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/ SAFB2, a new scaffold attachment factor homolog and estrogen receptor corepressor. SAFB2, a new scaffold attachment factor homolog and estrogen receptor corepressor. J. Biol. Chem. (0021-9258) 2003 imex curation Townson SM., Dobrzycka KM., Lee AV., Air M., Deng W., Kang K., Jiang S., Kioka N., Michaelis K., Oesterreich S. Cancer - Interactions investigated in the context of cancer Source of ER taken from PMID: 9393950 as described in the text steffio@breastcenter.tmc.edu davet Only protein-protein interactions human-saos-2 Homo sapiens SAOS-2 cell line established from primary osteogenic sarcoma saos-2 Homo sapiens SAOS-2 cell line established from primary osteogenic sarcoma confocal microscopy confocal microscopy immunostaining immunostaining Townson SM., Dobrzycka KM., Lee AV., Air M., Deng W., Kang K., Jiang S., Kioka N., Michaelis K., Oesterreich S. Cancer - Interactions investigated in the context of cancer Fluorescently tagged proteins detected by direct fluorescence. Source of ER taken from PMID: 9393950 as described in the text steffio@breastcenter.tmc.edu Accepted 2009-FEB-29 by DAVET. J. Biol. Chem. (0021-9258) 2003 davet Only protein-protein interactions imex curation SAFB2, a new scaffold attachment factor homolog and estrogen receptor corepressor. SAFB2, a new scaffold attachment factor homolog and estrogen receptor corepressor. J. Biol. Chem. (0021-9258) 2003 imex curation Townson SM., Dobrzycka KM., Lee AV., Air M., Deng W., Kang K., Jiang S., Kioka N., Michaelis K., Oesterreich S. Cancer - Interactions investigated in the context of cancer Source of ER taken from PMID: 9393950 as described in the text steffio@breastcenter.tmc.edu 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 Townson SM., Dobrzycka KM., Lee AV., Air M., Deng W., Kang K., Jiang S., Kioka N., Michaelis K., Oesterreich S. Cancer - Interactions investigated in the context of cancer Source of ER taken from PMID: 9393950 as described in the text steffio@breastcenter.tmc.edu Accepted 2009-FEB-29 by DAVET. J. Biol. Chem. (0021-9258) 2003 davet Only protein-protein interactions imex curation SAFB2, a new scaffold attachment factor homolog and estrogen receptor corepressor. SAFB2, a new scaffold attachment factor homolog and estrogen receptor corepressor. J. Biol. Chem. (0021-9258) 2003 imex curation Townson SM., Dobrzycka KM., Lee AV., Air M., Deng W., Kang K., Jiang S., Kioka N., Michaelis K., Oesterreich S. Cancer - Interactions investigated in the context of cancer Source of ER taken from PMID: 9393950 as described in the text steffio@breastcenter.tmc.edu davet Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Townson SM., Dobrzycka KM., Lee AV., Air M., Deng W., Kang K., Jiang S., Kioka N., Michaelis K., Oesterreich S. Cancer - Interactions investigated in the context of cancer Source of ER taken from PMID: 9393950 as described in the text steffio@breastcenter.tmc.edu Accepted 2009-FEB-29 by DAVET. J. Biol. Chem. (0021-9258) 2003 davet Only protein-protein interactions imex curation SAFB2, a new scaffold attachment factor homolog and estrogen receptor corepressor. SAFB2, a new scaffold attachment factor homolog and estrogen receptor corepressor. J. Biol. Chem. (0021-9258) 2003 imex curation Townson SM., Dobrzycka KM., Lee AV., Air M., Deng W., Kang K., Jiang S., Kioka N., Michaelis K., Oesterreich S. Cancer - Interactions investigated in the context of cancer Source of ER taken from PMID: 9393950 as described in the text steffio@breastcenter.tmc.edu davet Only protein-protein interactions chlae-cv-1 Monkey African Green kidney fibroblast Grivet cv-1 Monkey African Green kidney fibroblast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Townson SM., Dobrzycka KM., Lee AV., Air M., Deng W., Kang K., Jiang S., Kioka N., Michaelis K., Oesterreich S. Cancer - Interactions investigated in the context of cancer Source of ER taken from PMID: 9393950 as described in the text steffio@breastcenter.tmc.edu Accepted 2009-FEB-29 by DAVET. J. Biol. Chem. (0021-9258) 2003 davet Only protein-protein interactions imex curation SAFB2, a new scaffold attachment factor homolog and estrogen receptor corepressor. SAFB2, a new scaffold attachment factor homolog and estrogen receptor corepressor. J. Biol. Chem. (0021-9258) 2003 imex curation Townson SM., Dobrzycka KM., Lee AV., Air M., Deng W., Kang K., Jiang S., Kioka N., Michaelis K., Oesterreich S. Cancer - Interactions investigated in the context of cancer Source of ER taken from PMID: 9393950 as described in the text steffio@breastcenter.tmc.edu davet Only protein-protein interactions human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Townson SM., Dobrzycka KM., Lee AV., Air M., Deng W., Kang K., Jiang S., Kioka N., Michaelis K., Oesterreich S. Cancer - Interactions investigated in the context of cancer Source of ER taken from PMID: 9393950 as described in the text steffio@breastcenter.tmc.edu Accepted 2009-FEB-29 by DAVET. J. Biol. Chem. (0021-9258) 2003 davet Only protein-protein interactions imex curation SAFB2, a new scaffold attachment factor homolog and estrogen receptor corepressor. SAFB2, a new scaffold attachment factor homolog and estrogen receptor corepressor. J. Biol. Chem. (0021-9258) 2003 imex curation Townson SM., Dobrzycka KM., Lee AV., Air M., Deng W., Kang K., Jiang S., Kioka N., Michaelis K., Oesterreich S. Cancer - Interactions investigated in the context of cancer Source of ER taken from PMID: 9393950 as described in the text steffio@breastcenter.tmc.edu davet Only protein-protein interactions human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Townson SM., Dobrzycka KM., Lee AV., Air M., Deng W., Kang K., Jiang S., Kioka N., Michaelis K., Oesterreich S. Cancer - Interactions investigated in the context of cancer Source of ER taken from PMID: 9393950 as described in the text steffio@breastcenter.tmc.edu Accepted 2009-FEB-29 by DAVET. J. Biol. Chem. (0021-9258) 2003 davet Only protein-protein interactions imex curation esr1_human Estrogen receptor ESR1 ESR NR3A1 Estradiol receptor ER-alpha Nuclear receptor subfamily 3 group A member 1 protein protein human Homo sapiens Human MTMTLHTKASGMALLHQIQGNELEPLNRPQLKIPLERPLGEVYLDSSKPAVYNYPEGAAYEFNAAAAANAQVYGQTGLPYGPGSEAAAFGSNGLGGFPPLNSVSPSPLMLLHPPPQLSPFLQPHGQQVPYYLENEPSGYTVREAGPPAFYRPNSDNRRQGGRERLASTNDKGSMAMESAKETRYCAVCNDYASGYHYGVWSCEGCKAFFKRSIQGHNDYMCPATNQCTIDKNRRKSCQACRLRKCYEVGMMKGGIRKDRRGGRMLKHKRQRDDGEGRGEVGSAGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSLQKYYITGEAEGFPATV DIP protein P03372 original sequence version: 214 DIP protein P03372 original sequence version: 139 DIP protein P03372 original sequence version: 167 5455C57AB0CCCAA7 safb1_human Scaffold attachment factor B1 SAFB HAP HET SAFB1 HSP27 estrogen response element-TATA box-binding protein protein protein human Homo sapiens Human MAETLSGLGDSGAAGAAALSSASSETGTRRLSDLRVIDLRAELRKRNVDSSGNKSVLMERLKKAIEDEGGNPDEIEITSEGNKKTSKRSSKGRKPEEEGVEDNGLEENSGDGQEDVETSLENLQDIDIMDISVLDEAEIDNGSVADCVEDDDADNLQESLSDSRELVEGEMKELPEQLQEHAIEDKETINNLDTSSSDFTILQEIEEPSLEPENEKILDILGETCKSEPVKEESSELEQPFAQDTSSVGPDRKLAEEEDLFDSAHPEEGDLDLASESTAHAQSSKADSLLAVVKREPAEQPGDGERTDCEPVGLEPAVEQSSAASELAEASSEELAEAPTEAPSPEARDSKEDGRKFDFDACNEVPPAPKESSTSEGADQKMSSPEDDSDTKRLSKEEKGRSSCGRNFWVSGLSSTTRATDLKNLFSKYGKVVGAKVVTNARSPGARCYGFVTMSTAEEATKCINHLHKTELHGKMISVEKAKNEPVGKKTSDKRDSDGKKEKSSNSDRSTNLKRDDKCDRKDDAKKGDDGSGEKSKDQDDQKPGPSERSRATKSGSRGTERTVVMDKSKGVPVISVKTSGSKERASKSQDRKSASREKRSVVSFDKVKEPRKSRDSESHSRVRERSEREQRMQAQWEREERERLEIARERLAFQRQRLERERMERERLERERMHVEHERRREQERIHREREELRRQQELRYEQERRPAVRRPYDLDRRDDAYWPEAKRAALDERYHSDFNRQDRFHDFDHRDRGRYPDHSVDRREGSRSMMGEREGQHYPERHGGPERHGRDSRDGWGGYGSDKRMSEGRGLPPPPRRDWGDHGRREDDRSWQGTADGGMMDRDHKRWQGGERSMSGHSGPGHMMNRGGMSGRGSFAPGGASRGHPIPHGGMQGGFGGQSRGSRPSDARFTRRY E865940BC7783C4A o60504-2 Vinexin SORBS3 SCAM1 SH3-containing adapter molecule 1 Sorbin and SH3 domain-containing protein 3 protein protein human Homo sapiens Human MADGGSPFLGRRDFVYPSSTRDPSASNGGGSPARREEKKRKAARLKFDFQAQSPKELTLQKGDIVYIHKEVDKNWLEGEHHGRLGIFPANYVEVLPADEIPKPIKPPTYQVLEYGEAVAQYTFKGDLEVELSFRKGEHICLIRKVNENWYEGRITGTGRQGIFPASYVQVSREPRLRLCDDGPQLPTSPRLTAAARSARHPSSPSALRSPADPIDLGGQTSPRRTGFSFPTQEPRPQTQNLGTPGPALSHSRGPSHPLDLGTSSPNTSQIHWTPYRAMYQYRPQNEDELELREGDRVDVMQQCDDGWFVGVSRRTQKFGTFPGNYVAPV Contains a phosphoserine at position 6. Initiator Met-1 is removed. Contains a N-acetylalanine at position 2. 852B1FDC07859AC2 o60504-1 Vinexin SORBS3 SCAM1 SH3-containing adapter molecule 1 Sorbin and SH3 domain-containing protein 3 protein protein human Homo sapiens Human MQGPPRSLRAGLSLDDFIPGHLQSHIGSSSRGTRVPVIRNGGSNTLNFQFHDPAPRTVCNGGYTPRRDASQHPDPAWYQTWPGPGSKPSASTKIPASQHTQNWSATWTKDSKRRDKRWVKYEGIGPVDESGMPIAPRSSVDRPRDWYRRMFQQIHRKMPDLQLDWTFEEPPRDPRHLGAQQRPAHRPGPATSSSGRSWDHSEELPRSTFNYRPGAFSTVLQPSNQVLRRREKVDNVWTEESWNQFLQELETGQRPKKPLVDDPGEKPSQPIEVLLERELAELSAELDKDLRAIETRLPSPKSSPAPRRAPEQRPPAGPASAWSSSYPHAPYLGSARSLSPHKMADGGSPFLGRRDFVYPSSTRDPSASNGGGSPARREEKKRKAARLKFDFQAQSPKELTLQKGDIVYIHKEVDKNWLEGEHHGRLGIFPANYVEVLPADEIPKPIKPPTYQVLEYGEAVAQYTFKGDLEVELSFRKGEHICLIRKVNENWYEGRITGTGRQGIFPASYVQVSREPRLRLCDDGPQLPTSPRLTAAARSARHPSSPSALRSPADPIDLGGQTSPRRTGFSFPTQEPRPQTQNLGTPGPALSHSRGPSHPLDLGTSSPNTSQIHWTPYRAMYQYRPQNEDELELREGDRVDVMQQCDDGWFVGVSRRTQKFGTFPGNYVAPV 2D8A8718E0367BE9 safb2_human Scaffold attachment factor B2 SAFB2 KIAA0138 protein protein human Homo sapiens Human MAETLPGSGDSGPGTASLGPGVAETGTRRLSELRVIDLRAELKKRNLDTGGNKSVLMERLKKAVKEEGQDPDEIGIELEATSKKSAKRCVKGLKMEEEGTEDNGLEDDSRDGQEDMEASLENLQNMGMMDMSVLDETEVANSSAPDFGEDGTDGLLDSFCDSKEYVAAQLRQLPAQPPEHAVDGEGFKNTLETSSLNFKVTPDIEESLLEPENEKILDILGETCKSEPVKEESSELEQPFAQDTSSVGPDRKLAEEEDLFDSAHPEEGDLDLASESTAHAQSSKADSLLAVVKREPAEQPGDGERTDCEPVGLEPAVEQSSAASELAEASSEELAEAPTEAPSPEARDSKEDGRKFDFDACNEVPPAPKESSTSEGADQKMSSFKEEKDIKPIIKDEKGRVGSGSGRNLWVSGLSSTTRATDLKNLFSKYGKVVGAKVVTNARSPGARCYGFVTMSTSDEATKCISHLHRTELHGRMISVEKAKNEPAGKKLSDRKECEVKKEKLSSVDRHHSVEIKIEKTVIKKEEKIEKKEEKKPEDIKKEEKDQDELKPGPTNRSRVTKSGSRGMERTVVMDKSKGEPVISVKTTSRSKERSSKSQDRKSESKEKRDILSFDKIKEQRERERQRQREREIRETERRREREQREREQRLEAFHERKEKARLQRERLQLECQRQRLERERMERERLERERMRVERERRKEQERIHREREELRRQQEQLRYEQERRPGRRPYDLDRRDDAYWPEGKRVAMEDRYRADFPRPDHRFHDFDHRDRGQYQDHAIDRREGSRPMMGDHRDGQHYGDDRHGHGGPPERHGRDSRDGWGGYGSDKRLSEGRGLPPPPRGGRDWGEHNQRLEEHQARAWQGAMDAGAASREHARWQGGERGLSGPSGPGHMASRGGVAGRGGFAQGGHSQGHVVPGGGLEGGGVASQDRGSRVPHPHPHPPPYPHFTRRY 084343934F8B3196 safb2-esr1-1 3 7 unspecified role unspecified role bait bait dbd_hinge 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) 11 unspecified role unspecified role prey prey in vitro In vitro direct interaction direct interaction As shown in Fig. 3A, SAFB2 can bind to ER in GST pull-down assays, suggesting a direct interaction. Fig. 3A safb2-esr1-2 5 11 unspecified role unspecified role bait bait in vitro In vitro 7 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) physical association physical association Subsequently we used this antibody to immunoprecipitate SAFB2 from MCF-7 lysates, and immunoblotting with anti-ER antibodies confirmed the in vivo interaction between SAFB2 and ER (Fig. 3C). Fig. 3C safb1-safb2-1 2 11 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 24 22 physical association physical association SAFB1 and SAFB2 interact in yeast two-hybrid assays Fig. 4A Amino acids 719-808 are the major interaction region and amino acids 599-720 are a region of minor interaction. safb1-safb2-2 4 8 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role prey prey 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 physical association physical association To confirm the interaction between SAFB1 and SAFB2 in vivo, we coimmunoprecipitated differentially tagged proteins from CV-1 cells. Fig. 4B Amino acids 719-808 are the major interaction region and amino acids 519-720 are a region of minor interaction. safb1-safb2-3 6 11 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association SAFB1 and SAFB2 proteins interact in MCF-7 RTA-HA-SAFB1 cells. Fig. 4C safb2-sorbs3-1 5 9 unspecified role unspecified role prey prey 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 11 unspecified role unspecified role bait bait in vitro In vitro physical association physical association First we transfected MCF-7 cells with GFP-vinexin {beta}, precipitated SAFB2 from the lysates, and subsequently immunoblotted the membrane with anti-vinexin antibodies. Fig. 5B safb2-sorbs3-2 1 9 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 11 unspecified role unspecified role neutral component neutral component in vitro In vitro colocalization colocalization Overlay of GFP-vinexin {beta} and SAFB2 staining revealed colocalization in discrete focal points in a subset of cells. Fig. 5C safb2-sorbs3-3 5 10 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 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 11 unspecified role unspecified role bait bait in vitro In vitro association association Finally, we immunoprecipitated SAFB2 from nontransfected MCF-7 cells and immunoblotted the membrane with anti-vinexin antibodies. As shown in Fig. 5D, we detected not only an interaction with vinexin {beta} but also with vinexin {alpha}. Fig. 5D