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/ Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cell (0092-8674) 2009 imex curation Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com 01-Sep-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 19 in vitro In vitro enzymatic study enzymatic study predetermined predetermined participant predetermined Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com Accepted 2009-SEP-18 by JYOTI. 01-Sep-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 19 imex curation Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cell (0092-8674) 2009 imex curation Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com 01-Sep-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 19 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 western blot western blot Immuno blot Immuno blot Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis origin of CUL3 confirmed by author. aa@gene.com Accepted 2009-SEP-18 by JYOTI. 01-Sep-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 19 imex curation Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cell (0092-8674) 2009 imex curation Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com 01-Sep-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 19 human-h2122 Human lung carcinoma cell line h2122 Lung carcinoma cell line H2122 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com Accepted 2009-SEP-18 by JYOTI. 01-Sep-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 19 imex curation Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cell (0092-8674) 2009 imex curation Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com 01-Sep-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 19 human-h460 Homo sapiens non-small-cell lung cancer cell line H460 h460 human non-small-cell lung cancer cell line H460 confocal microscopy confocal microscopy immunostaining immunostaining Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com Accepted 2009-SEP-18 by JYOTI. 01-Sep-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 19 imex curation Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cell (0092-8674) 2009 imex curation Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com 01-Sep-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 19 human-h460 Homo sapiens non-small-cell lung cancer cell line H460 h460 human non-small-cell lung cancer cell line H460 anti bait coip anti bait coimmunoprecipitation anti bait coip de novo protein sequence de novo protein sequencing by mass spectrometry de novo protein sequence Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com Accepted 2009-SEP-18 by JYOTI. 01-Sep-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 19 imex curation Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cell (0092-8674) 2009 imex curation Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com 01-Sep-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 19 human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference confocal microscopy confocal microscopy immunostaining immunostaining Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com Accepted 2009-SEP-18 by JYOTI. 01-Sep-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 19 imex curation Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cell (0092-8674) 2009 imex curation Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com 01-Sep-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 19 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 Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com Accepted 2009-SEP-18 by JYOTI. 01-Sep-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 19 imex curation Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cell (0092-8674) 2009 imex curation Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com 01-Sep-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 19 human-h460 Homo sapiens non-small-cell lung cancer cell line H460 h460 human non-small-cell lung cancer cell line H460 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com Accepted 2009-SEP-18 by JYOTI. 01-Sep-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 19 imex curation Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cell (0092-8674) 2009 imex curation Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com 01-Sep-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 19 human-h460 Homo sapiens non-small-cell lung cancer cell line H460 h460 human non-small-cell lung cancer cell line H460 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Jin Z., Li Y., Pitti R., Lawrence D., Pham VC., Lill JR., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis aa@gene.com Accepted 2009-SEP-18 by JYOTI. 01-Sep-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 19 imex curation ub2l3_human Ubiquitin-conjugating enzyme E2 L3 Ubiquitin-protein ligase L3 Ubiquitin carrier protein L3 UbcH7 Ubiquitin-conjugating enzyme E2-F1 L-UBC UBE2L3 UBCE7 UBCH7 E2 ubiquitin-conjugating enzyme L3 protein protein human Homo sapiens Human MAASRRLMKELEEIRKCGMKNFRNIQVDEANLLTWQGLIVPDNPPYDKGAFRIEINFPAEYPFKPPKITFKTKIYHPNIDEKGQVCLPVISAENWKPATKTDQVIQSLIALVNDPQPEHPLRADLAEEYSKDRKKFCKNAEEFTKKYGEKRPVD F5A30243BE3C9985 casp8_human Caspase-8 CASP8 MCH5 ICE-like apoptotic protease 5 MORT1-associated ced-3 homolog FADD-homologous ICE/ced-3-like protease FADD-like ICE Apoptotic cysteine protease Apoptotic protease Mch-5 CAP4 protein protein human Homo sapiens Human MDFSRNLYDIGEQLDSEDLASLKFLSLDYIPQRKQEPIKDALMLFQRLQEKRMLEESNLSFLKELLFRINRLDLLITYLNTRKEEMERELQTPGRAQISAYRVMLYQISEEVSRSELRSFKFLLQEEISKCKLDDDMNLLDIFIEMEKRVILGEGKLDILKRVCAQINKSLLKIINDYEEFSKERSSSLEGSPDEFSNGEELCGVMTISDSPREQDSESQTLDKVYQMKSKPRGYCLIINNHNFAKAREKVPKLHSIRDRNGTHLDAGALTTTFEELHFEIKPHDDCTVEQIYEILKIYQLMDHSNMDCFICCILSHGDKGIIYGTDGQEAPIYELTSQFTGLKCPSLAGKPKVFFIQACQGDNYQKGIPVETDSEEQPYLEMDLSSPQTRYIPDEADFLLGMATVNNCVSYRNPAEGTWYIQSLCQSLRERCPRGDDILTILTEVNYEVSNKDDKKNMGKQMPQPTFTLRKKLVFPSD DIP protein Q14790 original sequence version: 184 DIP protein Q14790 original sequence version: 135 DIP protein Q14790 original sequence version: 136 DIP protein Q14790 original sequence version: 147 DIP protein Q14790 original sequence version: 146 DIP protein Q14790 original sequence version: 170 DIP protein Q14790 original sequence version: 198 DIP protein Q14790-1 original sequence version: 197 DIP protein Q14790 original sequence version: 158 7A5FEAA6B39B582F sqstm_human Sequestosome-1 SQSTM1 ORCA OSIL Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa Ubiquitin-binding protein p62 EBI3-associated protein of 60 kDa protein protein human Homo sapiens Human MASLTVKAYLLGKEDAAREIRRFSFCCSPEPEAEAEAAAGPGPCERLLSRVAALFPALRPGGFQAHYRDEDGDLVAFSSDEELTMAMSYVKDDIFRIYIKEKKECRRDHRPPCAQEAPRNMVHPNVICDGCNGPVVGTRYKCSVCPDYDLCSVCEGKGLHRGHTKLAFPSPFGHLSEGFSHSRWLRKVKHGHFGWPGWEMGPPGNWSPRPPRAGEARPGPTAESASGPSEDPSVNFLKNVGESVAAALSPLGIEVDIDVEHGGKRSRLTPVSPESSSTEEKSSSQPSSCCSDPSKPGGNVEGATQSLAEQMRKIALESEGRPEEQMESDNCSGGDDDWTHLSSKEVDPSTGELQSLQMPESEGPSSLDPSQEGPTGLKEAALYPHLPPEADPRLIESLSQMLSMGFSDEGGWLTRLLQTKNYDIGAALDTIQYSKHPPPL DIP protein Q13501 original sequence version: 139 DIP protein Q13501 original sequence version: 142 DIP protein Q13501 original sequence version: 163 DIP protein Q13501 original sequence version: 146 DIP protein Q13501 original sequence version: 162 DIP protein Q13501 original sequence version: 155 DIP protein Q13501 original sequence version: 140 462D94C171F337CD caspa_human Caspase-10 ICE-like apoptotic protease 4 Apoptotic protease Mch-4 FAS-associated death domain protein interleukin-1B-converting enzyme 2 CASP10 MCH4 protein protein human Homo sapiens Human MKSQGQHWYSSSDKNCKVSFREKLLIIDSNLGVQDVENLKFLCIGLVPNKKLEKSSSASDVFEHLLAEDLLSEEDPFFLAELLYIIRQKKLLQHLNCTKEEVERLLPTRQRVSLFRNLLYELSEGIDSENLKDMIFLLKDSLPKTEMTSLSFLAFLEKQGKIDEDNLTCLEDLCKTVVPKLLRNIEKYKREKAIQIVTPPVDKEAESYQGEEELVSQTDVKTFLEALPQESWQNKHAGSNGNRATNGAPSLVSRGMQGASANTLNSETSTKRAAVYRMNRNHRGLCVIVNNHSFTSLKDRQGTHKDAEILSHVFQWLGFTVHIHNNVTKVEMEMVLQKQKCNPAHADGDCFVFCILTHGRFGAVYSSDEALIPIREIMSHFTALQCPRLAEKPKLFFIQACQGEEIQPSVSIEADALNPEQAPTSLQDSIPAEADFLLGLATVPGYVSFRHVEEGSWYIQSLCNHLKKLVPRMLKFLEKTMEIRGRKRTVWGAKQISATSLPTAISAQTPRPPMRRWSSVS 840348AE602B8243 cul3_human Cullin-3 CUL3 KIAA0617 protein protein human Homo sapiens Human MSNLSKGTGSRKDTKMRIRAFPMTMDEKYVNSIWDLLKNAIQEIQRKNNSGLSFEELYRNAYTMVLHKHGEKLYTGLREVVTEHLINKVREDVLNSLNNNFLQTLNQAWNDHQTAMVMIRDILMYMDRVYVQQNNVENVYNLGLIIFRDQVVRYGCIRDHLRQTLLDMIARERKGEVVDRGAIRNACQMLMILGLEGRSVYEEDFEAPFLEMSAEFFQMESQKFLAENSASVYIKKVEARINEEIERVMHCLDKSTEEPIVKVVERELISKHMKTIVEMENSGLVHMLKNGKTEDLGCMYKLFSRVPNGLKTMCECMSSYLREQGKALVSEEGEGKNPVDYIQGLLDLKSRFDRFLLESFNNDRLFKQTIAGDFEYFLNLNSRSPEYLSLFIDDKLKKGVKGLTEQEVETILDKAMVLFRFMQEKDVFERYYKQHLARRLLTNKSVSDDSEKNMISKLKTECGCQFTSKLEGMFRDMSISNTTMDEFRQHLQATGVSLGGVDLTVRVLTTGYWPTQSATPKCNIPPAPRHAFEIFRRFYLAKHSGRQLTLQHHMGSADLNATFYGPVKKEDGSEVGVGGAQVTGSNTRKHILQVSTFQMTILMLFNNREKYTFEEIQQETDIPERELVRALQSLACGKPTQRVLTKEPKSKEIENGHIFTVNDQFTSKLHRVKIQTVAAKQGESDPERKETRQKVDDDRKHEIEAAIVRIMKSRKKMQHNVLVAEVTQQLKARFLPSPVVIKKRIEGLIEREYLARTPEDRKVYTYVA DIP protein Q13618 original sequence version: 140 DIP protein Q13618 original sequence version: 163 DIP protein Q13618 original sequence version: 121 DIP protein Q13618 original sequence version: 161 A1A02022480BF099 ub2d1_human Ubiquitin-conjugating enzyme E2 D1 Ubiquitin-protein ligase D1 Ubiquitin carrier protein D1 UbcH5 Ubiquitin-conjugating enzyme E2-17 kDa 1 Ubiquitin-conjugating enzyme E2(17)KB 1 UBC4/5 homolog Stimulator of Fe transport UBE2D1 UBCH5 UBCH5A SFT UBC5A (E3-independent) E2 ubiquitin-conjugating enzyme D1 E2 ubiquitin-conjugating enzyme D1 protein protein human Homo sapiens Human MALKRIQKELSDLQRDPPAHCSAGPVGDDLFHWQATIMGPPDSAYQGGVFFLTVHFPTDYPFKPPKIAFTTKIYHPNINSNGSICLDILRSQWSPALTVSKVLLSICSLLCDPNPDDPLVPDIAQIYKSDKEKYNRHAREWTQKYAM DIP protein P51668 original sequence version: 143 DIP protein P51668 original sequence version: 119 DIP protein P51668 original sequence version: 107 2E96FD0179EE119D fadd_human FAS-associated death domain protein FAS-associating death domain-containing protein Mediator of receptor induced toxicity FADD MORT1 GIG3 Growth-inhibiting gene 3 protein protein protein human Homo sapiens Human MDPFLVLLHSVSSSLSSSELTELKFLCLGRVGKRKLERVQSGLDLFSMLLEQNDLEPGHTELLRELLASLRRHDLLRRVDDFEAGAAAGAAPGEEDLCAAFNVICDNVGKDWRRLARQLKVSDTKIDSIEDRYPRNLTERVRESLRIWKNTEKENATVAHLVGALRSCQMNLVADLVQEVQQARDLQNRSGAMSPMSWNSDASTSEAS DIP protein Q13158 original sequence version: 155 DIP protein Q13158 original sequence version: 130 DIP protein Q13158 original sequence version: 110 DIP protein Q13158 original sequence version: 118 DIP protein Q13158 original sequence version: 141 DIP protein Q13158 original sequence version: 166 0E65E2F852E83507 tnf10_human Tumor necrosis factor ligand superfamily member 10 TNF-related apoptosis-inducing ligand Apo-2 ligand TNFSF10 APO2L TRAIL protein protein human Homo sapiens Human MAMMEVQGGPSLGQTCVLIVIFTVLLQSLCVAVTYVYFTNELKQMQDKYSKSGIACFLKEDDSYWDPNDEESMNSPCWQVKWQLRQLVRKMILRTSEETISTVQEKQQNISPLVRERGPQRVAAHITGTRGRSNTLSSPNSKNEKALGRKINSWESSRSGHSFLSNLHLRNGELVIHEKGFYYIYSQTYFRFQEEIKENTKNDKQMVQYIYKYTSYPDPILLMKSARNSCWSKDAEYGLYSIYQGGIFELKENDRIFVSVTNEHLIDMDHEASFFGAFLVG DIP protein P50591 original sequence version: 150 DDAAAF78DAAB2F6D tr10a_human Tumor necrosis factor receptor superfamily member 10A Death receptor 4 TNF-related apoptosis-inducing ligand receptor 1 TNFRSF10A APO2 DR4 TRAILR1 protein protein human Homo sapiens Human MAPPPARVHLGAFLAVTPNPGSAASGTEAAAATPSKVWGSSAGRIEPRGGGRGALPTSMGQHGPSARARAGRAPGPRPAREASPRLRVHKTFKFVVVGVLLQVVPSSAATIKLHDQSIGTQQWEHSPLGELCPPGSHRSEHPGACNRCTEGVGYTNASNNLFACLPCTACKSDEEERSPCTTTRNTACQCKPGTFRNDNSAEMCRKCSRGCPRGMVKVKDCTPWSDIECVHKESGNGHNIWVILVVTLVVPLLLVAVLIVCCCIGSGCGGDPKCMDRVCFWRLGLLRGPGAEDNAHNEILSNADSLSTFVSEQQMESQEPADLTGVTVQSPGEAQCLLGPAEAEGSQRRRLLVPANGADPTETLMLFFDKFANIVPFDSWDQLMRQLDLTKNEIDVVRAGTAGPGDALYAMLMKWVNKTGRNASIHTLLDALERMEERHAREKIQDLLVDSGKFIYLEDGTGSAVSLE 7E96619D0BDC0CD4 otu7b_human OTU domain-containing protein 7B Zinc finger protein Cezanne Zinc finger A20 domain-containing protein 1 Cellular zinc finger anti-NF-kappa-B protein OTUD7B ZA20D1 protein protein human Homo sapiens Human MTLDMDAVLSDFVRSTGAEPGLARDLLEGKNWDVNAALSDFEQLRQVHAGNLPPSFSEGSGGSRTPEKGFSDREPTRPPRPILQRQDDIVQEKRLSRGISHASSSIVSLARSHVSSNGGGGGSNEHPLEMPICAFQLPDLTVYNEDFRSFIERDLIEQSMLVALEQAGRLNWWVSVDPTSQRLLPLATTGDGNCLLHAASLGMWGFHDRDLMLRKALYALMEKGVEKEALKRRWRWQQTQQNKESGLVYTEDEWQKEWNELIKLASSEPRMHLGTNGANCGGVESSEEPVYESLEEFHVFVLAHVLRRPIVVVADTMLRDSGGEAFAPIPFGGIYLPLEVPASQCHRSPLVLAYDQAHFSALVSMEQKENTKEQAVIPLTDSEYKLLPLHFAVDPGKGWEWGKDDSDNVRLASVILSLEVKLHLLHSYMNVKWIPLSSDAQAPLAQPESPTASAGDEPRSTPESGDSDKESVGSSSTSNEGGRRKEKSKRDREKDKKRADSVANKLGSFGKTLGSKLKKNMGGLMHSKGSKPGGVGTGLGGSSGTETLEKKKKNSLKSWKGGKEEAAGDGPVSEKPPAESVGNGGSKYSQEVMQSLSILRTAMQGEGKFIFVGTLKMGHRHQYQEEMIQRYLSDAEERFLAEQKQKEAERKIMNGGIGGGPPPAKKPEPDAREEQPTGPPAESRAMAFSTGYPGDFTIPRPSGGGVHCQEPRRQLAGGPCVGGLPPYATFPRQCPPGRPYPHQDSIPSLEPGSHSKDGLHRGALLPPPYRVADSYSNGYREPPEPDGWAGGLRGLPPTQTKCKQPNCSFYGHPETNNFCSCCYREELRRREREPDGELLVHRF DIP protein Q6GQQ9 original sequence version: 96 6D386C864B12EE57 cflar_human CASP8 and FADD-like apoptosis regulator Cellular FLICE-like inhibitory protein Caspase-eight-related protein Caspase-like apoptosis regulatory protein MACH-related inducer of toxicity Caspase homolog Inhibitor of FLICE FADD-like antiapoptotic molecule 1 Usurpin CFLAR CASH CLARP MRIT CASP8AP1 protein protein human Homo sapiens Human MSAEVIHQVEEALDTDEKEMLLFLCRDVAIDVVPPNVRDLLDILRERGKLSVGDLAELLYRVRRFDLLKRILKMDRKAVETHLLRNPHLVSDYRVLMAEIGEDLDKSDVSSLIFLMKDYMGRGKISKEKSFLDLVVELEKLNLVAPDQLDLLEKCLKNIHRIDLKTKIQKYKQSVQGAGTSYRNVLQAAIQKSLKDPSNNFRLHNGRSKEQRLKEQLGAQQEPVKKSIQESEAFLPQSIPEERYKMKSKPLGICLIIDCIGNETELLRDTFTSLGYEVQKFLHLSMHGISQILGQFACMPEHRDYDSFVCVLVSRGGSQSVYGVDQTHSGLPLHHIRRMFMGDSCPYLAGKPKMFFIQNYVVSEGQLEDSSLLEVDGPAMKNVEFKAQKRGLCTVHREADFFWSLCTADMSLLEQSHSSPSLYLQCLSQKLRQERKRPLLDLHIELNGYMYDWNSRVSAKEKYYVWLQHTLRKKLILSYT 8C6D7E92AE1EB672 ub2v1_human Ubiquitin-conjugating enzyme E2 variant 1 CROC-1 TRAF6-regulated IKK activator 1 beta Uev1A P/OKcl.19 UBE2V1 CROC1 UBE2V UEV1 protein protein human Homo sapiens Human MAATTGSGVKVPRNFRLLEELEEGQKGVGDGTVSWGLEDDEDMTLTRWTGMIIGPPRTIYENRIYSLKIECGPKYPEAPPFVRFVTKINMNGVNSSNGVVDPRAISVLAKWQNSYSIKVVLQELRRLMMSKENMKLPQPPEGQCYSN BA53837F21977B3F tr10b_human Tumor necrosis factor receptor superfamily member 10B Death receptor 5 TNF-related apoptosis-inducing ligand receptor 2 UNQ160/PRO186 TNFRSF10B DR5 KILLER TRAILR2 TRICK2 ZTNFR9 protein protein human Homo sapiens Human MEQRGQNAPAASGARKRHGPGPREARGARPGPRVPKTLVLVVAAVLLLVSAESALITQQDLAPQQRAAPQQKRSSPSEGLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRCDSGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSDIECVHKESGTKHSGEVPAVEETVTSSPGTPASPCSLSGIIIGVTVAAVVLIVAVFVCKSLLWKKVLPYLKGICSGGGGDPERVDRSSQRPGAEDNVLNEIVSILQPTQVPEQEMEVQEPAEPTGVNMLSPGESEHLLEPAEAERSQRRRLLVPANEGDPTETLRQCFDDFADLVPFDSWEPLMRKLGLMDNEIKVAKAEAAGHRDTLYTMLIKWVNKTGRDASVHTLLDALETLGERLAKQKIEDHLLSSGKFMYLEGNADSAMS 60358EAF2A835870 ubb_human Polyubiquitin-B UBB protein protein human Homo sapiens Human MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGC 33011162F1C48BB1 tnfrsf10a-tnfsf10-1 5 13 unspecified role unspecified role prey prey 20 unspecified role unspecified role prey prey 17 unspecified role unspecified role prey prey 22 unspecified role unspecified role prey prey 16 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 18 unspecified role unspecified role bait bait 14 unspecified role unspecified role prey prey 19 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey association association TNFSF10 (1 ug/ml) To identify which E3 ligase supports caspase-8 ubiquitination, we treated H460 cells with Apo2L/TRAIL for 2 hr at 4C and isolated the DISC from HMW SEC fractions by IP with DR4-specific or control antibody. Tandem mass spectrometry (MS/MS) revealed the expected DISC components, namely Apo2L/TRAIL, DR4, DR5, FADD, caspase-8, caspase-10, and FLIP, as well as another DISC-associated component-the E3 ubiquitin-ligase subunit CUL3 (Figure 2A). Fig. 2A, Supp. Fig. 8A tnfsf10-cul3-1 9 14 unspecified role unspecified role prey prey region (possible) neddylated lysine neddylated lysine undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 17 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 H460, Bax+/− HCT116, and HT29 cells were incubated with Flag-epitope-tagged Apo2L/TRAIL (1 μg/ml) at 4C. Cell lysates were subjected to DISC IP with anti-Flag M2 antibody, followed by IB with antibody to CUL3 (Nedd, Neddylated). Fig. 2C tnfsf10-cul3-2 7 14 unspecified role unspecified role prey prey region (possible) neddylated lysine neddylated lysine undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 17 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 H460, Bax+/− HCT116, and HT29 cells were incubated with Flag-epitope-tagged Apo2L/TRAIL (1 μg/ml) at 4C. Cell lysates were subjected to DISC IP with anti-Flag M2 antibody, followed by IB with antibody to CUL3 (Nedd, Neddylated). Fig. 2C casp8-cul3-1 8 14 unspecified role unspecified role prey prey region (possible) neddylated lysine neddylated lysine undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role bait bait physical association physical association IP of caspase-8 from H460 cells further confirmed the interaction with neddylated CUL3 (Figure 2D). Fig. 2D casp8-cul3-2 8 14 unspecified role unspecified role bait bait region (possible) neddylated lysine neddylated lysine undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role prey prey physical association physical association Moreover, IP of CUL3 demonstrated ligand-induced CUL3 association with caspase-8 in H460 and H2122 cells Fig. 2E tnfsf10-cul3-3 9 11 unspecified role unspecified role prey prey 22 unspecified role unspecified role prey prey 17 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 16 unspecified role unspecified role prey prey 18 unspecified role unspecified role prey prey association association CUL3 knockdown did not significantly alter recruitment to the DISC of DR4, DR5, FADD, or caspase-8 however, it markedly decreased the amount of ubiquitinated caspase-8 in the DISC. Fig. 3C casp8-cul3-3 1 14 unspecified role unspecified role ancillary ancillary human-hct116 Human colon carcinoma HCT116 cells hct116 Human colon carcinoma line HCT116 23 unspecified role unspecified role ancillary ancillary 11 enzyme target enzyme target substrate substrate neutral component neutral component n-terminal t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range in vitro In vitro 10 unspecified role unspecified role ancillary ancillary 21 unspecified role unspecified role ancillary ancillary 15 enzyme enzyme neutral component neutral component ubiquitination ubiquitination reaction ubiquitination Additional CUL3 binding proteins may have been present in the assay but were not identified. Fig. 4D Ubiquitination of CASP8 CASP8 is a non-cleavable mutant - details not given in paper. tnfrsf10a-sqstm1-1 8 18 unspecified role unspecified role bait bait 22 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey association association Fig. 6A casp8-sqstm1-1 8 12 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey association association Fig. 6A CUL3 knockdown attenuated the association of p62 with the DISC. casp8-tnfrsf10a-1 4 11 unspecified role unspecified role neutral component neutral component 18 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig. 7A-C, 7E Upon ligand stimulation at 4�C, a fraction of caspase-8 was recruited to the plasma membrane, where it strongly colocalized with DR4 TNFSF10 casp8-tnfrsf10a-2 4 12 unspecified role unspecified role neutral component neutral component 11 unspecified role unspecified role neutral component neutral component 18 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig. 7E, Supp. Fig. 12 Costaining for caspase-8, p62, and DR4 confirmed the presence of speckles containing (1) caspase-8 and p62 without DR4, (2) caspase-8 and DR4 without p62, and (3) caspase-8, DR4, and p62 (Figure 7E). TNFSF10 casp8-sqstm1-2 4 11 unspecified role unspecified role neutral component neutral component 12 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig. 7E, Supp. Fig. 12 Costaining for caspase-8, p62, and DR4 confirmed the presence of speckles containing (1) caspase-8 and p62 without DR4, (2) caspase-8 and DR4 without p62, and (3) caspase-8, DR4, and p62 (Figure 7E). TNFSF10 casp8-cul3-4 3 11 unspecified role unspecified role bait bait 14 unspecified role unspecified role prey prey region (possible) neddylated lysine neddylated lysine undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Apo2L/TRAIL induces association of CUL3 with C8. Supp. Fig. S4B TNFSF10 tnfrsf10b-tnfsf10-2 5 19 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey 16 unspecified role unspecified role prey prey 14 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 17 unspecified role unspecified role prey prey 13 unspecified role unspecified role prey prey 22 unspecified role unspecified role prey prey 20 unspecified role unspecified role prey prey 18 unspecified role unspecified role bait bait association association TNFSF10 (1 ug/ml) Supp. Fig. 8A casp8-otud7b-1 8 19 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association IP of caspase-8 from H460 cells further confirmed the interaction with neddylated CUL3 (Figure 2D). Fig. 2D TNFSF10 rps27a-sqstm1-3 4 12 unspecified role unspecified role neutral component neutral component 23 unspecified role unspecified role neutral component neutral component colocalization colocalization Supp. Fig. 11 After 48 hours, cells were fixed and stained with anti-p62 or anti-ubiquitin antibodies and analyzed by conforcal microscopy. TNFSF10 casp8-sqstm1-3 6 11 unspecified role unspecified role neutral component neutral component 12 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig. 6F resulting in colocalization of NCC8 and p62-positive aggregates casp8-cul3-5 2 14 unspecified role unspecified role prey prey region (possible) neddylated lysine neddylated lysine undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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 11 unspecified role unspecified role bait bait n-terminal t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Cotransfection of differentially tagged NCC8 and CUL3 versions followed by reciprocal IP and IB confirmed the ability of the two proteins to interact (Figures 2F and 2G). Fig. 2G CASP8 is a non-cleavable mutant - details not given in the paper. casp8-cul3-6 2 14 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 11 unspecified role unspecified role prey prey n-terminal t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Cotransfection of differentially tagged NCC8 and CUL3 versions followed by reciprocal IP and IB confirmed the ability of the two proteins to interact (Figures 2F and 2G). Fig. 2F CASP8 is a non-cleavable mutant - details not given in the paper.