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/ zou-2014-1 Brd4 maintains constitutively active NF-κB in cancer cells by binding to acetylated RelA. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Zou Z., Huang B., Wu X., Zhang H., Qi J., Bradner J., Nair S., Chen L.F. IBD - Inflammatory bowel disease Expression and purification of BRD4 bromodomains (BD1 and BD2) is described in PMID:19103749. lfchen@life.illinois.edu Oncogene (0950-9232) 2014 Only protein-protein interactions imex curation zou-2014-2 Brd4 maintains constitutively active NF-κB in cancer cells by binding to acetylated RelA. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Zou Z., Huang B., Wu X., Zhang H., Qi J., Bradner J., Nair S., Chen L.F. IBD - Inflammatory bowel disease Expression and purification of BRD4 bromodomains (BD1 and BD2) as well as recombinant RelA are described in PMID:19103749. Recombinant RelA was expressed in insect ceels, but the specific species was not given by the authors, so this is not registered in the "expressed in" field. lfchen@life.illinois.edu Oncogene (0950-9232) 2014 Only protein-protein interactions imex curation zou-2014-3 Brd4 maintains constitutively active NF-κB in cancer cells by binding to acetylated RelA. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Zou Z., Huang B., Wu X., Zhang H., Qi J., Bradner J., Nair S., Chen L.F. IBD - Inflammatory bowel disease Expression and purification of BRD4 bromodomains (BD1 and BD2) as well as the synthetic RelA construct are described in PMID:19103749. lfchen@life.illinois.edu Oncogene (0950-9232) 2014 Only protein-protein interactions imex curation zou-2014-4 Brd4 maintains constitutively active NF-κB in cancer cells by binding to acetylated RelA. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Zou Z., Huang B., Wu X., Zhang H., Qi J., Bradner J., Nair S., Chen L.F. IBD - Inflammatory bowel disease The experiment and protein constructs are described in PMID:19103749 and PMID:16135789. lfchen@life.illinois.edu Oncogene (0950-9232) 2014 Only protein-protein interactions imex curation zou-2014-5 Brd4 maintains constitutively active NF-κB in cancer cells by binding to acetylated RelA. human-a_549 Homo sapiens Caucasian lung carcinoma cell line a_549 Human Caucasian lung carcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zou Z., Huang B., Wu X., Zhang H., Qi J., Bradner J., Nair S., Chen L.F. IBD - Inflammatory bowel disease The experiment was performed using nuclear extracts of A549 cells. lfchen@life.illinois.edu Oncogene (0950-9232) 2014 Only protein-protein interactions imex curation tf65_human Transcription factor p65 Nuclear factor NF-kappa-B p65 subunit RELA NFKB3 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 protein protein human Homo sapiens Human MDELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGSIPGERSTDTTKTHPTIKINGYTGPGTVRISLVTKDPPHRPHPHELVGKDCRDGFYEAELCPDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNPFQVPIEEQRGDYDLNAVRLCFQVTVRDPSGRPLRLPPVLSHPIFDNRAPNTAELKICRVNRNSGSCLGGDEIFLLCDKVQKEDIEVYFTGPGWEARGSFSQADVHRQVAIVFRTPPYADPSLQAPVRVSMQLRRPSDRELSEPMEFQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISS brd4_human Bromodomain-containing protein 4 Protein HUNK1 BRD4 HUNK1 protein protein human Homo sapiens Human MSAESGPGTRLRNLPVMGDGLETSQMSTTQAQAQPQPANAASTNPPPPETSNPNKPKRQTNQLQYLLRVVLKTLWKHQFAWPFQQPVDAVKLNLPDYYKIIKTPMDMGTIKKRLENNYYWNAQECIQDFNTMFTNCYIYNKPGDDIVLMAEALEKLFLQKINELPTEETEIMIVQAKGRGRGRKETGTAKPGVSTVPNTTQASTPPQTQTPQPNPPPVQATPHPFPAVTPDLIVQTPVMTVVPPQPLQTPPPVPPQPQPPPAPAPQPVQSHPPIIAATPQPVKTKKGVKRKADTTTPTTIDPIHEPPSLPPEPKTTKLGQRRESSRPVKPPKKDVPDSQQHPAPEKSSKVSEQLKCCSGILKEMFAKKHAAYAWPFYKPVDVEALGLHDYCDIIKHPMDMSTIKSKLEAREYRDAQEFGADVRLMFSNCYKYNPPDHEVVAMARKLQDVFEMRFAKMPDEPEEPVVAVSSPAVPPPTKVVAPPSSSDSSSDSSSDSDSSTDDSEEERAQRLAELQEQLKAVHEQLAALSQPQQNKPKKKEKDKKEKKKEKHKRKEEVEENKKSKAKEPPPKKTKKNNSSNSNVSKKEPAPMKSKPPPTYESEEEDKCKPMSYEEKRQLSLDINKLPGEKLGRVVHIIQSREPSLKNSNPDEIEIDFETLKPSTLRELERYVTSCLRKKRKPQAEKVDVIAGSSKMKGFSSSESESSSESSSSDSEDSETEMAPKSKKKGHPGREQKKHHHHHHQQMQQAPAPVPQQPPPPPQQPPPPPPPQQQQQPPPPPPPPSMPQQAAPAMKSSPPPFIATQVPVLEPQLPGSVFDPIGHFTQPILHLPQPELPPHLPQPPEHSTPPHLNQHAVVSPPALHNALPQQPSRPSNRAAALPPKPARPPAVSPALTQTPLLPQPPMAQPPQVLLEDEEPPAPPLTSMQMQLYLQQLQKVQPPTPLLPSVKVQSQPPPPLPPPPHPSVQQQLQQQPPPPPPPQPQPPPQQQHQPPPRPVHLQPMQFSTHIQQPPPPQGQQPPHPPPGQQPPPPQPAKPQQVIQHHHSPRHHKSDPYSTGHLREAPSPLMIHSPQMSQFQSLTHQSPPQQNVQPKKQELRAASVVQPQPLVVVKEEKIHSPIIRSEPFSPSLRPEPPKHPESIKAPVHLPQRPEMKPVDVGRPVIRPPEQNAPPPGAPDKDKQKQEPKTPVAPKKDLKIKNMGSWASLVQKHPTTPSSTAKSSSDSFEQFRRAAREKEEREKALKAQAEHAEKEKERLRQERMRSREDEDALEQARRAHEEARRRQEQQQQQRQEQQQQQQQQAAAVAAAATPQAQSSQPQSMLDQQRELARKREQERRRREAMAATIDMNFQSDLLSIFEENLF brd4-rela 1571246 n/a RelA 1571251 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false possible acetyllys-310 n6aclys N6-acetyl-L-lysine (2S)-6-acetamido-2-aminohexanoic acid 6-acetylamino-2-aminohexanoic acid 6-acetylazanyl-2-aminohexanoic acid MOD_RES N6-acetyllysine N(zeta)-acetyllysine N6-acetyl-L-lysine N6-acetylated L-lysine epsilon-acetyllysine Acetyl Acetylation N6AcLys acetylk predetermined featur predetermined feature predetermined featur certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 2.0 n/a 1571252 unspecified role unspecified role neutral component neutral component bromodomain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) Stoichiometry: 2.0 1571258 1571255 direct interaction direct interaction false false false Suppl. Table 1 1.50A working 19.8%, free 22.2% O60885 increases activity of Q04206 O60885 decreases degradation at protein level of Q04206 brd4-rela-1 1571246 n/a 1571252 unspecified role unspecified role neutral component neutral component bromodomain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) Stoichiometry: 1.0 n/a RelA 1571251 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false possible acetyllys-310 n6aclys N6-acetyl-L-lysine (2S)-6-acetamido-2-aminohexanoic acid 6-acetylamino-2-aminohexanoic acid 6-acetylazanyl-2-aminohexanoic acid MOD_RES N6-acetyllysine N(zeta)-acetyllysine N6-acetyl-L-lysine N6-acetylated L-lysine epsilon-acetyllysine Acetyl Acetylation N6AcLys acetylk predetermined featur predetermined feature predetermined featur certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 1571263 1571261 direct interaction direct interaction false false false Suppl. Table 1 2.00A working 20.8%, free 25.8% brd4-rela-3 1571247 n/a 1571252 unspecified role unspecified role bait bait bromodomain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false O60885:p.Tyr139_Asn140delinsAlaAla mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false O60885:p.Tyr432_Asn433delinsAlaAla mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false O60885:p.[Tyr139_Asn140delinsAlaAla;Tyr432_Asn433delinsAlaAla] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) n/a RelA 1571251 unspecified role unspecified role prey prey possible acetyllys-310 n6aclys N6-acetyl-L-lysine (2S)-6-acetamido-2-aminohexanoic acid 6-acetylamino-2-aminohexanoic acid 6-acetylazanyl-2-aminohexanoic acid MOD_RES N6-acetyllysine N(zeta)-acetyllysine N6-acetyl-L-lysine N6-acetylated L-lysine epsilon-acetyllysine Acetyl Acetylation N6AcLys acetylk predetermined featur predetermined feature predetermined featur certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig. 1c rela-brd4 1571248 n/a 1571252 unspecified role unspecified role prey prey bromodomain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false ecobb Escherichia coli (strain B / BL21) n/a RelA 1571251 unspecified role unspecified role bait bait possible acetyllys-310 n6aclys N6-acetyl-L-lysine (2S)-6-acetamido-2-aminohexanoic acid 6-acetylamino-2-aminohexanoic acid 6-acetylazanyl-2-aminohexanoic acid MOD_RES N6-acetyllysine N(zeta)-acetyllysine N6-acetyl-L-lysine N6-acetylated L-lysine epsilon-acetyllysine Acetyl Acetylation N6AcLys acetylk 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 region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) 1571280 1571276 direct interaction direct interaction false false false Fig. 2a This interaction was inhibited by addition of JQ1 (2 micromolar), a member of the BET (bromodomain extra terminal) protein inhibitor family. brd4-rela-4 1571249 n/a 1571252 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 false n/a RelA 1571251 unspecified role unspecified role prey prey region t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false possible acetyllys-310 n6aclys N6-acetyl-L-lysine (2S)-6-acetamido-2-aminohexanoic acid 6-acetylamino-2-aminohexanoic acid 6-acetylazanyl-2-aminohexanoic acid MOD_RES N6-acetyllysine N(zeta)-acetyllysine N6-acetyl-L-lysine N6-acetylated L-lysine epsilon-acetyllysine Acetyl Acetylation N6AcLys acetylk western blot western blot Immuno blot Immuno blot certain certain sequence position certain certain certain sequence position certain false acetylation at lys310 physical association physical association false false false Fig. 2b This interaction was inhibited by addition of JQ1 (10 and 20 micromolar), a member of the BET (bromodomain extra terminal) protein inhibitor family. This interaction was enhanced by co-transfection of cells with p300. rela-brd4-1 1571250 n/a 1571251 unspecified role unspecified role bait bait n/a 1571252 unspecified role unspecified role prey prey physical association physical association false false false Fig. 2c This interaction was inhibited by treatment of cells with JQ1 (5 micromolar for 4 hours), a member of the BET (bromodomain extra terminal) protein inhibitor family.