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/ Mutations in host cell factor 1 separate its role in cell proliferation from recruitment of VP16 and LZIP. Mutations in host cell factor 1 separate its role in cell proliferation from recruitment of VP16 and LZIP. Mol. Cell. Biol. (0270-7306) 2000 Mahajan SS., Wilson AC. Fig 6 wilsoa02@popmail.med.nyu.edu in vitro In vitro coip coimmunoprecipitation co-immunoprecipitation CoIp immunoprecipitation Co-IP co-immunoprecipitation CoIp immunoprecipitation Co-IP coip predetermined predetermined participant predetermined Mahajan SS., Wilson AC. Fig 6 wilsoa02@popmail.med.nyu.edu Mol. Cell. Biol. (0270-7306) 2000 Mutations in host cell factor 1 separate its role in cell proliferation from recruitment of VP16 and LZIP. Mutations in host cell factor 1 separate its role in cell proliferation from recruitment of VP16 and LZIP. Mol. Cell. Biol. (0270-7306) 2000 Mahajan SS., Wilson AC. Fig 6 wilsoa02@popmail.med.nyu.edu 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. biophysical biophysical predetermined predetermined participant predetermined Mahajan SS., Wilson AC. Revisit when on-hybrid is in place A cell-based recruitment assay was used to compare the sensitivities of vp16 and creb3 binding to each of the hcfc1-vic (vp16-induced complex) domain mutants. This 'one-hybrid' recruitment assay was used because creb3 is expressed at very low levels in transfected cells, making it difficult to quantitate association with cotransfected hcfc1 mutants by coimmunoprecipitation. The Hcfc1 beta-propeller domain was expressed in 293T cells as a GAL4 fusion together with either vp16 (AA 5-490) or the N-terminus of creb3 (AA 1-154). Recruitment was measured in terms of activation of a Gal4-responsive luciferase reporter gene and the amounts of each expression plasmid chosen to give a linear reponse. Fig 6 wilsoa02@popmail.med.nyu.edu Mol. Cell. Biol. (0270-7306) 2000 creb3_human Cyclic AMP-responsive element-binding protein 3 Luman Transcription factor LZIP-alpha CREB3 LZIP Leucine zipper protein protein protein human Homo sapiens Human MELELDAGDQDLLAFLLEESGDLGTAPDEAVRAPLDWALPLSEVPSDWEVDDLLCSLLSPPASLNILSSSNPCLVHHDHTYSLPRETVSMDLESESCRKEGTQMTPQHMEELAEQEIARLVLTDEEKSLLEKEGLILPETLPLTKTEEQILKRVRRKIRNKRSAQESRRKKKVYVGGLESRVLKYTAQNMELQNKVQLLEEQNLSLLDQLRKLQAMVIEISNKTSSSSTCILVLLVSFCLLLVPAMYSSDTRGSLPAEHGVLSRQLRALPSEDPYQLELPALQSEVPKDSTHQWLDGSDCVLQAPGNTSCLLHYMPQAPSAEPPLEWPFPDLFSEPLCRGPILPLQANLTRKGGWLPTGSPSVILQDRYSG DIP protein O43889 original sequence version: 133 82152E496B924EEC po2f1_human POU domain, class 2, transcription factor 1 Octamer-binding transcription factor 1 POU2F1 OCT1 OTF1 Octamer-binding protein 1 NF-A1 protein protein human Homo sapiens Human MNNPSETSKPSMESGDGNTGTQTNGLDFQKQPVPVGGAISTAQAQAFLGHLHQVQLAGTSLQAAAQSLNVQSKSNEESGDSQQPSQPSQQPSVQAAIPQTQLMLAGGQITGLTLTPAQQQLLLQQAQAQAQLLAAAVQQHSASQQHSAAGATISASAATPMTQIPLSQPIQIAQDLQQLQQLQQQNLNLQQFVLVHPTTNLQPAQFIISQTPQGQQGLLQAQNLLTQLPQQSQANLLQSQPSITLTSQPATPTRTIAATPIQTLPQSQSTPKRIDTPSLEEPSDLEELEQFAKTFKQRRIKLGFTQGDVGLAMGKLYGNDFSQTTISRFEALNLSFKNMCKLKPLLEKWLNDAENLSSDSSLSSPSALNSPGIEGLSRRRKKRTSIETNIRVALEKSFLENQKPTSEEITMIADQLNMEKEVIRVWFCNRRQKEKRINPPSSGGTSSSPIKAIFPSPTSLVATTPSLVTSSAATTLTVSPVLPLTSAAVTNLSVTGTSDTTSNNTATVISTAPPASSAVTSPSLSPSPSASASTSEASSASETSTTQTTSTPLSSPLGTSQVMVTASGLQTAAAAALQGAAQLPANASLAAMAAAAGLNPSLMAPSQFAAGGALLSLNPGTLSGALSPALMSNSTLATIQALASGGSLPITSLDATGNLVFANAGGAPNIVTAPLFLNPQNLSLLTSNPVSLVSAAAASAGNSAPVASLHATSTSAESIQNSLFTVASASGAASTTTTASKAQ FAB27FFC79CF2276 hcfc1_human Host cell factor 1 HCFC1 HCF1 C1 factor VP16 accessory protein VCAF CFF HFC1 protein protein human Homo sapiens Human MASAVSPANLPAVLLQPRWKRVVGWSGPVPRPRHGHRAVAIKELIVVFGGGNEGIVDELHVYNTATNQWFIPAVRGDIPPGCAAYGFVCDGTRLLVFGGMVEYGKYSNDLYELQASRWEWKRLKAKTPKNGPPPCPRLGHSFSLVGNKCYLFGGLANDSEDPKNNIPRYLNDLYILELRPGSGVVAWDIPITYGVLPPPRESHTAVVYTEKDNKKSKLVIYGGMSGCRLGDLWTLDIDTLTWNKPSLSGVAPLPRSLHSATTIGNKMYVFGGWVPLVMDDVKVATHEKEWKCTNTLACLNLDTMAWETILMDTLEDNIPRARAGHCAVAINTRLYIWSGRDGYRKAWNNQVCCKDLWYLETEKPPPPARVQLVRANTNSLEVSWGAVATADSYLLQLQKYDIPATAATATSPTPNPVPSVPANPPKSPAPAAAAPAVQPLTQVGITLLPQAAPAPPTTTTIQVLPTVPGSSISVPTAARTQGVPAVLKVTGPQATTGTPLVTMRPASQAGKAPVTVTSLPAGVRMVVPTQSAQGTVIGSSPQMSGMAALAAAAAATQKIPPSSAPTVLSVPAGTTIVKTMAVTPGTTTLPATVKVASSPVMVSNPATRMLKTAAAQVGTSVSSATNTSTRPIITVHKSGTVTVAQQAQVVTTVVGGVTKTITLVKSPISVPGGSALISNLGKVMSVVQTKPVQTSAVTGQASTGPVTQIIQTKGPLPAGTILKLVTSADGKPTTIITTTQASGAGTKPTILGISSVSPSTTKPGTTTIIKTIPMSAIITQAGATGVTSSPGIKSPITIITTKVMTSGTGAPAKIITAVPKIATGHGQQGVTQVVLKGAPGQPGTILRTVPMGGVRLVTPVTVSAVKPAVTTLVVKGTTGVTTLGTVTGTVSTSLAGAGGHSTSASLATPITTLGTIATLSSQVINPTAITVSAAQTTLTAAGGLTTPTITMQPVSQPTQVTLITAPSGVEAQPVHDLPVSILASPTTEQPTATVTIADSGQGDVQPGTVTLVCSNPPCETHETGTTNTATTTVVANLGGHPQPTQVQFVCDRQEAAASLVTSTVGQQNGSVVRVCSNPPCETHETGTTNTATTATSNMAGQHGCSNPPCETHETGTTNTATTAMSSVGANHQRDARRACAAGTPAVIRISVATGALEAAQGSKSQCQTRQTSATSTTMTVMATGAPCSAGPLLGPSMAREPGGRSPAFVQLAPLSSKVRLSSPSIKDLPAGRHSHAVSTAAMTRSSVGAGEPRMAPVCESLQGGSPSTTVTVTALEALLCPSATVTQVCSNPPCETHETGTTNTATTSNAGSAQRVCSNPPCETHETGTTHTATTATSNGGTGQPEGGQQPPAGRPCETHQTTSTGTTMSVSVGALLPDATSSHRTVESGLEVAAAPSVTPQAGTALLAPFPTQRVCSNPPCETHETGTTHTATTVTSNMSSNQDPPPAASDQGEVESTQGDSVNITSSSAITTTVSSTLTRAVTTVTQSTPVPGPSVPPPEELQVSPGPRQQLPPRQLLQSASTALMGESAEVLSASQTPELPAAVDLSSTGEPSSGQESAGSAVVATVVVQPPPPTQSEVDQLSLPQELMAEAQAGTTTLMVTGLTPEELAVTAAAEAAAQAAATEEAQALAIQAVLQAAQQAVMGTGEPMDTSEAAATVTQAELGHLSAEGQEGQATTIPIVLTQQELAALVQQQQLQEAQAQQQHHHLPTEALAPADSLNDPAIESNCLNELAGTVPSTVALLPSTATESLAPSNTFVAPQPVVVASPAKLQAAATLTEVANGIESLGVKPDLPPPPSKAPMKKENQWFDVGVIKGTNVMVTHYFLPPDDAVPSDDDLGTVPDYNQLKKQELQPGTAYKFRVAGINACGRGPFSEISAFKTCLPGFPGAPCAIKISKSPDGAHLTWEPPSVTSGKIIEYSVYLAIQSSQAGGELKSSTPAQLAFMRVYCGPSPSCLVQSSSLSNAHIDYTTKPAIIFRIAARNEKGYGPATQVRWLQETSKDSSGTKPANKRPMSSPEMKSAPKKSKADGQ 0B0C581E2454631E hcfc1-pou2f1-1 1 4 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 5 unspecified role unspecified role bait bait P51610:p.Arg344_Lys345delinsAlaAla mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain RK AA Wed May 02 15:38:16 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg_lys344-345ala_ala' Tue Jun 30 16:13:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg344_Lys345delinsAlaAla' P51610:p.Pro134Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain P S Wed May 02 15:24:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro134ser' Tue Jun 30 18:19:53 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro134Ser' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P51610:p.Glu289_Lys291delinsAlaAlaAla mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain EWK AAA Wed May 02 15:16:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu_trp_lys289-291ala_ala_ala' Tue Jun 30 18:54:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu289_Lys291delinsAlaAlaAla' n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag western blot western blot Immuno blot Immuno blot n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 physical association physical association These results indicate that hcfc1 and pou2f1 can interact weakly in the absence of vp16, and that the residues (AA 344-345)RK within hcfc1 are required for this interaction. hcfc1-creb3-1 2 3 unspecified role unspecified role prey prey n-terminus binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 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. 5 unspecified role unspecified role bait bait P51610:p.Pro79Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain P S Wed May 02 19:34:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro79ser' Tue Jun 30 20:00:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro79Ser' gal4bd tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P51610:p.Pro30Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain P S Wed May 02 19:23:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro30ser' Tue Jun 30 19:02:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro30Ser' P51610:p.Pro134Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain P S Wed May 02 17:33:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro134ser' Tue Jun 30 16:00:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro134Ser' P51610:p.Pro197Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain P S Wed May 02 19:30:59 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro197ser' Tue Jun 30 19:35:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro197Ser' P51610:p.Pro319Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain P S Wed May 02 20:21:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro319ser' Tue Jun 30 15:47:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro319Ser' P51610:p.Pro252Ser mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain P S Wed May 02 15:05:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro252ser' Tue Jun 30 15:57:35 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro252Ser' P51610:p.Cys82Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C D Wed May 02 16:29:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys82asp' Tue Jun 30 18:44:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys82Asp' P51610:p.Arg228Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R D Wed May 02 14:27:06 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg228asp' Tue Jun 30 16:12:57 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg228Asp' P51610:p.Glu289_Lys291delinsAlaAlaAla mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain EWK AAA Wed May 02 19:56:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu_trp_lys289-291ala_ala_ala' Tue Jun 30 18:11:51 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu289_Lys291delinsAlaAlaAla' P51610:p.Ser338Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain S A Wed May 02 14:50:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser338ala' Tue Jun 30 19:11:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser338Ala' n-terminus binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 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. physical association physical association Although vp16 and creb3 use a related sequence motif (the hcfc1 binding motif - HBM) to associate with hcfc1, there were striking differences in their sensitivities to individual point mutations in the hcfc1 beta-propeller. P79S, C82D and EWK289A3 disrupted the recruitment of creb3 more than vp16, to less than 10% of the wild-type level. There was reduced recruitment to less than 10% of wild-type for both vp16 and creb3 which occured with P30S, P134S, P197S, P319S, R228D. S338A caused reduced recruitment to less than 50% and the remaining mutations (P252S, K105D and R344A) had a mild effect on reducing recruitment which did not fall below 70% of wild type.