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/ The cancer-related protein SSX2 interacts with the human homologue of a Ras-like GTPase interactor, RAB3IP, and a novel nuclear protein, SSX2IP. The cancer-related protein SSX2 interacts with the human homologue of a Ras-like GTPase interactor, RAB3IP, and a novel nuclear protein, SSX2IP. Genes Chromosomes Cancer (1045-2257) 2002 imex curation de Bruijn DR., dos Santos NR., Kater-Baats E., Thijssen J., van den Berk L., Stap J., Balemans M., Schepens M., Merkx G., van Kessel AG. Cancer - Interactions investigated in the context of cancer Source of SSX2 taken from PMID:9285793 and subsequent work by the author d.debruin@antrg.azn.nl 09-April-2009: Contacted by JYOTI. Human testis cDNA library davet 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 de Bruijn DR., dos Santos NR., Kater-Baats E., Thijssen J., van den Berk L., Stap J., Balemans M., Schepens M., Merkx G., van Kessel AG. Cancer - Interactions investigated in the context of cancer Source of SSX2 taken from PMID:9285793 and subsequent work by the author d.debruin@antrg.azn.nl Accepted 2009-MAR-9 by DAVET. 09-April-2009: Contacted by JYOTI. Human testis cDNA library Genes Chromosomes Cancer (1045-2257) 2002 davet Only protein-protein interactions imex curation The cancer-related protein SSX2 interacts with the human homologue of a Ras-like GTPase interactor, RAB3IP, and a novel nuclear protein, SSX2IP. The cancer-related protein SSX2 interacts with the human homologue of a Ras-like GTPase interactor, RAB3IP, and a novel nuclear protein, SSX2IP. Genes Chromosomes Cancer (1045-2257) 2002 imex curation de Bruijn DR., dos Santos NR., Kater-Baats E., Thijssen J., van den Berk L., Stap J., Balemans M., Schepens M., Merkx G., van Kessel AG. Cancer - Interactions investigated in the context of cancer Source of SSX2 taken from PMID:9285793 and subsequent work by the author d.debruin@antrg.azn.nl 09-April-2009: Contacted by JYOTI. Human testis cDNA library davet Only protein-protein interactions 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 de Bruijn DR., dos Santos NR., Kater-Baats E., Thijssen J., van den Berk L., Stap J., Balemans M., Schepens M., Merkx G., van Kessel AG. Cancer - Interactions investigated in the context of cancer Fluorescence-tagged proteins observed by direct fluorescence. Source of SSX2 taken from PMID:9285793 and subsequent work by the author d.debruin@antrg.azn.nl Accepted 2009-MAR-9 by DAVET. 09-April-2009: Contacted by JYOTI. Human testis cDNA library Genes Chromosomes Cancer (1045-2257) 2002 davet Only protein-protein interactions imex curation The cancer-related protein SSX2 interacts with the human homologue of a Ras-like GTPase interactor, RAB3IP, and a novel nuclear protein, SSX2IP. The cancer-related protein SSX2 interacts with the human homologue of a Ras-like GTPase interactor, RAB3IP, and a novel nuclear protein, SSX2IP. Genes Chromosomes Cancer (1045-2257) 2002 imex curation de Bruijn DR., dos Santos NR., Kater-Baats E., Thijssen J., van den Berk L., Stap J., Balemans M., Schepens M., Merkx G., van Kessel AG. Cancer - Interactions investigated in the context of cancer Source of SSX2 taken from PMID:9285793 and subsequent work by the author d.debruin@antrg.azn.nl 09-April-2009: Contacted by JYOTI. Human testis cDNA library davet 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 de Bruijn DR., dos Santos NR., Kater-Baats E., Thijssen J., van den Berk L., Stap J., Balemans M., Schepens M., Merkx G., van Kessel AG. Cancer - Interactions investigated in the context of cancer Source of SSX2 taken from PMID:9285793 and subsequent work by the author d.debruin@antrg.azn.nl Accepted 2009-MAR-9 by DAVET. 09-April-2009: Contacted by JYOTI. Human testis cDNA library Genes Chromosomes Cancer (1045-2257) 2002 davet Only protein-protein interactions imex curation The cancer-related protein SSX2 interacts with the human homologue of a Ras-like GTPase interactor, RAB3IP, and a novel nuclear protein, SSX2IP. The cancer-related protein SSX2 interacts with the human homologue of a Ras-like GTPase interactor, RAB3IP, and a novel nuclear protein, SSX2IP. Genes Chromosomes Cancer (1045-2257) 2002 imex curation de Bruijn DR., dos Santos NR., Kater-Baats E., Thijssen J., van den Berk L., Stap J., Balemans M., Schepens M., Merkx G., van Kessel AG. Cancer - Interactions investigated in the context of cancer Source of SSX2 taken from PMID:9285793 and subsequent work by the author d.debruin@antrg.azn.nl 09-April-2009: Contacted by JYOTI. Human testis cDNA library davet Only protein-protein interactions in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra de Bruijn DR., dos Santos NR., Kater-Baats E., Thijssen J., van den Berk L., Stap J., Balemans M., Schepens M., Merkx G., van Kessel AG. Cancer - Interactions investigated in the context of cancer Source of SSX2 taken from PMID:9285793 and subsequent work by the author d.debruin@antrg.azn.nl Accepted 2009-MAR-9 by DAVET. 09-April-2009: Contacted by JYOTI. Human testis cDNA library Genes Chromosomes Cancer (1045-2257) 2002 davet Only protein-protein interactions imex curation ssx2_human Protein SSX2 SSX2 SSX2B SSX2A Synovial sarcoma, X breakpoint 2 Tumor antigen HOM-MEL-40 Cancer/testis antigen 5.2 protein protein human Homo sapiens Human MNGDDAFARRPTVGAQIPEKIQKAFDDIAKYFSKEEWEKMKASEKIFYVYMKRKYEAMTKLGFKATLPPFMCNKRAEDFQGNDLDNDPNRGNQVERPQMTFGRLQGISPKIMPKKPAEEGNDSEEVPEASGPQNDGKELCPPGKPTTSEKIHERSGPKRGEHAWTHRLRERKQLVIYEEISDPEEDDE BF5D18AA5F45B1B1 adip_human Afadin- and alpha-actinin-binding protein SSX2IP KIAA0923 Afadin DIL domain-interacting protein SSX2-interacting protein protein protein human Homo sapiens Human MGDWMTVTDPGLSSESKTISQYTSETKMSPSSLYSQQVLCSSIPLSKNVHSFFSAFCTEDNIEQSISYLDQELTTFGFPSLYEESKGKETKRELNIVAVLNCMNELLVLQRKNLLAQENVETQNLKLGSDMDHLQSCYSKLKEQLETSRREMIGLQERDRQLQCKNRNLHQLLKNEKDEVQKLQNIIASRATQYNHDMKRKEREYNKLKERLHQLVMNKKDKKIAMDILNYVGRADGKRGSWRTGKTEARNEDEMYKILLNDYEYRQKQILMENAELKKVLQQMKKEMISLLSPQKKKPRERVDDSTGTVISDVEEDAGELSRESMWDLSCETVREQLTNSIRKQWRILKSHVEKLDNQVSKVHLEGFNDEDVISRQDHEQETEKLELEIQQCKEMIKTQQQLLQQQLATAYDDDTTSLLRDCYLLEEKERLKEEWSLFKEQKKNFERERRSFTEAAIRLGLERKAFEEERASWLKQQFLNMTTFDHQNSENVKLFSAFSGSSDWDNLIVHSRQPQKKPHSVSNGSPVCMSKLTKSLPASPSTSDFCQTRSCISEHSSINVLNITAEEIKPNQVGGECTNQKWSVASRPGSQEGCYSGCSLSYTNSHVEKDDLP 427903EF86A6FE31 q96qf0-2 Rab-3A-interacting protein alpha1 RAB3IP Rabin-3 SSX2-interacting protein RABIN8 protein protein human Homo sapiens Human MANDPLEGFHEVNLASPTSPDLLGVYESGTQEQTTSPSVIYRPHPSALSSVPIQANALDVSELPTQPVYSSPRRLNCAEISSISFHVTDPAPCSTSGVTAGLTKLTTRKDNYNAEREFLQGATITEACDGSDDIFGLSTDSLSRLRSPSVLEVREKGYERLKEELAKAQRELKLKDEECERLSKVRDQLGQELEELTASLFEEAHKMVREANIKQATAEKQLKEAQGKIDVLQAEVAALKTLVLSSSPTSPTQEPLPGGKTPFKKGHTRNKSTSSAMSGSHQDLSVIQPIVKDCKEADLSLYNEFRLWKDEPTMDRTCPFLDKIYQEDIFPCLTFSKSELASAVLEAVENNTLSIEPVGLQPIRFVKASAVECGGPKKCALTGQSKSCKHRIKLGDSSNYYYISPFCRYRITSVCNFFTYIRYIQQGLVKQQDVDQMFWEVMQLRKEMSLAKLGYFKEEL DIP protein Q96QF0-2 original sequence version: 71 DIP protein Q96QF0-2 original sequence version: 116 ED659E364B5C75FA rab3i_human Rab-3A-interacting protein Rabin-3 SSX2-interacting protein RAB3IP RABIN8 protein protein human Homo sapiens Human MGLKKMKGLSYDEAFAMANDPLEGFHEVNLASPTSPDLLGVYESGTQEQTTSPSVIYRPHPSALSSVPIQANALDVSELPTQPVYSSPRRLNCAEISSISFHVTDPAPCSTSGVTAGLTKLTTRKDNYNAEREFLQGATITEACDGSDDIFGLSTDSLSRLRSPSVLEVREKGYERLKEELAKAQRELKLKDEECERLSKVRDQLGQELEELTASLFEEAHKMVREANIKQATAEKQLKEAQGKIDVLQAEVAALKTLVLSSSPTSPTQEPLPGGKTPFKKGHTRNKSTSSAMSGSHQDLSVIQPIVKDCKEADLSLYNEFRLWKDEPTMDRTCPFLDKIYQEDIFPCLTFSKSELASAVLEAVENNTLSIEPVGLQPIRFVKASAVECGGPKKCALTGQSKSCKHRIKLGDSSNYYYISPFCRYRITSVCNFFTYIRYIQQGLVKQQDVDQMFWEVMQLRKEMSLAKLGYFKEEL DIP protein Q96QF0 original sequence version: 71 DIP protein Q96QF0 original sequence version: 112 DD77781823038F15 ssx2-rab3ip-1 3 5 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association The author identified RAB3IP identified isoforms alpha and gamma as interacting with SSX2 but did not further differentiate these to alpha/gamma 1 or 2. ssx2-ssx2ip-1 3 5 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind full dna sequence full identification by DNA sequencing full dna sequence certain certain sequence position certain certain certain sequence position certain physical association physical association The SSX2IP cDNA fragment initially identified by yeast two-hybrid screening corresponded to the C-terminal end of the protein, encompassing amino acids 384 to 614 ssx2-rab3ip-2 4 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 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) 7 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction GST pull-down assays showing that the RAB3IP and SSX2IP proteins interact in vitro with SSX2 Fig. 2A ssx2-ssx2ip-2 4 6 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 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 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 GST pull-down assays showing that the RAB3IP and SSX2IP proteins interact in vitro with SSX2 Fig. 2B ssx2-rab3ip-3 1 5 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey 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 These results suggest that putative SSX2-interacting domains are either interrupted or absent in all three deletion constructs or, alternatively, that both N-terminal (from position 79) and C-terminal (up to position 304) sequences are required for interaction with SSX2. The binding site shown is one interpretation of the results, alternatively, the putative SSX2-interacting domains are either interrupted or absent in all three deletion constructs used in the exeriment. Fig. 3 ssx2-ssx2ip-3 1 6 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind full dna sequence full identification by DNA sequencing full dna sequence certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role bait bait physical association physical association The SSX2IP cDNA fragment initially identified by yeast two-hybrid screening corresponded to the C-terminal end of the protein, encompassing amino acids 384 to 614 Fig. 3 ssx2-rab3ip-4 2 5 unspecified role unspecified role neutral component neutral component 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 colocalization colocalization Subcellular localization of the EGFP-RAB3IP and SSX2 proteins in transfected HeLa cells. Fig. 6 ssx2-ssx2ip-4 2 6 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role neutral component neutral component colocalization colocalization Subcellular localization of SSX2IP and SSX2 protein fragments in transfected HeLa cells. Fig. 8 ssx2-ssx2ip-5 1 6 unspecified role unspecified role prey prey 5 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 Thus, both RAB3IP and SSX2IP interact with an N-terminal domain of SSX2, with RAB3IP requiring amino acids 25 to 80 and SSX2IP requiring amino acids 1 to 80. Fig. 3A ssx2-rab3ip-5 1 5 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 7 unspecified role unspecified role prey prey physical association physical association Thus, both RAB3IP and SSX2IP interact with an N-terminal domain of SSX2, with RAB3IP requiring amino acids 25 to 80 and SSX2IP requiring amino acids 1 to 80. Fig. 3A