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/ http://www.ebi.ac.uk/ de_bruijn-2002-3 The cancer-related protein SSX2 interacts with the human homologue of a Ras-like GTPase interactor, RAB3IP, and a novel nuclear protein, SSX2IP. 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 Human testis cDNA library Genes Chromosomes Cancer (1045-2257) 2002 Only protein-protein interactions imex curation de_bruijn-2002-1 The cancer-related protein SSX2 interacts with the human homologue of a Ras-like GTPase interactor, RAB3IP, and a novel nuclear protein, SSX2IP. 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 Human testis cDNA library Genes Chromosomes Cancer (1045-2257) 2002 Only protein-protein interactions imex curation de_bruijn-2002-2 The cancer-related protein SSX2 interacts with the human homologue of a Ras-like GTPase interactor, RAB3IP, and a novel nuclear protein, SSX2IP. 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 Human testis cDNA library Genes Chromosomes Cancer (1045-2257) 2002 Only protein-protein interactions imex curation de_bruijn-2002-4 The cancer-related protein SSX2 interacts with the human homologue of a Ras-like GTPase interactor, RAB3IP, and a novel nuclear protein, SSX2IP. 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 Human testis cDNA library Genes Chromosomes Cancer (1045-2257) 2002 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 q96qf0-2 Rab-3A-interacting protein alpha1 RAB3IP Rabin-3 SSX2-interacting protein RABIN8 protein protein human Homo sapiens Human MANDPLEGFHEVNLASPTSPDLLGVYESGTQEQTTSPSVIYRPHPSALSSVPIQANALDVSELPTQPVYSSPRRLNCAEISSISFHVTDPAPCSTSGVTAGLTKLTTRKDNYNAEREFLQGATITEACDGSDDIFGLSTDSLSRLRSPSVLEVREKGYERLKEELAKAQRELKLKDEECERLSKVRDQLGQELEELTASLFEEAHKMVREANIKQATAEKQLKEAQGKIDVLQAEVAALKTLVLSSSPTSPTQEPLPGGKTPFKKGHTRNKSTSSAMSGSHQDLSVIQPIVKDCKEADLSLYNEFRLWKDEPTMDRTCPFLDKIYQEDIFPCLTFSKSELASAVLEAVENNTLSIEPVGLQPIRFVKASAVECGGPKKCALTGQSKSCKHRIKLGDSSNYYYISPFCRYRITSVCNFFTYIRYIQQGLVKQQDVDQMFWEVMQLRKEMSLAKLGYFKEEL 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 rab3i_human Rab-3A-interacting protein Rabin-3 SSX2-interacting protein RAB3IP RABIN8 protein protein human Homo sapiens Human MGLKKMKGLSYDEAFAMANDPLEGFHEVNLASPTSPDLLGVYESGTQEQTTSPSVIYRPHPSALSSVPIQANALDVSELPTQPVYSSPRRLNCAEISSISFHVTDPAPCSTSGVTAGLTKLTTRKDNYNAEREFLQGATITEACDGSDDIFGLSTDSLSRLRSPSVLEVREKGYERLKEELAKAQRELKLKDEECERLSKVRDQLGQELEELTASLFEEAHKMVREANIKQATAEKQLKEAQGKIDVLQAEVAALKTLVLSSSPTSPTQEPLPGGKTPFKKGHTRNKSTSSAMSGSHQDLSVIQPIVKDCKEADLSLYNEFRLWKDEPTMDRTCPFLDKIYQEDIFPCLTFSKSELASAVLEAVENNTLSIEPVGLQPIRFVKASAVECGGPKKCALTGQSKSCKHRIKLGDSSNYYYISPFCRYRITSVCNFFTYIRYIQQGLVKQQDVDQMFWEVMQLRKEMSLAKLGYFKEEL ssx2-rab3ip-3 Interaction between SSX2 and RAB3IP identified by Y2H 793091 n/a 793095 unspecified role unspecified role bait bait n/a 793096 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 false physical association physical association false false false 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 Interaction between SSX2 and SSX2IP identified by Y2H 793091 n/a 793097 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 false n/a 793095 unspecified role unspecified role bait bait physical association physical association false false false Fig. 3 ssx2-ssx2ip-5 Interaction between SSX2 and SSX2IP identified by Y2H 793091 n/a 793097 unspecified role unspecified role prey prey n/a 793095 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 false physical association physical association false false false Fig. 3A ssx2-rab3ip-5 Interaction between SSX2 and RAB3IP identified by Y2H 793091 n/a 793095 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 false n/a 793096 unspecified role unspecified role prey prey physical association physical association false false false Fig. 3A ssx2-rab3ip-1 Interaction between SSX2 and RAB3IP identified by Y2H 793092 n/a 793095 unspecified role unspecified role bait bait n/a 793098 unspecified role unspecified role prey prey physical association physical association false false false 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 Interaction between SSX2 and SSX2IP identified by Y2H 793092 n/a 793095 unspecified role unspecified role bait bait n/a 793097 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 false physical association physical association false false false ssx2-rab3ip-2 Interaction between SSX2 and RAB3IP identified by pulldown 793093 n/a 793095 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 false region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 793096 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 false in vitro In vitro direct interaction direct interaction false false false Fig. 2A ssx2-ssx2ip-2 Interaction between SSX2 and SSX2IP identified by pulldown 793093 n/a 793097 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 false in vitro In vitro n/a 793095 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 false region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Fig. 2B ssx2-rab3ip-4 Colocalisation of SSX2 and RAB3IP 793094 n/a 793095 unspecified role unspecified role neutral component neutral component n/a 793096 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 false colocalization colocalization false false false Fig. 6 ssx2-ssx2ip-4 Colocalisation of SSX2 and SSX2IP 793094 n/a 793097 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 false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 793095 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig. 8