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/ Arf and Rho GAP adapter protein ARAP1 participates in the mobilization of TRAIL-R1/DR4 to the plasma membrane. Arf and Rho GAP adapter protein ARAP1 participates in the mobilization of TRAIL-R1/DR4 to the plasma membrane. Apoptosis (1360-8185) 2007 imex curation Simova S., Klima M., Cermak L., Sourkova V., Andera L. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Supp. Fig 2B could not be opened for curation as the available file appeared to be corrupted. The positive control was not cuated as the species of origin information was not present in the paper and for a control interaction it was not deemed time efficient to write to the authors. From comparing the authors given range for the intracellular part of DR4 (aa 246-445), it is clear that they have not included the signal peptide of 23 aa that is included in the UniProtKB entry (O00220) for DR4. Thus the range has been altered by 23 to compensate for this discrepancy. andera@img.cas.cz. 16-Dec-2008: Contacted by JYOTI. HA-tagged human cDNA libraries (leukocyte/bone marrow) were used. Only protein-protein interactions human-nctc Homo sapiens skin epithelial-like cells nctc Human skin epithelial-like cells confocal microscopy confocal microscopy immunostaining immunostaining Simova S., Klima M., Cermak L., Sourkova V., Andera L. Apoptosis - Interactions involving proteins with a function related to apoptosis Supp. Fig 2B could not be opened for curation as the available file appeared to be corrupted. The positive control was not cuated as the species of origin information was not present in the paper and for a control interaction it was not deemed time efficient to write to the authors. From comparing the authors given range for the intracellular part of DR4 (aa 246-445), it is clear that they have not included the signal peptide of 23 aa that is included in the UniProtKB entry (O00220) for DR4. Thus the range has been altered by 23 to compensate for this discrepancy. From comparing the authors given range for the intracellular part of DR4 (aa 246-445), it is clear that they have not included the signal peptide of 23 aa that is included in the UniProtKB entry (O00220) for DR4. Thus the range has been altered by 23 to compensate for this discrepancy. andera@img.cas.cz. Accepted 2008-Not a month!-8 by ORCHARD. 16-Dec-2008: Contacted by JYOTI. HA-tagged human cDNA libraries (leukocyte/bone marrow) were used. Apoptosis (1360-8185) 2007 Only protein-protein interactions imex curation Arf and Rho GAP adapter protein ARAP1 participates in the mobilization of TRAIL-R1/DR4 to the plasma membrane. Arf and Rho GAP adapter protein ARAP1 participates in the mobilization of TRAIL-R1/DR4 to the plasma membrane. Apoptosis (1360-8185) 2007 imex curation Simova S., Klima M., Cermak L., Sourkova V., Andera L. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Supp. Fig 2B could not be opened for curation as the available file appeared to be corrupted. The positive control was not cuated as the species of origin information was not present in the paper and for a control interaction it was not deemed time efficient to write to the authors. From comparing the authors given range for the intracellular part of DR4 (aa 246-445), it is clear that they have not included the signal peptide of 23 aa that is included in the UniProtKB entry (O00220) for DR4. Thus the range has been altered by 23 to compensate for this discrepancy. andera@img.cas.cz. 16-Dec-2008: Contacted by JYOTI. HA-tagged human cDNA libraries (leukocyte/bone marrow) were used. Only protein-protein interactions 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 Simova S., Klima M., Cermak L., Sourkova V., Andera L. Apoptosis - Interactions involving proteins with a function related to apoptosis Supp. Fig 2B could not be opened for curation as the available file appeared to be corrupted. The positive control was not cuated as the species of origin information was not present in the paper and for a control interaction it was not deemed time efficient to write to the authors. From comparing the authors given range for the intracellular part of DR4 (aa 246-445), it is clear that they have not included the signal peptide of 23 aa that is included in the UniProtKB entry (O00220) for DR4. Thus the range has been altered by 23 to compensate for this discrepancy. From comparing the authors given range for the intracellular part of DR4 (aa 246-445), it is clear that they have not included the signal peptide of 23 aa that is included in the UniProtKB entry (O00220) for DR4. Thus the range has been altered by 23 to compensate for this discrepancy. andera@img.cas.cz. Accepted 2008-Not a month!-8 by ORCHARD. 16-Dec-2008: Contacted by JYOTI. HA-tagged human cDNA libraries (leukocyte/bone marrow) were used. Apoptosis (1360-8185) 2007 Only protein-protein interactions imex curation Arf and Rho GAP adapter protein ARAP1 participates in the mobilization of TRAIL-R1/DR4 to the plasma membrane. Arf and Rho GAP adapter protein ARAP1 participates in the mobilization of TRAIL-R1/DR4 to the plasma membrane. Apoptosis (1360-8185) 2007 imex curation Simova S., Klima M., Cermak L., Sourkova V., Andera L. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Supp. Fig 2B could not be opened for curation as the available file appeared to be corrupted. The positive control was not cuated as the species of origin information was not present in the paper and for a control interaction it was not deemed time efficient to write to the authors. From comparing the authors given range for the intracellular part of DR4 (aa 246-445), it is clear that they have not included the signal peptide of 23 aa that is included in the UniProtKB entry (O00220) for DR4. Thus the range has been altered by 23 to compensate for this discrepancy. andera@img.cas.cz. 16-Dec-2008: Contacted by JYOTI. HA-tagged human cDNA libraries (leukocyte/bone marrow) were used. 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 bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Simova S., Klima M., Cermak L., Sourkova V., Andera L. Apoptosis - Interactions involving proteins with a function related to apoptosis Supp. Fig 2B could not be opened for curation as the available file appeared to be corrupted. The positive control was not cuated as the species of origin information was not present in the paper and for a control interaction it was not deemed time efficient to write to the authors. From comparing the authors given range for the intracellular part of DR4 (aa 246-445), it is clear that they have not included the signal peptide of 23 aa that is included in the UniProtKB entry (O00220) for DR4. Thus the range has been altered by 23 to compensate for this discrepancy. andera@img.cas.cz. Accepted 2008-Not a month!-8 by ORCHARD. 16-Dec-2008: Contacted by JYOTI. HA-tagged human cDNA libraries (leukocyte/bone marrow) were used. Apoptosis (1360-8185) 2007 Only protein-protein interactions imex curation Arf and Rho GAP adapter protein ARAP1 participates in the mobilization of TRAIL-R1/DR4 to the plasma membrane. Arf and Rho GAP adapter protein ARAP1 participates in the mobilization of TRAIL-R1/DR4 to the plasma membrane. Apoptosis (1360-8185) 2007 imex curation Simova S., Klima M., Cermak L., Sourkova V., Andera L. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Supp. Fig 2B could not be opened for curation as the available file appeared to be corrupted. The positive control was not cuated as the species of origin information was not present in the paper and for a control interaction it was not deemed time efficient to write to the authors. From comparing the authors given range for the intracellular part of DR4 (aa 246-445), it is clear that they have not included the signal peptide of 23 aa that is included in the UniProtKB entry (O00220) for DR4. Thus the range has been altered by 23 to compensate for this discrepancy. andera@img.cas.cz. 16-Dec-2008: Contacted by JYOTI. HA-tagged human cDNA libraries (leukocyte/bone marrow) were used. Only protein-protein interactions 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 Simova S., Klima M., Cermak L., Sourkova V., Andera L. Apoptosis - Interactions involving proteins with a function related to apoptosis Supp. Fig 2B could not be opened for curation as the available file appeared to be corrupted. The positive control was not cuated as the species of origin information was not present in the paper and for a control interaction it was not deemed time efficient to write to the authors. From comparing the authors given range for the intracellular part of DR4 (aa 246-445), it is clear that they have not included the signal peptide of 23 aa that is included in the UniProtKB entry (O00220) for DR4. Thus the range has been altered by 23 to compensate for this discrepancy. From comparing the authors given range for the intracellular part of DR4 (aa 246-445), it is clear that they have not included the signal peptide of 23 aa that is included in the UniProtKB entry (O00220) for DR4. Thus the range has been altered by 23 to compensate for this discrepancy. andera@img.cas.cz. Accepted 2008-Not a month!-8 by ORCHARD. 16-Dec-2008: Contacted by JYOTI. HA-tagged human cDNA libraries (leukocyte/bone marrow) were used. Apoptosis (1360-8185) 2007 Only protein-protein interactions imex curation arap1_human Arf-GAP with Rho-GAP domain, ANK repeat and PH domain-containing protein 1 ARAP1 CENTD2 Centaurin-delta-2 KIAA0782 protein protein human Homo sapiens Human MAEAGDAALSVAEWLRALHLEQYTGLFEQHGLVWATECQGLSDTRLMDMGMLLPGHRRRILAGLLRAHTSPAPAPRPTPRPVPMKRHIFRSPPVPATPPEPLPTTTEDEGLPAAPPIPPRRSCLPPTCFTTPSTAAPDPVLPPLPAKRHLAELSVPPVPPRTGPPRLLVSLPTKEEESLLPSLSSPPQPQSEEPLSTLPQGPPQPPSPPPCPPEIPPKPVRLFPEFDDSDYDEVPEEGPGAPARVMTKKEEPPPSRVPRAVRVASLLSEGEELSGDDQGDEEEDDHAYEGVPNGGWHTSSLSLSLPSTIAAPHPMDGPPGGSTPVTPVIKAGWLDKNPPQGSYIYQKRWVRLDTDHLRYFDSNKDAYSKRFISVACISHVAAIGDQKFEVITNNRTFAFRAESDVERKEWMQALQQAMAEQRARARLSSAYLLGVPGSEQPDRAGSLELRGFKNKLYVAVVGDKVQLYKNLEEYHLGIGITFIDMSVGNVKEVDRRSFDLTTPYRIFSFSADSELEKEQWLEAMQGAIAEALSTSEVAERIWAAAPNRFCADCGAPQPDWASINLCVVICKRCAGEHRGLGAGVSKVRSLKMDRKVWTETLIELFLQLGNGAGNRFWAANVPPSEALQPSSSPSTRRCHLEAKYREGKYRRYHPLFGNQEELDKALCAAVTTTDLAETQALLGCGAGINCFSGDPEAPTPLALAEQAGQTLQMEFLRNNRTTEVPRLDSMKPLEKHYSVVLPTVSHSGFLYKTASAGKLLQDRRAREEFSRRWCVLGDGVLSYFENERAVTPNGEIRASEIVCLAVPPPDTHGFEHTFEVYTEGERLYLFGLESAEQAHEWVKCIAKAFVPPLAEDLLARDFERLGRLPYKAGLSLQRAQEGWFSLSGSELRAVFPEGPCEEPLQLRKLQELSIQGDSENQVLVLVERRRTLYIQGERRLDFMGWLGAIQKAAASMGDTLSEQQLGDSDIPVIVYRCVDYITQCGLTSEGIYRKCGQTSKTQRLLESLRQDARSVHLKEGEQHVDDVSSALKRFLRDLPDGLFTRAQRLTWLEASEIEDEEEKVSRYRELLVRLPPVNRATVKALISHLYCVQCFSDTNQMNVHNLAIVFGPTLFQTDGQDYKAGRVVEDLINHYVVVFSVDEEELRKQREEITAIVKMRVAGTASGTQHAGDFICTVYLEEKKAETEQHIKVPASMTAEELTLEILDRRNVGIREKDYWTCFEVNEREEAERPLHFAEKVLPILHGLGTDSHLVVKKHQAMEAMLLYLASRVGDTKHGMMKFREDRSLLGLGLPSGGFHDRYFILNSSCLRLYKEVRSQRPWSGAPETSHRPEKEWPIKSLKVYLGVKKKLRPPTCWGFTVVHETEKHEKQQWYLCCDTQMELREWFATFLFVQHDGLVWPSEPSRVSRAVPEVRLGSVSLIPLRGSENEMRRSVAAFTADPLSLLRNV A4B6CCC28EC4CD0D tr10b_human Tumor necrosis factor receptor superfamily member 10B Death receptor 5 TNF-related apoptosis-inducing ligand receptor 2 UNQ160/PRO186 TNFRSF10B DR5 KILLER TRAILR2 TRICK2 ZTNFR9 protein protein human Homo sapiens Human MEQRGQNAPAASGARKRHGPGPREARGARPGPRVPKTLVLVVAAVLLLVSAESALITQQDLAPQQRAAPQQKRSSPSEGLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRCDSGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSDIECVHKESGTKHSGEVPAVEETVTSSPGTPASPCSLSGIIIGVTVAAVVLIVAVFVCKSLLWKKVLPYLKGICSGGGGDPERVDRSSQRPGAEDNVLNEIVSILQPTQVPEQEMEVQEPAEPTGVNMLSPGESEHLLEPAEAERSQRRRLLVPANEGDPTETLRQCFDDFADLVPFDSWEPLMRKLGLMDNEIKVAKAEAAGHRDTLYTMLIKWVNKTGRDASVHTLLDALETLGERLAKQKIEDHLLSSGKFMYLEGNADSAMS 60358EAF2A835870 tr10a_human Tumor necrosis factor receptor superfamily member 10A Death receptor 4 TNF-related apoptosis-inducing ligand receptor 1 TNFRSF10A APO2 DR4 TRAILR1 protein protein human Homo sapiens Human MAPPPARVHLGAFLAVTPNPGSAASGTEAAAATPSKVWGSSAGRIEPRGGGRGALPTSMGQHGPSARARAGRAPGPRPAREASPRLRVHKTFKFVVVGVLLQVVPSSAATIKLHDQSIGTQQWEHSPLGELCPPGSHRSEHPGACNRCTEGVGYTNASNNLFACLPCTACKSDEEERSPCTTTRNTACQCKPGTFRNDNSAEMCRKCSRGCPRGMVKVKDCTPWSDIECVHKESGNGHNIWVILVVTLVVPLLLVAVLIVCCCIGSGCGGDPKCMDRVCFWRLGLLRGPGAEDNAHNEILSNADSLSTFVSEQQMESQEPADLTGVTVQSPGEAQCLLGPAEAEGSQRRRLLVPANGADPTETLMLFFDKFANIVPFDSWDQLMRQLDLTKNEIDVVRAGTAGPGDALYAMLMKWVNKTGRNASIHTLLDALERMEERHAREKIQDLLVDSGKFIYLEDGTGSAVSLE 7E96619D0BDC0CD4 tnfrsf10a-centd2 4 5 unspecified role unspecified role prey prey c-terminal region 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 intracellular region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 1 A In order to search for new proteins that interact with TRAIL-R1/DR4 and could affect its signaling, we performed a yeast two-hybrid (Y2H) screening of several cDNA expression libraries using the intracellular part of DR4 (ICP, amino acids 246-445) as a bait. Two fusion proteins specifically interacting with DR4(ICP) contained a C-terminal part of ARAP1 (centaurin d2), an adapter protein with multiple domains and Arf and Rho GAP activities The CENTD2 protein is an isoform lacking 11 amino acids near to the C-terminus but this could not be unambiguously mapped to a single UniprotKB isoform tnfrsf10a-centd2-1 2 5 unspecified role unspecified role prey prey c-terminal region 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 intracellular region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 17 15 physical association physical association 1 B Interaction analysis in yeasts uncovered that the presence of exon 30 in the ARAP1 C-terminus abolished its interaction with the intracellular part of DR4 The presence of exon 30 in the ARAP1 C-terminus abolished its interaction with the intracellular part of DR4. tnfrsf10a-centd2-2 3 5 unspecified role unspecified role prey prey c-terminal region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role bait bait c-terminal required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain O00220:p.[Phe368Pro;Asp369Leu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain F P certain certain sequence position certain certain certain sequence position certain D L Wed May 02 19:31:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe368pro,asp369leu' Tue Jun 30 19:03:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Phe368Pro;Asp369Leu]' physical association physical association 2 A, B Myc-tagged ARAP1-CDexon30 co-immunoprecipitated with DR4 Full-length ARAP1 Delta exon 30 co-immunoprecipitated with DR4, but less efficiently than its C-terminal part. In the DR4 death domain, phe368 asp369 are conserved when compared to DR5. Their substitution with the corresponding amino acids from TNFR1 (proline and leucine) in mutant DR4-FD/PL led to the severe suppression of ARAP1-CDexon30 co-immunoprecipitation with DR4. O00220; Q96P48-3 transfection tnfrsf10b-centd2 3 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 6 unspecified role unspecified role bait bait physical association physical association 2 B Myc-tagged ARAP1-CDexon30 co-immunoprecipitated with DR4 and, to lesser extent, DR5 Myc-tagged ARAP1-CDexon30 co-immunoprecipitated with DR5 but to a lesser extent than with DR4. centd2-tnfrsf10a 1 5 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component colocalization colocalization 3 A In untreated cells transfected DR4 was localized at the membrane and in the ER/Golgi compartment surrounding the nuclei. ARAP1 was positioned in the ER/Golgi compartment in agreement with previously published data, but a significant fraction of ARAP1 was also localized at the cellular membrane. At both sites ARAP1 apparently co-localized with DR4 Upon TRAIL treatment, both ARAP1 and DR4 co-localized in early endosomes