 17189186 and inferred by sequence length, CBP from PMID: 9194564 and inferred from sequence length. wg8@columbia.edu Accepted 2008-OCT-29 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 7 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 zn363_human_gene Human RCHY1 gene gene gene human Homo sapiens Human mdm2_human_gene Human MDM2 gene gene gene human Homo sapiens Human 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 tp53i3_human_gene Human TP53I3 gene gene gene human Homo sapiens Human cbp_mouse Histone lysine acetyltransferase CREBBP Crebbp Cbp Protein-lysine acetyltransferase CREBBP protein protein mouse Mus musculus Mouse MAENLLDGPPNPKRAKLSSPGFSANDNTDFGSLFDLENDLPDELIPNGELSLLNSGNLVPDAASKHKQLSELLRGGSGSSINPGIGNVSASSPVQQGLGGQAQGQPNSTNMASLGAMGKSPLNQGDSSTPNLPKQAASTSGPTPPASQALNPQAQKQVGLVTSSPATSQTGPGICMNANFNQTHPGLLNSNSGHSLMNQAQQGQAQVMNGSLGAAGRGRGAGMPYPAPAMQGATSSVLAETLTQVSPQMAGHAGLNTAQAGGMTKMGMTGTTSPFGQPFSQTGGQQMGATGVNPQLASKQSMVNSLPAFPTDIKNTSVTTVPNMSQLQTSVGIVPTQAIATGPTADPEKRKLIQQQLVLLLHAHKCQRREQANGEVRACSLPHCRTMKNVLNHMTHCQAGKACQVAHCASSRQIISHWKNCTRHDCPVCLPLKNASDKRNQQTILGSPASGIQNTIGSVGAGQQNATSLSNPNPIDPSSMQRAYAALGLPYMNQPQTQLQPQVPGQQPAQPPAHQQMRTLNALGNNPMSIPAGGITTDQQPPNLISESALPTSLGATNPLMNDGSNSGNIGSLSTIPTAAPPSSTGVRKGWHEHVTQDLRSHLVHKLVQAIFPTPDPAALKDRRMENLVAYAKKVEGDMYESANSRDEYYHLLAEKIYKIQKELEEKRRSRLHKQGILGNQPALPASGAQPPVIPPAQSVRPPNGPLPLPVNRMQVSQGMNSFNPMSLGNVQLPQAPMGPRAASPMNHSVQMNSMASVPGMAISPSRMPQPPNMMGTHANNIMAQAPTQNQFLPQNQFPSSSGAMSVNSVGMGQPAAQAGVSQGQVPGAALPNPLNMLAPQASQLPCPPVTQSPLHPTPPPASTAAGMPSLQHPTAPGMTPPQPAAPTQPSTPVSSGQTPTPTPGSVPSAAQTQSTPTVQAAAQAQVTPQPQTPVQPPSVATPQSSQQQPTPVHTQPPGTPLSQAAASIDNRVPTPSSVTSAETSSQQPGPDVPMLEMKTEVQTDDAEPEPTESKGEPRSEMMEEDLQGSSQVKEETDTTEQKSEPMEVEEKKPEVKVEAKEEEENSSNDTASQSTSPSQPRKKIFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDIVKNPMDLSTIKRKLDTGQYQEPWQYVDDVWLMFNNAWLYNRKTSRVYKFCSKLAEVFEQEIDPVMQSLGYCCGRKYEFSPQTLCCYGKQLCTIPRDAAYYSYQNRYHFCEKCFTEIQGENVTLGDDPSQPQTTISKDQFEKKKNDTLDPEPFVDCKECGRKMHQICVLHYDIIWPSGFVCDNCLKKTGRPRKENKFSAKRLQTTRLGNHLEDRVNKFLRRQNHPEAGEVFVRVVASSDKTVEVKPGMKSRFVDSGEMSESFPYRTKALFAFEEIDGVDVCFFGMHVQNTALIAPHQIQGRVYISYLDSIHFFRPRCLRTAVYHEILIGYLEYVKKLGYVTGHIWACPPSEGDDYIFHCHPPDQKIPKPKRLQEWYKKMLDKAFAERIINDYKDIFKQANEDRLTSAKELPYFEGDFWPNVLEESIKELEQEEEERKKEESTAASETPEGSQGDSKNAKKKNNKKTNKNKSSISRANKKKPSMPNVSNDLSQKLYATMEKHKEVFFVIHLHAGPVISTQPPIVDPDPLLSCDLMDGRDAFLTLARDKHWEFSSLRRSKWSTLCMLVELHTQGQDRFVYTCNECKHHVETRWHCTVCEDYDLCINCYNTKSHTHKMVKWGLGLDDEGSSQGEPQSKSPQESRRLSIQRCIQSLVHACQCRNANCSLPSCQKMKRVVQHTKGCKRKTNGGCPVCKQLIALCCYHAKHCQENKCPVPFCLNIKHKLRQQQIQHRLQQAQLMRRRMATMNTRNVPQQSLPSPTSAPPGTPTQQPSTPQTPQPPAQPQPSPVNMSPAGFPNVARTQPPTIVSAGKPTNQVPAPPPPAQPPPAAVEAARQIEREAQQQQHLYRANINNGMPPGRAGMGTPGSQMTPVGLNVPRPNQVSGPVMSSMPPGQWQQAPIPQQQPMPGMPRPVMSMQAQAAVAGPRMPNVQPPRSISPSALQDLLRTLKSPSSPQQQQQVLNILKSNPQLMAAFIKQRTAKYVANQPGMQPQPGLQSQPGMQPQPGMHQQPSLQNLNAMQAGVPRPGVPPPQPAMGGLNPQGQALNIMNPGHNPNMTNMNPQYREMVRRQLLQHQQQQQQQQQQQQQQQNSASLAGGMAGHSQFQQPQGPGGYAPAMQQQRMQQHLPIQGSSMGQMAAPMGQLGQMGQPGLGADSTPNIQQALQQRILQQQQMKQQIGSPGQPNPMSPQQHMLSGQPQASHLPGQQIATSLSNQVRSPAPVQSPRPQSQPPHSSPSPRIQPQPSPHHVSPQTGSPHPGLAVTMASSMDQGHLGNPEQSAMLPQLNTPNRSALSSELSLVGDTTGDTLEKFVEGL DIP protein P45481 original sequence version: 148 DIP protein P45481 original sequence version: 126 DIP protein P45481 original sequence version: 152 DIP protein P45481 original sequence version: 115 DIP protein P45481 original sequence version: 127 DIP protein P45481 original sequence version: 123 DIP protein P45481 original sequence version: 149 DIP protein P45481 original sequence version: 156 DIP protein P45481 original sequence version: 169 DIP protein P45481 original sequence version: 175 DIP protein P45481 original sequence version: 135 D89AF52B7BD33347 kat5_human Histone acetyltransferase KAT5 KAT5 TIP60 HTATIP Lysine acetyltransferase 5 cPLA(2)-interacting protein Histone acetyltransferase HTATIP 60 kDa Tat-interactive protein protein protein human Homo sapiens Human MAEVGEIIEGCRLPVLRRNQDNEDEWPLAEILSVKDISGRKLFYVHYIDFNKRLDEWVTHERLDLKKIQFPKKEAKTPTKNGLPGSRPGSPEREVPASAQASGKTLPIPVQITLRFNLPKEREAIPGGEPDQPLSSSSCLQPNHRSTKRKVEVVSPATPVPSETAPASVFPQNGAARRAVAAQPGRKRKSNCLGTDEDSQDSSDGIPSAPRMTGSLVSDRSHDDIVTRMKNIECIELGRHRLKPWYFSPYPQELTTLPVLYLCEFCLKYGRSLKCLQRHLTKCDLRHPPGNEIYRKGTISFFEIDGRKNKSYSQNLCLLAKCFLDHKTLYYDTDPFLFYVMTEYDCKGFHIVGYFSKEKESTEDYNVACILTLPPYQRRGYGKLLIEFSYELSKVEGKTGTPEKPLSDLGLLSYRSYWSQTILEILMGLKSESGERPQITINEISEITSIKKEDVISTLQYLNLINYYKGQYILTLSEDIVDGHERAMLKRLLRIDSKCLHFTPKDWSKRGKW DIP protein Q92993 original sequence version: 153 DIP protein Q92993 original sequence version: 155 DIP protein Q92993 original sequence version: 86 DIP protein Q92993 original sequence version: 149 DIP protein Q92993-1 original sequence version: 129 DIP protein Q92993 original sequence version: 144 DIP protein Q92993 original sequence version: 173 63724F5E10B957D5 ep300_human Histone acetyltransferase p300 Histone butyryltransferase p300 Histone crotonyltransferase p300 Protein propionyltransferase p300 EP300 P300 E1A-associated protein p300 Protein 2-hydroxyisobutyryltransferase p300 Protein lactyltransferas p300 protein protein human Homo sapiens Human MAENVVEPGPPSAKRPKLSSPALSASASDGTDFGSLFDLEHDLPDELINSTELGLTNGGDINQLQTSLGMVQDAASKHKQLSELLRSGSSPNLNMGVGGPGQVMASQAQQSSPGLGLINSMVKSPMTQAGLTSPNMGMGTSGPNQGPTQSTGMMNSPVNQPAMGMNTGMNAGMNPGMLAAGNGQGIMPNQVMNGSIGAGRGRQNMQYPNPGMGSAGNLLTEPLQQGSPQMGGQTGLRGPQPLKMGMMNNPNPYGSPYTQNPGQQIGASGLGLQIQTKTVLSNNLSPFAMDKKAVPGGGMPNMGQQPAPQVQQPGLVTPVAQGMGSGAHTADPEKRKLIQQQLVLLLHAHKCQRREQANGEVRQCNLPHCRTMKNVLNHMTHCQSGKSCQVAHCASSRQIISHWKNCTRHDCPVCLPLKNAGDKRNQQPILTGAPVGLGNPSSLGVGQQSAPNLSTVSQIDPSSIERAYAALGLPYQVNQMPTQPQVQAKNQQNQQPGQSPQGMRPMSNMSASPMGVNGGVGVQTPSLLSDSMLHSAINSQNPMMSENASVPSLGPMPTAAQPSTTGIRKQWHEDITQDLRNHLVHKLVQAIFPTPDPAALKDRRMENLVAYARKVEGDMYESANNRAEYYHLLAEKIYKIQKELEEKRRTRLQKQNMLPNAAGMVPVSMNPGPNMGQPQPGMTSNGPLPDPSMIRGSVPNQMMPRITPQSGLNQFGQMSMAQPPIVPRQTPPLQHHGQLAQPGALNPPMGYGPRMQQPSNQGQFLPQTQFPSQGMNVTNIPLAPSSGQAPVSQAQMSSSSCPVNSPIMPPGSQGSHIHCPQLPQPALHQNSPSPVPSRTPTPHHTPPSIGAQQPPATTIPAPVPTPPAMPPGPQSQALHPPPRQTPTPPTTQLPQQVQPSLPAAPSADQPQQQPRSQQSTAASVPTPTAPLLPPQPATPLSQPAVSIEGQVSNPPSTSSTEVNSQAIAEKQPSQEVKMEAKMEVDQPEPADTQPEDISESKVEDCKMESTETEERSTELKTEIKEEEDQPSTSATQSSPAPGQSKKKIFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDIVKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQEIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQGESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAGFVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHASDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPPPNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDYIFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFEGDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLSRGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDLMDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQDRFVYTCNECKHHVETRWHCTVCEDYDLCITCYNTKNHDHKMEKLGLGLDDESNNQQAAATQSPGDSRRLSIQRCIQSLVHACQCRNANCSLPSCQKMKRVVQHTKGCKRKTNGGCPICKQLIALCCYHAKHCQENKCPVPFCLNIKQKLRQQQLQHRLQQAQMLRRRMASMQRTGVVGQQQGLPSPTPATPTTPTGQQPTTPQTPQPTSQPQPTPPNSMPPYLPRTQAAGPVSQGKAAGQVTPPTPPQTAQPPLPGPPPAAVEMAMQIQRAAETQRQMAHVQIFQRPIQHQMPPMTPMAPMGMNPPPMTRGPSGHLEPGMGPTGMQQQPPWSQGGLPQPQQLQSGMPRPAMMSVAQHGQPLNMAPQPGLGQVGISPLKPGTVSQQALQNLLRTLRSPSSPLQQQQVLSILHANPQLLAAFIKQRAAKYANSNPQPIPGQPGMPQGQPGLQPPTMPGQQGVHSNPAMQNMNPMQAGVQRAGLPQQQPQQQLQPPMGGMSPQAQQMNMNHNTMPSQFRDILRRQQMMQQQQQQGAGPGIGPGMANHNQFQQPQGVGYPPQQQQRMQHHMQQMQQGNMGQIGQLPQALGAEAGASLQAYQQRLLQQQMGSPVQPNPMSPQQHMLPNQAQSPHLQGQQIPNSLSNQVRSPQPVPSPRPQSQPPHSSPSPRMQPQPSPHHVSPQTSSPHPGLVAAQANPMEQGHFASPDQNSMLSQLASNPGMANLHGASATDLGLSTDNSDLNSNLSQSTLDIH DIP protein Q09472 original sequence version: 193 DIP protein Q09472 original sequence version: 186 DIP protein Q09472 original sequence version: 107 DIP protein Q09472 original sequence version: 112 DIP protein Q09472 original sequence version: 181 DIP protein Q09472 original sequence version: 183 DIP protein Q09472 original sequence version: 136 DIP protein Q09472 original sequence version: 146 DIP protein Q09472 original sequence version: 165 DIP protein Q09472 original sequence version: 192 DIP protein Q09472 original sequence version: 200 DIP protein Q09472 original sequence version: 206 DIP protein Q09472 original sequence version: 194 DIP protein Q09472 original sequence version: 171 DIP protein Q09472 original sequence version: 142 DIP protein Q09472 original sequence version: 180 8E869E1F174A6FEB cdkn1a_human_gene Human CDKN1A gene gene gene human Homo sapiens Human cdkn1a_human_p53bind Human CDKN1A gene - TP53 binding region dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid human Homo sapiens Human ep300-tp53-1 2 12 enzyme target enzyme target substrate substrate neutral component neutral component 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 undefined mutation mutation mutation certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:13:33 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'lys_arg' 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. 16 enzyme enzyme neutral component neutral component 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 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. acetylation acetylation reaction acetylation In vitro acetylation of p53 by p300/CBP Fig. 1D Acetylation of p53 on lys-164 crebbp-tp53-1 2 14 enzyme enzyme neutral component neutral component 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. 12 enzyme target enzyme target substrate substrate neutral component neutral component 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 undefined mutation mutation mutation certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:21:45 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'lys_arg' P04637:p.Lys164Arg mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K R Wed May 02 16:45:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys164arg' Tue Jun 30 19:07:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys164Arg' 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. acetylation acetylation reaction acetylation In vitro acetylation of p53 by p300/CBP Fig. 1D Acetylation of p53 on lys-164 p53-p21_promoter 7 17 unspecified role unspecified role bait bait promoter region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 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 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. direct interaction direct interaction As shown in Figure 2D, p53/DNA complexes were obtained by incubation of highly purified, recombinant full-length human wild-type p53 with a radiolabeled probe containing the p53-binding site from the p21 promoter. Fig. 2D P04637 transfection p53-mdm2-3 8 12 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey 17 unspecified role unspecified role prey prey association association P04637; Q00987 transfection Mdm2 was only present in the complex when K120, K164, K370, K372, K373, K381, K382, and K386 were all mutated to alanine, preventing acetylation suggesting that TP53 acetylation represses recruitment of MDM2 to the CDKN1A promoter. Mdm2 recruitment at the p21 promoter was strongly repressed upon treatment with the DNA-damaging reagent actinomycin D which correlated with the acetylation levels of wild-type p53 To elucidate the mechanism of p53 acetylation in p21 activation in cells, we performed chromatin immunoprecipitation (ChIP) assays with cells expressing either wild-type or acetylation-defective p53 proteins. Fig. 4A, 4C, 6D p53-crebbp-8 8 14 unspecified role unspecified role prey prey 15 unspecified role unspecified role prey prey 17 unspecified role unspecified role prey prey 12 unspecified role unspecified role bait bait association association P04637; Q92993, P45481 transfection nterestingly, coexpression of the CBP and Tip60 acetylases with wild-type p53, which led to increasing levels of p53 acetylation (lane 4, Figure 4B) Fig. 4A, 4C p53-crebbp-9 8 9 unspecified role unspecified role prey prey 12 unspecified role unspecified role bait bait 14 unspecified role unspecified role prey prey 15 unspecified role unspecified role prey prey 13 unspecified role unspecified role prey prey association association P04637; Q92993, P45481; Q00987 transfection The occupancy of p53, Mdm2, Mdmx, CBP, and Tip60 of the p21, Pig3, and Mdm2 promoters was detected by PCR-agarose gel electrophoresis. Fig. 4C MDM4 (MDMX) was also observed to bid in this promoter complex but the source could not be identified. p53-crebbp-10 8 9 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 12 unspecified role unspecified role bait bait 14 unspecified role unspecified role prey prey association association P04637; P45481 transfection The occupancy of p53, Mdm2, Mdmx, CBP, and Tip60 of the p21, Pig3, and Mdm2 promoters was detected by PCR-agarose gel electrophoresis. Fig. 4C, Supp. Fig. 9 Recruitment of MDM2 to the MDM2 promoteris fairly weak and requires further cycles of PCR to visualise. p53-p21_p53bind 7 12 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 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. 9 unspecified role unspecified role prey prey region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 18 unspecified role unspecified role bait bait association association An EMSA was then used to examine whether Mdm2 can form a protein complex with DNAbound p53. Fig. 5B DNA is 32P-radiolabelled Q00987; P04637 transfection Use of the lys to arg mutant protein demonstracted that MDM2 bound preferably to the unaccetylated TP53 protein. In contrast, acetylated p53 bound DNA more strongly. p53-p21_p53bind-1 3 12 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 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. 9 unspecified role unspecified role prey prey region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 18 unspecified role unspecified role bait bait association association this complex was either supershifted by the p53-specific antibody or blocked by the Mdm2-specific antibody Fig. 5B DNA is 32P-radiolabelled Q00987; P04637 transfection The complex was supershifted by a p53-specific antibody or blocked by an Mdm2-specific antibody mdm2-tp53-2 1 9 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 12 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined dna binding region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction As expected, 35S-labeled in vitro-translated Mdm2 bound the recombinant protein GST-p53 (aa 1�73) (lane 2, Figure 5D) but not GST (lane 5, Figure 5D). Surprisingly, both the core domain and the C terminus of p53 showed similar binding affinities for Mdm2 (lanes 3 and 4, Figure 5D), Fig. 5D mdm2-tp53-3 6 12 unspecified role unspecified role prey prey 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. 9 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction More importantly, to examine the effect of acetylation in this interaction, we performed similar GST-pulldown assays with either purified unacetylated p53 or acetylated p53 proteins. Fig. 5E Acetylation of TP53 inhibited this interaction An interaction between MDM4 and TP53 was also demonstrated but the source of MDM4 could not be traced. p53-mdm2-4 5 12 unspecified role unspecified role bait bait 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 17 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey association association Increased TP53 acetylation due to DNA damage represses recruitment of MDM2 to the CDKN1A promoter. As expected, immunoprecipitation with the p53 antibody showed that an increasing amount of p53 associated with the p21 promoter (Figure 6B); conversely, although the levels of Mdm2 were elevated (Figure 6A), the recruitment of Mdm2 to the p21 promoter by p53 was severely inhibited when p53 was acetylated (Figure 6B). Fig. 6B tp53-crebbp 4 14 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 12 unspecified role unspecified role bait bait 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 physical association physical association Moreover, both forms of p53 also bound equally to the transcriptional coactivators/acetylases CBP and Tip60 (Figures S3A and S3B), Supp. Fig. 3A tp53-tip60-1 4 12 unspecified role unspecified role bait bait 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 15 unspecified role unspecified role prey prey physical association physical association Moreover, both forms of p53 also bound equally to the transcriptional coactivators/acetylases CBP and Tip60 (Figures S3A and S3B), Supp. Fig. 3A P04637; Q92993 transfection mdm2-tp53-4 1 9 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 12 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction In vitro binding of p53 or p53-8KR to Mdm2 Supp. Fig. 4 An interaction between MDM4 and TP53 was also demonstrated but the source of MDM4 could not be traced. tp53-rchy1_gene 8 14 unspecified role unspecified role prey prey 12 unspecified role unspecified role bait bait 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 10 unspecified role unspecified role prey prey association association P04637; P45481 transfection Supp. Fig. 10B Pirh2 as another p53 target (Leng et al., 2003), whose induction by p53 is independent of the acetylation status (Figure S10).