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/ Physical and functional interactions between Daxx and STAT3. Physical and functional interactions between Daxx and STAT3. Oncogene (0950-9232) 2006 imex curation Muromoto R., Nakao K., Watanabe T., Sato N., Sekine Y., Sugiyama K., Oritani K., Shimoda K., Matsuda T. BioCreative - Critical Assessment of Information Extraction systems in Biology Apoptosis - Interactions involving proteins with a function related to apoptosis tmatsuda@pharm.hokudai.ac.jp 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions human-hep_3b Homo sapiens hepatocellular carcinoma (HCC) cells (p53-null). hep_3b human hepatocellular carcinoma (HCC) cells (p53-null). confocal microscopy confocal microscopy immunostaining immunostaining Muromoto R., Nakao K., Watanabe T., Sato N., Sekine Y., Sugiyama K., Oritani K., Shimoda K., Matsuda T. BioCreative - Critical Assessment of Information Extraction systems in Biology tmatsuda@pharm.hokudai.ac.jp Accepted 2006-SEP-13 by JYOTI. 24-JAN-2007: Contacted by JYOTI. Oncogene (0950-9232) 2006 Only protein-protein interactions imex curation Physical and functional interactions between Daxx and STAT3. Physical and functional interactions between Daxx and STAT3. Oncogene (0950-9232) 2006 imex curation Muromoto R., Nakao K., Watanabe T., Sato N., Sekine Y., Sugiyama K., Oritani K., Shimoda K., Matsuda T. BioCreative - Critical Assessment of Information Extraction systems in Biology Apoptosis - Interactions involving proteins with a function related to apoptosis tmatsuda@pharm.hokudai.ac.jp 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions 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. pull down pull down anti tag western anti tag western blot anti tag western Muromoto R., Nakao K., Watanabe T., Sato N., Sekine Y., Sugiyama K., Oritani K., Shimoda K., Matsuda T. BioCreative - Critical Assessment of Information Extraction systems in Biology tmatsuda@pharm.hokudai.ac.jp Accepted 2006-SEP-13 by JYOTI. 24-JAN-2007: Contacted by JYOTI. Oncogene (0950-9232) 2006 Only protein-protein interactions imex curation Physical and functional interactions between Daxx and STAT3. Physical and functional interactions between Daxx and STAT3. Oncogene (0950-9232) 2006 imex curation Muromoto R., Nakao K., Watanabe T., Sato N., Sekine Y., Sugiyama K., Oritani K., Shimoda K., Matsuda T. BioCreative - Critical Assessment of Information Extraction systems in Biology Apoptosis - Interactions involving proteins with a function related to apoptosis tmatsuda@pharm.hokudai.ac.jp 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions 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 Muromoto R., Nakao K., Watanabe T., Sato N., Sekine Y., Sugiyama K., Oritani K., Shimoda K., Matsuda T. BioCreative - Critical Assessment of Information Extraction systems in Biology tmatsuda@pharm.hokudai.ac.jp Accepted 2006-SEP-13 by JYOTI. Oncogene (0950-9232) 2006 Only protein-protein interactions imex curation daxx_human Death domain-associated protein 6 DAXX BING2 DAP6 Daxx Fas death domain-associated protein ETS1-associated protein 1 protein protein human Homo sapiens Human MATANSIIVLDDDDEDEAAAQPGPSHPLPNAASPGAEAPSSSEPHGARGSSSSGGKKCYKLENEKLFEEFLELCKMQTADHPEVVPFLYNRQQRAHSLFLASAEFCNILSRVLSRARSRPAKLYVYINELCTVLKAHSAKKKLNLAPAATTSNEPSGNNPPTHLSLDPTNAENTASQSPRTRGSRRQIQRLEQLLALYVAEIRRLQEKELDLSELDDPDSAYLQEARLKRKLIRLFGRLCELKDCSSLTGRVIEQRIPYRGTRYPEVNRRIERLINKPGPDTFPDYGDVLRAVEKAAARHSLGLPRQQLQLMAQDAFRDVGIRLQERRHLDLIYNFGCHLTDDYRPGVDPALSDPVLARRLRENRSLAMSRLDEVISKYAMLQDKSEEGERKKRRARLQGTSSHSADTPEASLDSGEGPSGMASQGCPSASRAETDDEDDEESDEEEEEEEEEEEEEATDSEEEEDLEQMQEGQEDDEEEDEEEEAAAGKDGDKSPMSSLQISNEKNLEPGKQISRSSGEQQNKGRIVSPSLLSEEPLAPSSIDAESNGEQPEELTLEEESPVSQLFELEIEALPLDTPSSVETDISSSRKQSEEPFTTVLENGAGMVSSTSFNGGVSPHNWGDSGPPCKKSRKEKKQTGSGPLGNSYVERQRSVHEKNGKKICTLPSPPSPLASLAPVADSSTRVDSPSHGLVTSSLCIPSPARLSQTPHSQPPRPGTCKTSVATQCDPEEIIVLSDSD 1B309ADDAA878040 stat3_human Signal transducer and activator of transcription 3 Acute-phase response factor STAT3 APRF protein protein human Homo sapiens Human MAQWNQLQQLDTRYLEQLHQLYSDSFPMELRQFLAPWIESQDWAYAASKESHATLVFHNLLGEIDQQYSRFLQESNVLYQHNLRRIKQFLQSRYLEKPMEIARIVARCLWEESRLLQTAATAAQQGGQANHPTAAVVTEKQQMLEQHLQDVRKRVQDLEQKMKVVENLQDDFDFNYKTLKSQGDMQDLNGNNQSVTRQKMQQLEQMLTALDQMRRSIVSELAGLLSAMEYVQKTLTDEELADWKRRQQIACIGGPPNICLDRLENWITSLAESQLQTRQQIKKLEELQQKVSYKGDPIVQHRPMLEERIVELFRNLMKSAFVVERQPCMPMHPDRPLVIKTGVQFTTKVRLLVKFPELNYQLKIKVCIDKDSGDVAALRGSRKFNILGTNTKVMNMEESNNGSLSAEFKHLTLREQRCGNGGRANCDASLIVTEELHLITFETEVYHQGLKIDLETHSLPVVVISNICQMPNAWASILWYNMLTNNPKNVNFFTKPPIGTWDQVAEVLSWQFSSTTKRGLSIEQLTTLAEKLLGPGVNYSGCQITWAKFCKENMAGKGFSFWVWLDNIIDLVKKYILALWNEGYIMGFISKERERAILSTKPPGTFLLRFSESSKEGGVTFTWVEKDISGKTQIQSVEPYTKQQLNNMSFAEIIMGYKIMDATNILVSPLVYLYPDIPKEEAFGKYCRPESQEHPEADPGSAAPYLKTKFICVTPTTCSNTIDLPMSPRTLDSLMQFGNNGEGAEPSAGGQFESLTFDMELTSECATSPM 6C00632211C8012D DIP protein P40763 original sequence version: 157 DIP protein P40763-1 original sequence version: 147 DIP protein P40763-1 original sequence version: 135 DIP protein P40763-1 original sequence version: 157 DIP protein P40763 original sequence version: 165 DIP protein P40763 original sequence version: 182 DIP protein P40763 original sequence version: 135 6C00632211C8012D daxx-stat3 3 4 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 5 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association As shown in Figure 3a, STAT3 interacted with Daxx in transient transfected 293T cells. 3 A The authors do not specify the species of the sequence of STAT3 used. The protein they use is 750 residues but the human protein (and mouse) is 770 residues. It is inferred that the potein is human as they are working with human cell lines. daxx-stat3-1 2 4 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 5 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 region STAT3 DNA-binding domain (320-493) binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain physical association physical association To delineate the domains in STAT3 that mediate the protein�protein interactions between STAT3 and Daxx, pull-down experiments were performed with a series of mutant GST-STAT3 proteins. As shown in Figure 3b, only the DNA-binding domain of STAT3 (320�493) showed a significant interaction with Daxx. 3 B The authors do not specify the species of the sequence of STAT3 used. The protein they use is 750 residues but the human protein (and mouse) is 770 residues. It is inferred that the potein is human as they are working with human cell lines. daxx-stat3-2 2 4 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 n-terminal N-terminal domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region central domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 5 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 region STAT3 DNA-binding domain (320-493) sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association As shown in Figure 3c, the N-terminal domain of Daxx interacted with the DNA-binding domain of STAT3, while the central domain of Daxx showed weak binding to that of STAT3 3 C The authors do not specify the species of the sequence of STAT3 used. The protein they use is 750 residues but the human protein (and mouse) is 770 residues. It is inferred that the potein is human as they are working with human cell lines. The N-terminal domain of Daxx interacted with the DNA-binding domain of STAT3, while the central domain of Daxx showed weak binding to that of STAT3. These data suggest that the DNA binding domain of STAT3 is required for the interaction with the N-terminal domain of Daxx. daxx-stat3-3 1 5 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role 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 colocalization colocalization STAT3 translocated into the nucleus and co-localized with Daxx in the nucleus after IL-6 stimulation (Figure 3d). Moreover, we could observe co-localization of endogenous Daxx and STAT3 in the nucleus 3 D, E The authors do not specify the species of the sequence of STAT3 used. The protein they use is 750 residues but the human protein (and mouse) is 770 residues. It is inferred that the potein is human as they are working with human cell lines. IL-6 treatment of the cells stimulates this colocalisation. These results suggest that activated STAT3 interacts with Daxx in the nucleus.