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/ Aprataxin, a novel protein that protects against genotoxic stress. Aprataxin, a novel protein that protects against genotoxic stress. Hum. Mol. Genet. (0964-6906) 2004 Gueven N., Becherel OJ., Kijas AW., Chen P., Howe O., Rudolph JH., Gatti R., Date H., Onodera O., Taucher-Scholz G., Lavin MF. Apoptosis - Interactions involving proteins with a function related to apoptosis Chromatin - Epigenetic interactions resulting in chromatin modulation nurig@qimr.edu.au 27-Aug-2008: Contacted by JYOTI. davet human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Gueven N., Becherel OJ., Kijas AW., Chen P., Howe O., Rudolph JH., Gatti R., Date H., Onodera O., Taucher-Scholz G., Lavin MF. Apoptosis - Interactions involving proteins with a function related to apoptosis nurig@qimr.edu.au Accepted 2008-SEP-13 by DAVET. 27-Aug-2008: Contacted by JYOTI. Hum. Mol. Genet. (0964-6906) 2004 davet Aprataxin, a novel protein that protects against genotoxic stress. Aprataxin, a novel protein that protects against genotoxic stress. Hum. Mol. Genet. (0964-6906) 2004 Gueven N., Becherel OJ., Kijas AW., Chen P., Howe O., Rudolph JH., Gatti R., Date H., Onodera O., Taucher-Scholz G., Lavin MF. Apoptosis - Interactions involving proteins with a function related to apoptosis Chromatin - Epigenetic interactions resulting in chromatin modulation nurig@qimr.edu.au 27-Aug-2008: Contacted by JYOTI. davet human-at3abr Homo sapiens lymphoblastoid cell line at3abr at3abr lymphoblastoid cell line anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Gueven N., Becherel OJ., Kijas AW., Chen P., Howe O., Rudolph JH., Gatti R., Date H., Onodera O., Taucher-Scholz G., Lavin MF. Apoptosis - Interactions involving proteins with a function related to apoptosis nurig@qimr.edu.au Accepted 2008-SEP-13 by DAVET. 27-Aug-2008: Contacted by JYOTI. Hum. Mol. Genet. (0964-6906) 2004 davet Aprataxin, a novel protein that protects against genotoxic stress. Aprataxin, a novel protein that protects against genotoxic stress. Hum. Mol. Genet. (0964-6906) 2004 Gueven N., Becherel OJ., Kijas AW., Chen P., Howe O., Rudolph JH., Gatti R., Date H., Onodera O., Taucher-Scholz G., Lavin MF. Apoptosis - Interactions involving proteins with a function related to apoptosis Chromatin - Epigenetic interactions resulting in chromatin modulation nurig@qimr.edu.au 27-Aug-2008: Contacted by JYOTI. davet in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Gueven N., Becherel OJ., Kijas AW., Chen P., Howe O., Rudolph JH., Gatti R., Date H., Onodera O., Taucher-Scholz G., Lavin MF. Apoptosis - Interactions involving proteins with a function related to apoptosis nurig@qimr.edu.au Accepted 2008-SEP-13 by DAVET. Hum. Mol. Genet. (0964-6906) 2004 davet Aprataxin, a novel protein that protects against genotoxic stress. Aprataxin, a novel protein that protects against genotoxic stress. Hum. Mol. Genet. (0964-6906) 2004 Gueven N., Becherel OJ., Kijas AW., Chen P., Howe O., Rudolph JH., Gatti R., Date H., Onodera O., Taucher-Scholz G., Lavin MF. Apoptosis - Interactions involving proteins with a function related to apoptosis Chromatin - Epigenetic interactions resulting in chromatin modulation nurig@qimr.edu.au 27-Aug-2008: Contacted by JYOTI. davet human-c3abr Homo sapiens lymphoblastoid cell line c3abr C3ABR ymphoblastoid cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Gueven N., Becherel OJ., Kijas AW., Chen P., Howe O., Rudolph JH., Gatti R., Date H., Onodera O., Taucher-Scholz G., Lavin MF. Apoptosis - Interactions involving proteins with a function related to apoptosis nurig@qimr.edu.au Accepted 2008-SEP-13 by DAVET. 27-Aug-2008: Contacted by JYOTI. Hum. Mol. Genet. (0964-6906) 2004 davet nucl_human Nucleolin NCL Protein C23 protein protein human Homo sapiens Human MVKLAKAGKNQGDPKKMAPPPKEVEEDSEDEEMSEDEEDDSSGEEVVIPQKKGKKAAATSAKKVVVSPTKKVAVATPAKKAAVTPGKKAAATPAKKTVTPAKAVTTPGKKGATPGKALVATPGKKGAAIPAKGAKNGKNAKKEDSDEEEDDDSEEDEEDDEDEDEDEDEIEPAAMKAAAAAPASEDEDDEDDEDDEDDDDDEEDDSEEEAMETTPAKGKKAAKVVPVKAKNVAEDEDEEEDDEDEDDDDDEDDEDDDDEDDEEEEEEEEEEPVKEAPGKRKKEMAKQKAAPEAKKQKVEGTEPTTAFNLFVGNLNFNKSAPELKTGISDVFAKNDLAVVDVRIGMTRKFGYVDFESAEDLEKALELTGLKVFGNEIKLEKPKGKDSKKERDARTLLAKNLPYKVTQDELKEVFEDAAEIRLVSKDGKSKGIAYIEFKTEADAEKTFEEKQGTEIDGRSISLYYTGEKGQNQDYRGGKNSTWSGESKTLVLSNLSYSATEETLQEVFEKATFIKVPQNQNGKSKGYAFIEFASFEDAKEALNSCNKREIEGRAIRLELQGPRGSPNARSQPSKTLFVKGLSEDTTEETLKESFDGSVRARIVTDRETGSSKGFGFVDFNSEEDAKAAKEAMEDGEIDGNKVTLDWAKPKGEGGFGGRGGGRGGFGGRGGGRGGRGGFGGRGRGGFGGRGGFRGGRGGGGDHKPQGKKTKFE DIP protein P19338 original sequence version: 93 DIP protein P19338 original sequence version: 123 C97F6E34E5CA6727 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 aptx_human Aprataxin Forkhead-associated domain histidine triad-like protein APTX AXA1 protein protein human Homo sapiens Human MSNVNLSVSDFWRVMMRVCWLVRQDSRHQRIRLPHLEAVVIGRGPETKITDKKCSRQQVQLKAECNKGYVKVKQVGVNPTSIDSVVIGKDQEVKLQPGQVLHMVNELYPYIVEFEEEAKNPGLETHRKRKRSGNSDSIERDAAQEAEAGTGLEPGSNSGQCSVPLKKGKDAPIKKESLGHWSQGLKISMQDPKMQVYKDEQVVVIKDKYPKARYHWLVLPWTSISSLKAVAREHLELLKHMHTVGEKVIVDFAGSSKLRFRLGYHAIPSMSHVHLHVISQDFDSPCLKNKKHWNSFNTEYFLESQAVIEMVQEAGRVTVRDGMPELLKLPLRCHECQQLLPSIPQLKEHLRKHWTQ 5B338490E35EC8E4 xrcc1_human DNA repair protein XRCC1 XRCC1 X-ray repair cross-complementing protein 1 protein protein human Homo sapiens Human MPEIRLRHVVSCSSQDSTHCAENLLKADTYRKWRAAKAGEKTISVVLQLEKEEQIHSVDIGNDGSAFVEVLVGSSAGGAGEQDYEVLLVTSSFMSPSESRSGSNPNRVRMFGPDKLVRAAAEKRWDRVKIVCSQPYSKDSPFGLSFVRFHSPPDKDEAEAPSQKVTVTKLGQFRVKEEDESANSLRPGALFFSRINKTSPVTASDPAGPSYAAATLQASSAASSASPVSRAIGSTSKPQESPKGKRKLDLNQEEKKTPSKPPAQLSPSVPKRPKLPAPTRTPATAPVPARAQGAVTGKPRGEGTEPRRPRAGPEELGKILQGVVVVLSGFQNPFRSELRDKALELGAKYRPDWTRDSTHLICAFANTPKYSQVLGLGGRIVRKEWVLDCHRMRRRLPSRRYLMAGPGSSSEEDEASHSGGSGDEAPKLPQKQPQTKTKPTQAAGPSSPQKPPTPEETKAASPVLQEDIDIEGVQSEGQDNGAEDSGDTEDELRRVAEQKEHRLPPGQEENGEDPYAGSTDENTDSEEHQEPPDLPVPELPDFFQGKHFFLYGEFPGDERRKLIRYVTAFNGELEDNMSDRVQFVITAQEWDPSFEEALMDNPSLAFVRPRWIYSCNEKQKLLPHQLYGVVPQA DIP protein P18887 original sequence version: 120 DIP protein P18887 original sequence version: 161 30DB3321234DBFC2 parp1_human Poly [ADP-ribose] polymerase 1 PARP1 ADPRT PPOL NAD(+) ADP-ribosyltransferase 1 Poly[ADP-ribose] synthase 1 ADP-ribosyltransferase diphtheria toxin-like 1 DNA ADP-ribosyltransferase PARP1 Protein poly-ADP-ribosyltransferase PARP1 protein protein human Homo sapiens Human MAESSDKLYRVEYAKSGRASCKKCSESIPKDSLRMAIMVQSPMFDGKVPHWYHFSCFWKVGHSIRHPDVEVDGFSELRWDDQQKVKKTAEAGGVTGKGQDGIGSKAEKTLGDFAAEYAKSNRSTCKGCMEKIEKGQVRLSKKMVDPEKPQLGMIDRWYHPGCFVKNREELGFRPEYSASQLKGFSLLATEDKEALKKQLPGVKSEGKRKGDEVDGVDEVAKKKSKKEKDKDSKLEKALKAQNDLIWNIKDELKKVCSTNDLKELLIFNKQQVPSGESAILDRVADGMVFGALLPCEECSGQLVFKSDAYYCTGDVTAWTKCMVKTQTPNRKEWVTPKEFREISYLKKLKVKKQDRIFPPETSASVAATPPPSTASAPAAVNSSASADKPLSNMKILTLGKLSRNKDEVKAMIEKLGGKLTGTANKASLCISTKKEVEKMNKKMEEVKEANIRVVSEDFLQDVSASTKSLQELFLAHILSPWGAEVKAEPVEVVAPRGKSGAALSKKSKGQVKEEGINKSEKRMKLTLKGGAAVDPDSGLEHSAHVLEKGGKVFSATLGLVDIVKGTNSYYKLQLLEDDKENRYWIFRSWGRVGTVIGSNKLEQMPSKEDAIEHFMKLYEEKTGNAWHSKNFTKYPKKFYPLEIDYGQDEEAVKKLTVNPGTKSKLPKPVQDLIKMIFDVESMKKAMVEYEIDLQKMPLGKLSKRQIQAAYSILSEVQQAVSQGSSDSQILDLSNRFYTLIPHDFGMKKPPLLNNADSVQAKVEMLDNLLDIEVAYSLLRGGSDDSSKDPIDVNYEKLKTDIKVVDRDSEEAEIIRKYVKNTHATTHNAYDLEVIDIFKIEREGECQRYKPFKQLHNRRLLWHGSRTTNFAGILSQGLRIAPPEAPVTGYMFGKGIYFADMVSKSANYCHTSQGDPIGLILLGEVALGNMYELKHASHISKLPKGKHSVKGLGKTTPDPSANISLDGVDVPLGTGISSGVNDTSLLYNEYIVYDIAQVNLKYLLKLKFNFKTSLW DIP protein P09874 original sequence version: 189 DIP protein P09874 original sequence version: 183 DIP protein P09874 original sequence version: 166 DIP protein P09874 original sequence version: 117 DIP protein P09874 original sequence version: 143 DIP protein P09874 original sequence version: 136 DIP protein P09874 original sequence version: 137 DIP protein P09874 original sequence version: 147 DIP protein P09874 original sequence version: 193 DIP protein P09874 original sequence version: 202 DIP protein P09874 original sequence version: 171 DIP protein P09874 original sequence version: 153 DIP protein P09874 original sequence version: 175 6A5FC01EB91C046B aptx-xccr1 3 5 unspecified role unspecified role prey prey human-c3abr Homo sapiens lymphoblastoid cell line c3abr C3ABR ymphoblastoid cells 7 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role prey prey human-c3abr Homo sapiens lymphoblastoid cell line c3abr C3ABR ymphoblastoid cells 6 unspecified role unspecified role prey prey human-c3abr Homo sapiens lymphoblastoid cell line c3abr C3ABR ymphoblastoid cells 8 unspecified role unspecified role prey prey human-c3abr Homo sapiens lymphoblastoid cell line c3abr C3ABR ymphoblastoid cells association association 7C The results in Figure 7C show that XRCC1 binds only to the FHA domain of aprataxin. Interestingly, PARP-1 bound to the FHA, NL and HIT domains, and p53 bound to both the FHA and HIT domains. Since aprataxin co-localizes with nucleolin in the nucleoli and since nucleolin is implicated in the genotoxic stress response (27), we determined whether nucleolin also interacted with aprataxin. aptx-parp1 3 9 unspecified role unspecified role prey prey human-c3abr Homo sapiens lymphoblastoid cell line c3abr C3ABR ymphoblastoid cells 7 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 nl region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association 7C The results in Figure 7C show that XRCC1 binds only to the FHA domain of aprataxin. Interestingly, PARP-1 bound to the FHA, NL and HIT domains, and p53 bound to both the FHA and HIT domains. Since aprataxin co-localizes with nucleolin in the nucleoli and since nucleolin is implicated in the genotoxic stress response (27), we determined whether nucleolin also interacted with aprataxin. aptx-p53 3 9 unspecified role unspecified role prey prey human-c3abr Homo sapiens lymphoblastoid cell line c3abr C3ABR ymphoblastoid cells 7 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 hit region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey human-c3abr Homo sapiens lymphoblastoid cell line c3abr C3ABR ymphoblastoid cells association association 7C The results in Figure 7C show that XRCC1 binds only to the FHA domain of aprataxin. Interestingly, PARP-1 bound to the FHA, NL and HIT domains, and p53 bound to both the FHA and HIT domains. Since aprataxin co-localizes with nucleolin in the nucleoli and since nucleolin is implicated in the genotoxic stress response (27), we determined whether nucleolin also interacted with aprataxin. aptx-xccr1-1 4 6 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey association association 7A, 7B Co-immunprecipitation of aprataxin with PARP-1, p53 and XRCC1 Interaction of the P53 with the complex is increased after H2O2 treatment. aptx-parp 4 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey association association 7A Co-immunprecipitation of aprataxin with PARP-1, p53 and XRCC1 aptx-xrcc1-2 4 9 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey association association 7A Co-immunprecipitation of aprataxin with PARP-1, p53 and XRCC1 aptx-ncl 1 5 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component colocalization colocalization 2 we looked for co-localization of aprataxin with the nucleolus-specific marker protein, nucleolin, in stably transfected HeLa cells using anti nucleolin antibody staining (Fig. 2C). When this fluorescence pattern was merged with the EGFP aprataxin signal, co localization of these proteins was observed. aptx-xccr1-2 2 7 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey association association 7A, 7B Co-immunprecipitation of aprataxin with PARP-1, p53 and XRCC1 Treatment of cells with H2O2 increased the incorporation of P53 in this complex. aptx-parp-1 2 9 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey association association 7A Co-immunprecipitation of aprataxin with PARP-1, p53 and XRCC1 aptx-xrcc1-3 2 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey association association 7A Co-immunprecipitation of aprataxin with PARP-1, p53 and XRCC1 aptx-xrcc1-4 1 7 unspecified role unspecified role neutral component neutral component region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role neutral component neutral component colocalization colocalization 9 The interaction was observed in the presence of irradiation with Uranium ions Co-localization of aprataxin and XRCC1 charged particle tracks in chromatin.