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/ Adenovirus E1B 55-kilodalton oncoprotein binds to Daxx and eliminates enhancement of p53-dependent transcription by Daxx. Adenovirus E1B 55-kilodalton oncoprotein binds to Daxx and eliminates enhancement of p53-dependent transcription by Daxx. J. Virol. (0022-538X) 2003 Zhao LY., Colosimo AL., Liu Y., Wan Y., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Source of protein Daxx taken from Invitrogen website (only provide human Daxx cDNA clone). dliao@ufl.edu 09-April-2009: Contacted by JYOTI. A HeLa cDNA library was used. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zhao LY., Colosimo AL., Liu Y., Wan Y., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Source of protein Daxx taken from Invitrogen website (only provide human Daxx cDNA clone). dliao@ufl.edu Accepted 2008-DEC-17 by ORCHARD. 09-April-2009: Contacted by JYOTI. A HeLa cDNA library was used. J. Virol. (0022-538X) 2003 Adenovirus E1B 55-kilodalton oncoprotein binds to Daxx and eliminates enhancement of p53-dependent transcription by Daxx. Adenovirus E1B 55-kilodalton oncoprotein binds to Daxx and eliminates enhancement of p53-dependent transcription by Daxx. J. Virol. (0022-538X) 2003 Zhao LY., Colosimo AL., Liu Y., Wan Y., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Source of protein Daxx taken from Invitrogen website (only provide human Daxx cDNA clone). dliao@ufl.edu 09-April-2009: Contacted by JYOTI. A HeLa cDNA library was used. in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zhao LY., Colosimo AL., Liu Y., Wan Y., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Source of protein Daxx taken from Invitrogen website (only provide human Daxx cDNA clone). dliao@ufl.edu Accepted 2008-DEC-17 by ORCHARD. 09-April-2009: Contacted by JYOTI. A HeLa cDNA library was used. J. Virol. (0022-538X) 2003 Adenovirus E1B 55-kilodalton oncoprotein binds to Daxx and eliminates enhancement of p53-dependent transcription by Daxx. Adenovirus E1B 55-kilodalton oncoprotein binds to Daxx and eliminates enhancement of p53-dependent transcription by Daxx. J. Virol. (0022-538X) 2003 Zhao LY., Colosimo AL., Liu Y., Wan Y., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Source of protein Daxx taken from Invitrogen website (only provide human Daxx cDNA clone). dliao@ufl.edu 09-April-2009: Contacted by JYOTI. A HeLa cDNA library was used. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Zhao LY., Colosimo AL., Liu Y., Wan Y., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Source of protein Daxx taken from Invitrogen website (only provide human Daxx cDNA clone). dliao@ufl.edu Accepted 2008-DEC-17 by ORCHARD. 09-April-2009: Contacted by JYOTI. A HeLa cDNA library was used. J. Virol. (0022-538X) 2003 Adenovirus E1B 55-kilodalton oncoprotein binds to Daxx and eliminates enhancement of p53-dependent transcription by Daxx. Adenovirus E1B 55-kilodalton oncoprotein binds to Daxx and eliminates enhancement of p53-dependent transcription by Daxx. J. Virol. (0022-538X) 2003 Zhao LY., Colosimo AL., Liu Y., Wan Y., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Source of protein Daxx taken from Invitrogen website (only provide human Daxx cDNA clone). dliao@ufl.edu 09-April-2009: Contacted by JYOTI. A HeLa cDNA library was used. yeasx Saccharomyces cerevisiae (Baker's yeast) 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 Zhao LY., Colosimo AL., Liu Y., Wan Y., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Source of protein Daxx taken from Invitrogen website (only provide human Daxx cDNA clone). dliao@ufl.edu Accepted 2008-DEC-17 by ORCHARD. 09-April-2009: Contacted by JYOTI. A HeLa cDNA library was used. J. Virol. (0022-538X) 2003 Adenovirus E1B 55-kilodalton oncoprotein binds to Daxx and eliminates enhancement of p53-dependent transcription by Daxx. Adenovirus E1B 55-kilodalton oncoprotein binds to Daxx and eliminates enhancement of p53-dependent transcription by Daxx. J. Virol. (0022-538X) 2003 Zhao LY., Colosimo AL., Liu Y., Wan Y., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Source of protein Daxx taken from Invitrogen website (only provide human Daxx cDNA clone). dliao@ufl.edu 09-April-2009: Contacted by JYOTI. A HeLa cDNA library was used. yeasx Saccharomyces cerevisiae (Baker's yeast) 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 predetermined predetermined participant predetermined Zhao LY., Colosimo AL., Liu Y., Wan Y., Liao D. Apoptosis - Interactions involving proteins with a function related to apoptosis Source of protein Daxx taken from Invitrogen website (only provide human Daxx cDNA clone). dliao@ufl.edu Accepted 2008-DEC-17 by ORCHARD. 09-April-2009: Contacted by JYOTI. A HeLa cDNA library was used. J. Virol. (0022-538X) 2003 p53_human Cellular tumor antigen p53 TP53 P53 Tumor suppressor p53 Phosphoprotein p53 Antigen NY-CO-13 protein protein human Homo sapiens Human MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLSPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYKQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPPEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMFRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD DIP protein P04637 original sequence version: 121 DIP protein P04637 original sequence version: 115 DIP protein P04637-1 original sequence version: 224 DIP protein P04637 original sequence version: 117 DIP protein P04637-1 original sequence version: 226 DIP protein P04637 original sequence version: 226 DIP protein P04637 original sequence version: 212 DIP protein P04637 original sequence version: 220 DIP protein P04637-1 original sequence version: 220 DIP protein P04637-1 original sequence version: 145 DIP protein P04637 original sequence version: 146 DIP protein P04637 original sequence version: 145 DIP protein P04637 original sequence version: 166 DIP protein P04637-1 original sequence version: 166 DIP protein P04637 original sequence version: 169 DIP protein P04637 original sequence version: 177 DIP protein P04637 original sequence version: 175 DIP protein P04637-1 original sequence version: 175 DIP protein P04637-1 original sequence version: 172 DIP protein P04637-1 original sequence version: 169 DIP protein P04637 original sequence version: 170 DIP protein P04637 original sequence version: 172 DIP protein P04637-1 original sequence version: 194 DIP protein P04637 original sequence version: 196 DIP protein P04637 original sequence version: 185 DIP protein P04637-1 original sequence version: 191 DIP protein P04637 original sequence version: 191 DIP protein P04637 original sequence version: 180 DIP protein P04637-1 original sequence version: 197 DIP protein P04637 original sequence version: 183 DIP protein P04637-1 original sequence version: 183 DIP protein P04637-1 original sequence version: 185 DIP protein P04637 original sequence version: 199 DIP protein P04637 original sequence version: 201 DIP protein P04637 original sequence version: 189 DIP protein P04637 original sequence version: 235 DIP protein P04637 original sequence version: 230 DIP protein P04637 original sequence version: 234 DIP protein P04637-1 original sequence version: 234 DIP protein P04637-1 original sequence version: 235 DIP protein P04637 original sequence version: 239 DIP protein P04637-1 original sequence version: 212 DIP protein P04637-1 original sequence version: 213 DIP protein P04637 original sequence version: 213 DIP protein P04637 original sequence version: 206 DIP protein P04637-1 original sequence version: 188 DIP protein P04637-1 original sequence version: 228 DIP protein P04637 original sequence version: 208 DIP protein P04637 original sequence version: 240 DIP protein P04637-1 original sequence version: 240 DIP protein P04637 original sequence version: 204 DIP protein P04637-1 original sequence version: 204 DIP protein P04637-1 original sequence version: 209 DIP protein P04637-1 original sequence version: 203 DIP protein P04637 original sequence version: 221 DIP protein P04637-1 original sequence version: 152 DIP protein P04637 original sequence version: 217 DIP protein P04637-1 original sequence version: 140 DIP protein P04637 original sequence version: 215 DIP protein P04637 original sequence version: 214 DIP protein P04637-1 original sequence version: 182 DIP protein P04637 original sequence version: 182 DIP protein P04637 original sequence version: 188 DIP protein P04637 original sequence version: 192 DIP protein P04637-1 original sequence version: 192 DIP protein P04637 original sequence version: 218 DIP protein P04637-1 original sequence version: 218 AD5C149FD8106131 e1b55_ade02 E1B 55 kDa protein E1B E1B-495R E1B protein, large T-antigen protein protein ade02 Human adenovirus C serotype 2 (HAdV-2) Human adenovirus 2 HAdV-2 MERRNPSERGVPAGFSGHASVESGGETQESPATVVFRPPGNNTDGGATAGGSQAAAAAGAEPMEPESRPGPSGMNVVQVAELFPELRRILTINEDGQGLKGVKRERGASEATEEARNLTFSLMTRHRPECVTFQQIKDNCANELDLLAQKYSIEQLTTYWLQPGDDFEEAIRVYAKVALRPDCKYKISKLVNIRNCCYISGNGAEVEIDTEDRVAFRCSMINMWPGVLGMDGVVIMNVRFTGPNFSGTVFLANTNLILHGVSFYGFNNTCVEAWTDVRVRGCAFYCCWKGVVCRPKSRASIKKCLFERCTLGILSEGNSRVRHNVASDCGCFMLVKSVAVIKHNMVCGNCEDRASQMLTCSDGNCHLLKTIHVASHSRKAWPVFEHNILTRCSLHLGNRRGVFLPYQCNLSHTKILLEPESMSKVNLNGVFDMTMKIWKVLRYDETRTRCRPCECGGKHIRNQPVMLDVTEELRPDHLVLACTRAEFGSSDEDTD D6665B27183720CC e1b55_ade12 E1B 55 kDa protein E1B protein, large T-antigen E1B-495R protein protein ade12 Human adenovirus A serotype 12 (HAdV-12) Human adenovirus 12 MEREIPPELGLHAGLHVNAAVEGMAEEEGLHLLAGAAFDHAAAADVARGEGGGAEPCGGGEVNMEQQVQEGHVLDSGEGPSCADDRDKQEKKESLKEAAVLSRLTVNLMSRPRLETVYWQELQDEFQRGDMHLQYKYSFEQLKTHWLEPWEDMECAIKAFAKLALRPDCSYRITKTVTITSCAYIIGNGAIVEVDTSDRVAFRCRMQGMGPGVVGLDGITFINVRFAGDKFKGIMFEANTCLVLHGVYFLNFSNICVESWNKVSARGCTFYGCWKGLVGRPKSKLSVKKCLFEKCVLALIVEGDAHIRHNAASENACFVLLKGMAILKHNMVCGVSDQTMRRFVTCADGNCHTLKTVHIVSHSRHCWPVCDHNMFMRCTIHLGLRRGMFRPSQCNFSHSNIMLEPEVFSRVCLNGVFDLSVELCKVIRYNDDTRHRCRQCECGSSHLELRPIVLNVTEELRSDHLTLSCLRTDYESSDEDDN 359E8082B7EF3659 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 e1b55_ade05 E1B 55 kDa protein E1B protein, large T-antigen E1B-495R protein protein ade05 Human adenovirus C serotype 5 (HAdV-5) HAdV-5 Human adenovirus 5 MERRNPSERGVPAGFSGHASVESGCETQESPATVVFRPPGDNTDGGAAAAAGGSQAAAAGAEPMEPESRPGPSGMNVVQVAELYPELRRILTITEDGQGLKGVKRERGACEATEEARNLAFSLMTRHRPECITFQQIKDNCANELDLLAQKYSIEQLTTYWLQPGDDFEEAIRVYAKVALRPDCKYKISKLVNIRNCCYISGNGAEVEIDTEDRVAFRCSMINMWPGVLGMDGVVIMNVRFTGPNFSGTVFLANTNLILHGVSFYGFNNTCVEAWTDVRVRGCAFYCCWKGVVCRPKSRASIKKCLFERCTLGILSEGNSRVRHNVASDCGCFMLVKSVAVIKHNMVCGNCEDRASQMLTCSDGNCHLLKTIHVASHSRKAWPVFEHNILTRCSLHLGNRRGVFLPYQCNLSHTKILLEPESMSKVNLNGVFDMTMKIWKVLRYDETRTRCRPCECGGKHIRNQPVMLDVTEELRPDHLVLACTRAEFGSSDEDTD 31EE3E4B9762DCE8 e1bl-daxx 4 7 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 9 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 15 13 physical association physical association 1 A With Ad2 E1B aa 155 to 495 as bait, we have screened approximately 106 yeast transformants. Among 11 positive clones, two identical clones encoding Daxx C-terminal aa 621 to 740 were isolated. e1bl-daxx-1 5 9 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 20 18 physical association physical association 1 B this Daxx fragment bound to Ad2 E1B aa 155 to 495 (sector 2) and the full-length Ad12 E1B protein (sector 6), as well as the Ad2 C-terminal aa 437 to 495 fragment e1bl-daxx-2 5 8 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association 1 B, D, E Full-length Daxx (Fig. 1C and D, sector 2), aa 1 to 574 (sector 3), and aa 516 to 574 (sector 6), as well as aa 621 to 740 (see Fig. 1A) bind to both the Ad2 and Ad12 E1B 55-kDa proteins. e1bl-daxx-3 5 7 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association 1 C, E Full-length Daxx (Fig. 1C and D, sector 2), aa 1 to 574 (sector 3), and aa 516 to 574 (sector 6), as well as aa 621 to 740 (see Fig. 1A) bind to both the Ad2 and Ad12 E1B 55-kDa proteins. e1bl-daxx-4 1 10 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association 2 A Daxx was present in the anti-E1B immunocomplexes daxx-e1bl 1 10 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association 2 B In a reciprocal IP, the E1B 55-kDa protein was precipitated by anti-Daxx M112 antibody e1bl-daxx-5 2 7 unspecified role unspecified role bait bait spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 9 unspecified role unspecified role prey prey spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection direct interaction direct interaction 2 C As shown in Fig. 2C, Daxx was coprecipitated with the E1B 55-kDa protein, indicating a direct interaction in vitro. e1bl-daxx-6 5 9 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association 1 B e1bl-daxx-7 3 10 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization 2 A Daxx was present in the anti-E1B immunocomplexes e1bl-daxx-8 3 6 unspecified role unspecified role neutral component neutral component 10 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization 3C In addition, simultaneous staining of the transfected 293 cells with goat anti-p53 antibody and 2A6 indicated that Daxx, p53, and the E1B 55-kDa protein were present in the same complex in the nuclear dots (Fig. 3C).