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/ Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. J. Biol. Chem. (0021-9258) 2005 imex curation Golubovskaya VM., Finch R., Cance WG. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer cance@surgery.ufl.edu 22-Jan-2008: Contacted by JYOTI. orchard Only protein-protein interactions human-bt474 Homo sapiens breast ductal carcinoma cell line bt474 Human breast ductal carcinoma cell line BT474 Human breast ductal carcinoma cell line established from a solid, invasive ductal carcinoma of the breast obtained from a 60-year-old woman. confocal microscopy confocal microscopy immunostaining immunostaining Golubovskaya VM., Finch R., Cance WG. Apoptosis - Interactions involving proteins with a function related to apoptosis Cells were transfected with GFP-FAK constructs or endogenous FAK was visualized using immunostaining. Endogenous p53 was visualized using immunostaining. cance@surgery.ufl.edu Accepted 2007-DEC-15 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2005 orchard Only protein-protein interactions imex curation Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. J. Biol. Chem. (0021-9258) 2005 imex curation Golubovskaya VM., Finch R., Cance WG. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer cance@surgery.ufl.edu 22-Jan-2008: Contacted by JYOTI. orchard Only protein-protein interactions 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 Golubovskaya VM., Finch R., Cance WG. Apoptosis - Interactions involving proteins with a function related to apoptosis cance@surgery.ufl.edu Accepted 2007-DEC-15 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2005 orchard Only protein-protein interactions imex curation Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. J. Biol. Chem. (0021-9258) 2005 imex curation Golubovskaya VM., Finch R., Cance WG. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer cance@surgery.ufl.edu 22-Jan-2008: Contacted by JYOTI. orchard Only protein-protein interactions 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 Golubovskaya VM., Finch R., Cance WG. Apoptosis - Interactions involving proteins with a function related to apoptosis cance@surgery.ufl.edu Accepted 2007-DEC-15 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2005 orchard Only protein-protein interactions imex curation Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. J. Biol. Chem. (0021-9258) 2005 imex curation Golubovskaya VM., Finch R., Cance WG. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer cance@surgery.ufl.edu 22-Jan-2008: Contacted by JYOTI. orchard Only protein-protein interactions human-hct116 Human colon carcinoma HCT116 cells hct116 Human colon carcinoma line HCT116 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Golubovskaya VM., Finch R., Cance WG. Apoptosis - Interactions involving proteins with a function related to apoptosis cance@surgery.ufl.edu Accepted 2007-DEC-15 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2005 orchard Only protein-protein interactions imex curation Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. J. Biol. Chem. (0021-9258) 2005 imex curation Golubovskaya VM., Finch R., Cance WG. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer cance@surgery.ufl.edu 22-Jan-2008: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro pull down pull down predetermined predetermined participant predetermined Golubovskaya VM., Finch R., Cance WG. Apoptosis - Interactions involving proteins with a function related to apoptosis cance@surgery.ufl.edu Accepted 2007-DEC-15 by ORCHARD. J. Biol. Chem. (0021-9258) 2005 orchard Only protein-protein interactions imex curation Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. J. Biol. Chem. (0021-9258) 2005 imex curation Golubovskaya VM., Finch R., Cance WG. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer cance@surgery.ufl.edu 22-Jan-2008: Contacted by JYOTI. orchard Only protein-protein interactions human-bt474 Homo sapiens breast ductal carcinoma cell line bt474 Human breast ductal carcinoma cell line BT474 Human breast ductal carcinoma cell line established from a solid, invasive ductal carcinoma of the breast obtained from a 60-year-old woman. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Golubovskaya VM., Finch R., Cance WG. Apoptosis - Interactions involving proteins with a function related to apoptosis cance@surgery.ufl.edu Accepted 2007-DEC-15 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2005 orchard Only protein-protein interactions imex curation Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. Direct interaction of the N-terminal domain of focal adhesion kinase with the N-terminal transactivation domain of p53. J. Biol. Chem. (0021-9258) 2005 imex curation Golubovskaya VM., Finch R., Cance WG. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer cance@surgery.ufl.edu 22-Jan-2008: Contacted by JYOTI. orchard Only protein-protein interactions human-cls Homo sapiens fibroblast cell line (GM03318) cls Human fibroblast obtained from patients with Coffin-Lowry Syndrome (CLS). GM03318 (NIGMS Human Genetic Cell Repository) Catalog ID GM03318 (NIGMS Human Genetic Cell Repository) anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Golubovskaya VM., Finch R., Cance WG. Apoptosis - Interactions involving proteins with a function related to apoptosis NHF-1 cells were treated or not treated with doxorubicin which caused significant induction of p53 due to stress. cance@surgery.ufl.edu Accepted 2007-DEC-15 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2005 orchard Only protein-protein interactions imex curation fak1_human Focal adhesion kinase 1 PTK2 FAK FAK1 pp125FAK Protein-tyrosine kinase 2 Focal adhesion kinase-related nonkinase Protein phosphatase 1 regulatory subunit 71 p125FAK protein protein human Homo sapiens Human MAAAYLDPNLNHTPNSSTKTHLGTGMERSPGAMERVLKVFHYFESNSEPTTWASIIRHGDATDVRGIIQKIVDSHKVKHVACYGFRLSHLRSEEVHWLHVDMGVSSVREKYELAHPPEEWKYELRIRYLPKGFLNQFTEDKPTLNFFYQQVKSDYMLEIADQVDQEIALKLGCLEIRRSYWEMRGNALEKKSNYEVLEKDVGLKRFFPKSLLDSVKAKTLRKLIQQTFRQFANLNREESILKFFEILSPVYRFDKECFKCALGSSWIISVELAIGPEEGISYLTDKGCNPTHLADFTQVQTIQYSNSEDKDRKGMLQLKIAGAPEPLTVTAPSLTIAENMADLIDGYCRLVNGTSQSFIIRPQKEGERALPSIPKLANSEKQGMRTHAVSVSETDDYAEIIDEEDTYTMPSTRDYEIQRERIELGRCIGEGQFGDVHQGIYMSPENPALAVAIKTCKNCTSDSVREKFLQEALTMRQFDHPHIVKLIGVITENPVWIIMELCTLGELRSFLQVRKYSLDLASLILYAYQLSTALAYLESKRFVHRDIAARNVLVSSNDCVKLGDFGLSRYMEDSTYYKASKGKLPIKWMAPESINFRRFTSASDVWMFGVCMWEILMHGVKPFQGVKNNDVIGRIENGERLPMPPNCPPTLYSLMTKCWAYDPSRRPRFTELKAQLSTILEEEKAQQEERMRMESRRQATVSWDSGGSDEAPPKPSRPGYPSPRSSEGFYPSPQHMVQTNHYQVSGYPGSHGITAMAGSIYPGQASLLDQTDSWNHRPQEIAMWQPNVEDSTVLDLRGIGQVLPTHLMEERLIRQQQEMEEDQRWLEKEERFLKPDVRLSRGSIDREDGSLQGPIGNQHIYQPVGKPDPAAPPKKPPRPGAPGHLGSLASLSSPADSYNEGVKLQPQEISPPPTANLDRSNDKVYENVTGLVKAVIEMSSKIQPAPPEEYVPMVKEVGLALRTLLATVDETIPLLPASTHREIEMAQKLLNSDLGELINKMKLAQQYVMTSLQQEYKKQMLTAAHALAVDAKNLLDVIDQARLKMLGQTRPH D8A4C15138AB0243 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 ptk2-tp53 5 9 unspecified role unspecified role prey prey 8 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 sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction The p53 was baculovirus-expressed but the cell type was not specified. 1 A, 5 The pull-down assays showed strong binding of p53 with the FAK-NT domain of FAK p73-alpha, a homolog of p53, did not bind GST-FAK, confirming the specificity of this interaction. tp53-ptk2 5 9 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 n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 20 18 direct interaction direct interaction 1 C, 4 We show that full-length FAK binds to GST-p53 PYK-2, a FAK family member, did not bind GST-p53, confirming the specificity of this interaction. tp53-ptk2-1 2 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association Immunoprecipitation of p53 detected FAK in the complex with p53 2 A ptk2-tp53-1 2 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association 2 A, B immunoprecipitation of FAK detected p53 in the complex with FAK Treatment with p53 siRNA reduced the amount of p53 in complex with FAK. ptk2-tp53-2 5 8 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 n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role prey prey 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. direct interaction direct interaction 2 C we detected binding of the p53 protein in 293 cells with the GST-FAK-NT protein, and the binding of p53 with GST-FAK-NT was higher in 293-p53 cells that overexpressed p53 versus 293-vector cells The binding of p53 to GST-FAK-NT was higher in lysates from cells overexpressing p53. tp53-ptk2-2 4 9 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association we performed immunoprecipitation of p53 in colon cancer HCT116 cells, followed by Western blot with FAK antibody (Fig. 2D). We detected FAK protein in the complex with p53 protein in HCT116 cells 2 D tp53-ptk2-3 7 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association we immunoprecipitated p53 in these cells without and with doxorubicin treatment and performed Western blot with FAK antibody (Fig. 2E, lower panel). The FAK-p53 complex was detected in NHF-1 fibroblasts and increased in doxorubicin-treated cells with increased level of p53. 2 E Doxorubicin - Complex formation was enhanced in cells treated with doxorubicin 0.5.microg/ml for 24 h. ptk2-tp53-3 1 9 unspecified role unspecified role neutral component neutral component 8 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 n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain colocalization colocalization 3 A GFP-FAK co-localized with p53 in the nuclear, perinuclear, and cytoplasmic speckles Colocalization took place in nuclear, perinuclear and cytoplasmic speckles. Q05397 transfection ptk2-tp53-4 1 9 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization 3 A Colocalization took place in nuclear, perinuclear and cytoplasmic speckles. Endogenous FAK and p53 clearly co-localized in both cytoplasmic and nuclear areas in BT-474 cells ptk2-tp53-5 6 8 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association We performed immunoprecipitation of FAK from cytoplasmic and nuclear fractions in these cells and detected binding of p53 protein and FAK proteins in both nuclear and cytoplasmic fractions 3 D This complex was detected in both nuclear and cytoplasmic fractions, but was lower in the nucleus as these cells express lower nuclear p53. ptk2-tp53-6 3 8 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association We performed immunoprecipitation of FAK from cytoplasmic and nuclear fractions in these cells and detected binding of p53 protein and FAK proteins in both nuclear and cytoplasmic fractions 3 E This complex was higher in the nuclear fraction in 293 cells as cells express higher levels of nuclear p53.