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/ cho-2003-1 in vitro In vitro pull down pull down autoradiography autoradiography Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ. From PMID:11278289 (Fig.1A)ASK1 is inferred to be of human origin according to sequence length. From PMID:8137421 JNK1 is of mouse origin. ASK1 is inferred to be human according to protein length provided in Fig. 3.CIIA is inferred to be mouse according to protein length provided in Fig. 3. CAD is inferred to be mouse according to protein length provided in Fig. 4. Email sent to author (11/11/18) to request origin species of T7-ICAd and T7-ICAD-CAD, HA-ICAD-L, ICAD-L, Flag-GSTM1, Flag-Daxx, or TRAF2, no interaction detected with these constructs has been annotated so far. Kinase assays were not annotated because interactors were not sufficiently purified. ejchoi@korea.ac.kr The Journal of cell biology 2003 imex curation Only protein-protein interactions imex curation cho-2003-3 mouse-brain Mus musculus brain brain Brain [cerebrum] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ. From PMID:11278289 (Fig.1A)ASK1 is inferred to be of human origin according to sequence length. From PMID:8137421 JNK1 is of mouse origin. ASK1 is inferred to be human according to protein length provided in Fig. 3.CIIA is inferred to be mouse according to protein length provided in Fig. 3. CAD is inferred to be mouse according to protein length provided in Fig. 4. Email sent to author (11/11/18) to request origin species of T7-ICAd and T7-ICAD-CAD, HA-ICAD-L, ICAD-L, Flag-GSTM1, Flag-Daxx, or TRAF2, no interaction detected with these constructs has been annotated so far. Kinase assays were not annotated because interactors were not sufficiently purified. ejchoi@korea.ac.kr The Journal of cell biology 2003 imex curation Only protein-protein interactions imex curation cho-2003-4 mouse-embryonic_fibroblast Mus musculus embryonic fibroblasts embryonic_fibroblast Embryonic fibroblast Fibroblasts taken from an embryonic as opposed to a foetal or adult source. Foetus is defined as the stage at which tissue diferentiation is initiated. Tissue taken from an embryonic as opposed to a foetal or adult source. Embryo is defined as the development stage prior to organ formation. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ. From PMID:11278289 (Fig.1A)ASK1 is inferred to be of human origin according to sequence length. From PMID:8137421 JNK1 is of mouse origin. ASK1 is inferred to be human according to protein length provided in Fig. 3.CIIA is inferred to be mouse according to protein length provided in Fig. 3. CAD is inferred to be mouse according to protein length provided in Fig. 4. Email sent to author (11/11/18) to request origin species of T7-ICAd and T7-ICAD-CAD, HA-ICAD-L, ICAD-L, Flag-GSTM1, Flag-Daxx, or TRAF2, no interaction detected with these constructs has been annotated so far. Kinase assays were not annotated because interactors were not sufficiently purified. ejchoi@korea.ac.kr The Journal of cell biology 2003 imex curation Only protein-protein interactions imex curation cho-2003-5 mouse-l-929 Mus musculus L929 fibroblasts l-929 L-929 fibroblasts anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ. From PMID:11278289 (Fig.1A)ASK1 is inferred to be of human origin according to sequence length. From PMID:8137421 JNK1 is of mouse origin. ASK1 is inferred to be human according to protein length provided in Fig. 3.CIIA is inferred to be mouse according to protein length provided in Fig. 3. CAD is inferred to be mouse according to protein length provided in Fig. 4. Email sent to author (11/11/18) to request origin species of T7-ICAd and T7-ICAD-CAD, HA-ICAD-L, ICAD-L, Flag-GSTM1, Flag-Daxx, or TRAF2, no interaction detected with these constructs has been annotated so far. Kinase assays were not annotated because interactors were not sufficiently purified. ejchoi@korea.ac.kr The Journal of cell biology 2003 imex curation Only protein-protein interactions imex curation cho-2003-8 mouse-nih_3t3 Mus musculus Swiss NIH 3t3 embryo fibroblast cells nih_3t3 Mouse Swiss NIH embryo contact inhibited fibroblast cells confocal microscopy confocal microscopy anti-tag immunocytochemistry anti-tag immunocytochemistry Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ. From PMID:11278289 (Fig.1A)ASK1 is inferred to be of human origin according to sequence length. From PMID:8137421 JNK1 is of mouse origin. ASK1 is inferred to be human according to protein length provided in Fig. 3.CIIA is inferred to be mouse according to protein length provided in Fig. 3. CAD is inferred to be mouse according to protein length provided in Fig. 4. Email sent to author (11/11/18) to request origin species of T7-ICAd and T7-ICAD-CAD, HA-ICAD-L, ICAD-L, Flag-GSTM1, Flag-Daxx, or TRAF2, no interaction detected with these constructs has been annotated so far. Kinase assays were not annotated because interactors were not sufficiently purified. ejchoi@korea.ac.kr mouse adult brain cDNA The Journal of cell biology 2003 imex curation Only protein-protein interactions imex curation cho-2003-2 mouse-nih_3t3 Mus musculus Swiss NIH 3t3 embryo fibroblast cells nih_3t3 Mouse Swiss NIH embryo contact inhibited fibroblast cells density sedimentation cosedimentation through density gradient density sedimentation western blot western blot Immuno blot Immuno blot Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ. From PMID:11278289 (Fig.1A)ASK1 is inferred to be of human origin according to sequence length. From PMID:8137421 JNK1 is of mouse origin. ASK1 is inferred to be human according to protein length provided in Fig. 3.CIIA is inferred to be mouse according to protein length provided in Fig. 3. CAD is inferred to be mouse according to protein length provided in Fig. 4. Email sent to author (11/11/18) to request origin species of T7-ICAd and T7-ICAD-CAD, HA-ICAD-L, ICAD-L, Flag-GSTM1, Flag-Daxx, or TRAF2, no interaction detected with these constructs has been annotated so far. Kinase assays were not annotated because interactors were not sufficiently purified. ejchoi@korea.ac.kr The Journal of cell biology 2003 imex curation Only protein-protein interactions imex curation cho-2003-6 yeast Saccharomyces cerevisiae Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ. From PMID:11278289 (Fig.1A)ASK1 is inferred to be of human origin according to sequence length. From PMID:8137421 JNK1 is of mouse origin. ASK1 is inferred to be human according to protein length provided in Fig. 3.CIIA is inferred to be mouse according to protein length provided in Fig. 3. CAD is inferred to be mouse according to protein length provided in Fig. 4. Email sent to author (11/11/18) to request origin species of T7-ICAd and T7-ICAD-CAD, HA-ICAD-L, ICAD-L, Flag-GSTM1, Flag-Daxx, or TRAF2, no interaction detected with these constructs has been annotated so far. Kinase assays were not annotated because interactors were not sufficiently purified. ejchoi@korea.ac.kr mouse adult brain cDNA The Journal of cell biology 2003 imex curation Only protein-protein interactions imex curation cho-2003-7 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 Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ., Cho SG., Kim JW., Lee YH., Hwang HS., Kim MS., Ryoo K., Kim MJ., Noh KT., Kim EK., Cho JH., Yoon KW., Cho EG., Park HS., Chi SW., Lee MJ., Kang SS., Ichijo H., Choi EJ. From PMID:11278289 (Fig.1A)ASK1 is inferred to be of human origin according to sequence length. From PMID:8137421 JNK1 is of mouse origin. ASK1 is inferred to be human according to protein length provided in Fig. 3.CIIA is inferred to be mouse according to protein length provided in Fig. 3. CAD is inferred to be mouse according to protein length provided in Fig. 4. Email sent to author (11/11/18) to request origin species of T7-ICAd and T7-ICAD-CAD, HA-ICAD-L, ICAD-L, Flag-GSTM1, Flag-Daxx, or TRAF2, no interaction detected with these constructs has been annotated so far. Kinase assays were not annotated because interactors were not sufficiently purified. ejchoi@korea.ac.kr mouse adult brain cDNA The Journal of cell biology 2003 imex curation Only protein-protein interactions imex curation m3k5_human Mitogen-activated protein kinase kinase kinase 5 MAPK/ERK kinase kinase 5 Apoptosis signal-regulating kinase 1 MAP3K5 ASK1 MAPKKK5 MEKK5 protein protein human Homo sapiens Human MSTEADEGITFSVPPFAPSGFCTIPEGGICRRGGAAAVGEGEEHQLPPPPPGSFWNVESAAAPGIGCPAATSSSSATRGRGSSVGGGSRRTTVAYVINEASQGQLVVAESEALQSLREACETVGATLETLHFGKLDFGETTVLDRFYNADIAVVEMSDAFRQPSLFYHLGVRESFSMANNIILYCDTNSDSLQSLKEIICQKNTMCTGNYTFVPYMITPHNKVYCCDSSFMKGLTELMQPNFELLLGPICLPLVDRFIQLLKVAQASSSQYFRESILNDIRKARNLYTGKELAAELARIRQRVDNIEVLTADIVINLLLSYRDIQDYDSIVKLVETLEKLPTFDLASHHHVKFHYAFALNRRNLPGDRAKALDIMIPMVQSEGQVASDMYCLVGRIYKDMFLDSNFTDTESRDHGASWFKKAFESEPTLQSGINYAVLLLAAGHQFESSFELRKVGVKLSSLLGKKGNLEKLQSYWEVGFFLGASVLANDHMRVIQASEKLFKLKTPAWYLKSIVETILIYKHFVKLTTEQPVAKQELVDFWMDFLVEATKTDVTVVRFPVLILEPTKIYQPSYLSINNEVEEKTISIWHVLPDDKKGIHEWNFSASSVRGVSISKFEERCCFLYVLHNSDDFQIYFCTELHCKKFFEMVNTITEEKGRSTEEGDCESDLLEYDYEYDENGDRVVLGKGTYGIVYAGRDLSNQVRIAIKEIPERDSRYSQPLHEEIALHKHLKHKNIVQYLGSFSENGFIKIFMEQVPGGSLSALLRSKWGPLKDNEQTIGFYTKQILEGLKYLHDNQIVHRDIKGDNVLINTYSGVLKISDFGTSKRLAGINPCTETFTGTLQYMAPEIIDKGPRGYGKAADIWSLGCTIIEMATGKPPFYELGEPQAAMFKVGMFKVHPEIPESMSAEAKAFILKCFEPDPDKRACANDLLVDEFLKVSSKKKKTQPKLSALSAGSNEYLRSISLPVPVLVEDTSSSSEYGSVSPDTELKVDPFSFKTRAKSCGERDVKGIRTLFLGIPDENFEDHSAPPSPEEKDSGFFMLRKDSERRATLHRILTEDQDKIVRNLMESLAQGAEEPKLKWEHITTLIASLREFVRSTDRKIIATTLSKLKLELDFDSHGISQVQVVLFGFQDAVNKVLRNHNIKPHWMFALDSIIRKAVQTAITILVPELRPHFSLASESDTADQEDLDVEDDHEEQPSNQTVRRPQAVIEDAVATSGVSTLSSTVSHDSQSAHRSLNVQLGRMKIETNRLLEELVRKEKELQALLHRAIEEKDQEIKHLKLKSQPIEIPELPVFHLNSSGTNTEDSELTDWLRVNGADEDTISRFLAEDYTLLDVLYYVTRDDLKCLRLRGGMLCTLWKAIIDFRNKQT vps28_mouse Vacuolar protein sorting-associated protein 28 homolog Vps28 Caspase-activated DNase inhibitor that interacts with ASK1 ESCRT-I complex subunit VPS28 protein protein mouse Mus musculus Mouse MFHGIPATPGVGAPGNKPELYEEVKLYKNAREREKYDNMAELFAVVKTMQALEKAYIKDCVTPNEYTAACSRLLVQYKAAFRQVQGSEISSIDEFCRKFRLDCPLAMERIKEDRPITIKDDKGNLNRCIADVVSLFITVMDKLRLEIRAMDEIQPDLRELMETMHRMSHLPPDFEGRQTVSQWLQTLSGMSASDELDDSQVRQMLFDLESAYNAFNRFLHA dffb_mouse DNA fragmentation factor subunit beta Dffb Cad DNA fragmentation factor 40 kDa subunit Caspase-activated deoxyribonuclease protein protein mouse Mus musculus Mouse MCAVLRQPKCVKLRALHSACKFGVAARSCQELLRKGCVRFQLPMPGSRLCLYEDGTEVTDDCFPGLPNDAELLLLTAGETWHGYVSDITRFLSVFNEPHAGVIQAARQLLSDEQAPLRQKLLADLLHHVSQNITAETREQDPSWFEGLESRFRNKSGYLRYSCESRIRGYLREVSAYTSMVDEAAQEEYLRVLGSMCQKLKSVQYNGSYFDRGAEASSRLCTPEGWFSCQGPFDLESCLSKHSINPYGNRESRILFSTWNLDHIIEKKRTVVPTLAEAIQDGREVNWEYFYSLLFTAENLKLVHIACHKKTTHKLECDRSRIYRPQTGSRRKQPARKKRPARKR mint Nuclease that induces DNA fragmentation and chromatin condensation during apoptosis. Degrades naked DNA and induces apoptotic morphology homomint m3k5_mouse Mitogen-activated protein kinase kinase kinase 5 MAPK/ERK kinase kinase 5 Apoptosis signal-regulating kinase 1 Map3k5 Ask1 Mekk5 protein protein mouse Mus musculus Mouse MGTEAGEGITFSVPPFASVGFCTIPEGGSCRRGGGAATAAEGEPSLQPLLVPPPPPPPGSFWNVESAAAPGTSCPTTAPGSSATRGRGNSGSGGGRRTTVAYVINEASQGQLVVAESEALQSLREACEAVGATLETLHFGKLDFGETAVLDRFYNADIAVVEMSDAFRQPSLFYHLGVRESFSMANNIILYCDTNSDSLQSLKEIICQKNTVCTGNYTFIPYMVTPHNKVYCCDSSFMKGLTELMQPNFELLLGPICLPLVDRFVQLLKVAQASSSQYFRESILSDIRKARNLYTGKELAAELARIRQRVDNIEVLTADIVINLLLSYRDIQDYDSIVKLVETLEKLPTFDLASHHHVKFHYAFALNRRNLPGDRAKALDIMIPMVQSEEQVASDMYCLVGRIYKDMFLDSNFTDTESRDHGASWFKKAFESEPTLQSGINYAVLLLAAGHQFESSFELRKVGVKLSSLLGKKGNLEKLQSYWEVGFFLGASVLANDHLRVIQASEKLFRLKTPAWYLKSIVETILIYKHFVKLTTEQPSAKQELVDFWMDFLVEATKTDVTVVRFPVLILEPTKIYQPSYLSINNEVEEKTISIWHVLPDDKKGIHEWNFGASSVRGVSISKFEERCCFLYVLHNSDDFQIYFCTELHCKRFFEMVNTITEEKGRGAEDGDCEGDSLEYDYEYDENGDRVVLGKGTYGIVYAGRDLSNQVRIAIKEIPERDSRYSQPLHEEIALHKHLKHKNIVQYLGSFSENGFIKIFMEQVPGGSLSALLRSKWGPLKDNEQTIGFYTKQILEGLKYLHDNQIVHRDIKGDNVLINTYSGVLKISDFGTSKRLAGINPCTETFTGTLQYMAPEIIDKGPRGYGKAADIWSLGCTIIEMATGKPPFYELGEPQAAMFKVGMFKVHPEIPESMSAEAKAFILKCFEPDPDKRACANDLLIDEFLKVSSKKKKTQPKLSALSTGSNEYLRSISLPVPVLVEDTSSSSEYGSVSPDTELKADPFSFKARAKSCGEKDGKGIRTLFLGIPDENFEDHSAPPSPEEKDSGFFMLRKDSERRATLHRILTEDQDKVVRNLMESLAQGAEEPKLKWEHITTLISSLREFVRSTDRKIIATTLSKLKLELDFDSHGISQVQVVLFGFQDAVNKVLRNHNIKPHWMFALDSIIRKAVQTAITILVPELRPHFSLASESDTADPEDLDVEDEHEELSSNQTVRRPQAITEDAVATSGVSTLSSTVSHDSQNAHRSLNVQLGRMKIETNRLLEELVRKERELQALLHQAIEEKDQEIRHLKLKSQPIDIPGFPVCHLNSPGTTTEDSELPGWLRENGADEDTISRFLAEDYTLVDVLYYVTRDDLKCLRLRGGMLCTLWKAIIDFRNKC dffa_mouse DNA fragmentation factor subunit alpha Dffa Icad DNA fragmentation factor 45 kDa subunit Inhibitor of CAD protein protein mouse Mus musculus Mouse MELSRGASAPDPDDVRPLKPCLLRRNHSRDQHGVAASSLEELRSKACELLAIDKSLTPITLVLAEDGTIVDDDDYFLCLPSNTKFVALACNEKWIYNDSDGGTAWVSQESFEADEPDSRAGVKWKNVARQLKEDLSSIILLSEEDLQALIDIPCAELAQELCQSCATVQGLQSTLQQVLDQREEARQSKQLLELYLQALEKEGNILSNQKESKAALSEELDAVDTGVGREMASEVLLRSQILTTLKEKPAPELSLSSQDLESVSKEDPKALAVALSWDIRKAETVQQACTTELALRLQQVQSLHSLRNLSARRSPLPGEPQRPKRAKRDSS ciia-ask1-2 3494210 N/A 3494218 unspecified role unspecified role prey prey cell lysate cell lysate in vitro translated in vitro translated protein in vitro translated 35s radiolabel 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes N/A 3494219 unspecified role unspecified role bait bait purified purified gst tag 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig. 3A vps28-map3k5-1 3494210 N/A 3494218 unspecified role unspecified role prey prey cell lysate cell lysate in vitro translated in vitro translated protein in vitro translated region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 35s radiolabel 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes N/A 3494219 unspecified role unspecified role bait bait purified purified gst tag 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 physical association physical association false false false Fig. 3B vps28-map3k5-4 3494210 N/A 3494220 unspecified role unspecified role prey prey cell lysate cell lysate in vitro translated in vitro translated protein in vitro translated 35s radiolabel 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes N/A 3494219 unspecified role unspecified role bait bait purified purified gst tag 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig. 4A vps28-map3k5-5 3494210 N/A 3494220 unspecified role unspecified role prey prey cell lysate cell lysate in vitro translated in vitro translated protein in vitro translated region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 35s radiolabel 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes N/A 3494219 unspecified role unspecified role bait bait purified purified gst tag 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 physical association physical association false false false Fig. 4B ask1-ciia-1 3494211 N/A 3494221 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3494219 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Fig.3C vps28-map3k5-7 3494211 N/A 3494220 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3494219 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Fig.4E vps28-map3k5-2 3494212 N/A 3494221 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3494219 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Fig.3C vps28-map3k5-6 3494212 N/A 3494220 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3494219 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Fig.4E vps28-map3k5-3 3494213 N/A 3494221 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3494219 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Fig.3E Subsequently, we examined a time course of the H2O2 action on the physical association of endogenous CIIA and ASK1 proteins in L929 cells (Fig. 3 E). Coimmunoprecipitation results indicated that H2O2-induced enhancement of the interaction between the two endogenous proteins was maximal at 1 h. vps28-map3k5-8 3494213 N/A 3494220 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3494219 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Fig.3E Physical interaction between CIIA and CAD was maximal after 30 min of an exposure of L929 cells to H2O2 (Fig. 4 F). vps28-map3k5-9 3494214 N/A 3494218 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell flag tag 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 3494219 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Fig. S1B map3k5-vps28-2 3494214 N/A 3494220 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell flag tag 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 3494219 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Fig. S1B Double-labeled immunostaining data revealed that ectopic CIIA was located in both the nucleus and the cytoplasm and that its distribution was overlapped with that of CAD or ASK1 ciia-ask1-1 3494215 N/A 3494220 unspecified role unspecified role neutral component neutral component living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3494222 unspecified role unspecified role neutral component neutral component living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3494221 unspecified role unspecified role neutral component neutral component living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3494219 anti tag western anti tag western blot anti tag western unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed living cell living cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Fig. 2 Immunoblot analysis of the glycerol gradient fractions revealed that the majority of ASK1 and CIIA were present in the same fractions (Fig. 2, fractions 10–16). CIIA and CAD also resided in the same fractions (Fig. 2, fractions 10–17). These fractionation results suggest that CIIA is in close proximity to ASK1 and to CAD. ask1-ciia-2 3494216 N/A 3494219 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed N/A 3494218 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association false false false Fig. 1A map3k5-vps28-1 3494217 N/A 3494219 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3494218 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed flag tag 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 physical association physical association false false false Fig. 3D Physical association between CIIA and ASK1 was also confirmed by coimmunoprecipitation in 293T cells transfected with plasmids encoding HA-tagged CIIA and Flag-tagged ASK1 (ASK1-Flag; Fig. 3 D). Interestingly, the interaction between ectopic CIIA and ASK1 was increased by H2O2 treatment.