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/ metz-2006-2 The yeast Arr4p ATPase binds the chloride transporter Gef1p when copper is available in the cytosol. 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 Metz J., Wachter A., Schmidt B., Bujnicki JM., Schwappach B. BioCreative - Critical Assessment of Information Extraction systems in Biology In some studies BCS (bathophenantroline) at 3mM was included in the yeast two hybrid selection media. b.schwappach@zmbh.uni-heidelberg.de Yeast two hybrid genomic library J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation metz-2006-3 The yeast Arr4p ATPase binds the chloride transporter Gef1p when copper is available in the cytosol. in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Metz J., Wachter A., Schmidt B., Bujnicki JM., Schwappach B. BioCreative - Critical Assessment of Information Extraction systems in Biology b.schwappach@zmbh.uni-heidelberg.de Yeast two hybrid genomic library J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation metz-2006-1 The yeast Arr4p ATPase binds the chloride transporter Gef1p when copper is available in the cytosol. 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 Metz J., Wachter A., Schmidt B., Bujnicki JM., Schwappach B. BioCreative - Critical Assessment of Information Extraction systems in Biology b.schwappach@zmbh.uni-heidelberg.de Yeast two hybrid genomic library J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation metz-2006-4 The yeast Arr4p ATPase binds the chloride transporter Gef1p when copper is available in the cytosol. in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Metz J., Wachter A., Schmidt B., Bujnicki JM., Schwappach B. BioCreative - Critical Assessment of Information Extraction systems in Biology b.schwappach@zmbh.uni-heidelberg.de Yeast two hybrid genomic library J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation metz-2006-5 The yeast Arr4p ATPase binds the chloride transporter Gef1p when copper is available in the cytosol. in vitro In vitro pull down pull down predetermined predetermined participant predetermined Metz J., Wachter A., Schmidt B., Bujnicki JM., Schwappach B. BioCreative - Critical Assessment of Information Extraction systems in Biology Proteins were identified by Coomassie blue staining after SDS PAGE . Detection of protiens by SDS PAGE b.schwappach@zmbh.uni-heidelberg.de Yeast two hybrid genomic library J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation metz-2006-6 The yeast Arr4p ATPase binds the chloride transporter Gef1p when copper is available in the cytosol. in vitro In vitro bn-page blue native page bn-page predetermined predetermined participant predetermined Metz J., Wachter A., Schmidt B., Bujnicki JM., Schwappach B. BioCreative - Critical Assessment of Information Extraction systems in Biology b.schwappach@zmbh.uni-heidelberg.de Yeast two hybrid genomic library J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation gef1_yeast Anion/proton exchange transporter GEF1 CLCY1 GEF1 YJR040W J1616 Voltage-gated chloride channel ClC-Y1 ClC-A CLC protein GEF1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MPTTYVPINQPIGDGEDVIDTNRFTNIPETQNFDQFVTIDKIAEENRPLSVDSDREFLNSKYRHYREVIWDRAKTFITLSSTAIVIGCIAGFLQVFTETLVNWKTGHCQRNWLLNKSFCCNGVVNEVTSTSNLLLKRQEFECEAQGLWIAWKGHVSPFIIFMLLSVLFALISTLLVKYVAPMATGSGISEIKVWVSGFEYNKEFLGFLTLVIKSVALPLAISSGLSVGKEGPSVHYATCCGYLLTKWLLRDTLTYSSQYEYITAASGAGVAVAFGAPIGGVLFGLEEIASANRFNSSTLWKSYYVALVAITTLKYIDPFRNGRVILFNVTYDRDWKVQEIPIFIALGIFGGLYGKYISKWNINFIHFRKMYLSSWPVQEVLFLATLTALISYFNEFLKLDMTESMGILFHECVKNDNTSTFSHRLCQLDENTHAFEFLKIFTSLCFATVIRALLVVVSYGARVPAGIFVPSMAVGATFGRAVSLLVERFISGPSVITPGAYAFLGAAATLSGITNLTLTVVVIMFELTGAFMYIIPLMIVVAITRIILSTSGISGGIADQMIMVNGFPYLEDEQDEEEEETLEKYTAEQLMSSKLITINETIYLSELESLLYDSASEYSVHGFPITKDEDKFEKEKRCIGYVLKRHLASKIMMQSVNSTKAQTTLVYFNKSNEELGHRENCIGFKDIMNESPISVKKAVPVTLLFRMFKELGCKTIIVEESGILKGLVTAKDILRFKRIKYREVHGAKFTYNEALDRRCWSVIHFIIKRFTTNRNGNVI 544C751402D32E19 get3_yeast ATPase GET3 YDL100C GET3 ARR4 D2371 Arsenical pump-driving ATPase Arsenite-stimulated ATPase Guided entry of tail-anchored proteins 3 Golgi to ER traffic protein 3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MDLTVEPNLHSLITSTTHKWIFVGGKGGVGKTTSSCSIAIQMALSQPNKQFLLISTDPAHNLSDAFGEKFGKDARKVTGMNNLSCMEIDPSAALKDMNDMAVSRANNNGSDGQGDDLGSLLQGGALADLTGSIPGIDEALSFMEVMKHIKRQEQGEGETFDTVIFDTAPTGHTLRFLQLPNTLSKLLEKFGEITNKLGPMLNSFMGAGNVDISGKLNELKANVETIRQQFTDPDLTTFVCVCISEFLSLYETERLIQELISYDMDVNSIIVNQLLFAENDQEHNCKRCQARWKMQKKYLDQIDELYEDFHVVKMPLCAGEIRGLNNLTKFSQFLNKEYNPITDGKVIYELEDKE F795C359B5A4A461 aakg_yeast 5'-AMP-activated protein kinase subunit gamma SNF4 CAT3 Regulatory protein CAT3 SCI1 Sucrose non-fermenting protein 4 YGL115W protein protein yeast Saccharomyces cerevisiae Baker's yeast MKPTQDSQEKVSIEQQLAVESIRKFLNSKTSYDVLPVSYRLIVLDTSLLVKKSLNVLLQNSIVSAPLWDSKTSRFAGLLTTTDFINVIQYYFSNPDKFELVDKLQLDGLKDIERALGVDQLDTASIHPSRPLFEACLKMLESRSGRIPLIDQDEETHREIVVSVLTQYRILKFVALNCRETHFLKIPIGDLNIITQDNMKSCQMTTPVIDVIQMLTQGRVSSVPIIDENGYLINVYEAYDVLGLIKGGIYNDLSLSVGEALMRRSDDFEGVYTCTKNDKLSTIMDNIRKARVHRFFVVDDVGRLVGVLTLSDILKYILLGSN 51B387E346EE9561 snf1_yeast Carbon catabolite-derepressing protein kinase SNF1 CAT1 CCR1 GLC2 PAS14 Sucrose nonfermentating protein 1 D8035.20 YDR477W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSSNNNTNTAPANANSSHHHHHHHHHHHHHGHGGSNSTLNNPKSSLADGAHIGNYQIVKTLGEGSFGKVKLAYHTTTGQKVALKIINKKVLAKSDMQGRIEREISYLRLLRHPHIIKLYDVIKSKDEIIMVIEYAGNELFDYIVQRDKMSEQEARRFFQQIISAVEYCHRHKIVHRDLKPENLLLDEHLNVKIADFGLSNIMTDGNFLKTSCGSPNYAAPEVISGKLYAGPEVDVWSCGVILYVMLCRRLPFDDESIPVLFKNISNGVYTLPKFLSPGAAGLIKRMLIVNPLNRISIHEIMQDDWFKVDLPEYLLPPDLKPHPEEENENNDSKKDGSSPDNDEIDDNLVNILSSTMGYEKDEIYESLESSEDTPAFNEIRDAYMLIKENKSLIKDMKANKSVSDELDTFLSQSPPTFQQQSKSHQKSQVDHETAKQHARRMASAITQQRTYHQSPFMDQYKEEDSTVSILPTSLPQIHRANMLAQGSPAASKISPLVTKKSKTRWHFGIRSRSYPLDVMGEIYIALKNLGAEWAKPSEEDLWTIKLRWKYDIGNKTNTNEKIPDLMKMVIQLFQIETNNYLVDFKFDGWESSYGDDTTVSNISEDEMSTFSAYPFLHLTTKLIMELAVNSQSN F5C63565C986C4E3 gef1-get3-1 Directed yeast 2 hybrid analysis reveals interaction between GEF1protein and GET3 protein 494111 N/A 494117 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 494118 unspecified role unspecified role prey prey physical association physical association false false false Figure 1A Figure 4A Interaction was not seen when 3mM BCS (bathophenantroline sulphate) (a copper chelating agent) was present in the yeast culture media. snf1-snf4 Directed Yeast 2 hybrid analysis reveals interaction between the SNF1 protein and SNF4 protein 494111 N/A 494119 unspecified role unspecified role prey prey N/A 494120 unspecified role unspecified role bait bait physical association physical association false false false BCS was not used in the selection media in this study. Figure 1 A Positive control used in directed yeast two hybrid assay. get3-get3 Directed yeast 2 hybrid analysis reveals homotypic interaction of the GET3 protein 494111 N/A 494118 unspecified role unspecified role bait bait Q12154:p.[Cys285Thr;Cys288Thr] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false N/A 494118 unspecified role unspecified role prey prey physical association physical association false false false BCS was not used in the selcetion media in this study. Figure 6B gef1-get3-2 Coimmunoprecipitation analysis reveals interaction between GEF1 protein and GET3 protein 494112 N/A 494118 unspecified role unspecified role bait bait yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast N/A 494117 unspecified role unspecified role prey prey n terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association false false false Figure 1B gef1-get3-5 Coimmunoprecipitation analysis reveals interaction between GEF1 protein and overexpressed GET3 protein 494112 N/A 494118 unspecified role unspecified role bait bait yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast N/A 494117 unspecified role unspecified role prey prey n terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association false false false Figure 1B Over-expression of Arr4p Q12154 gef1-get3 Yeast 2 hybrid analysis reveals interaction between GEF1protein and GET3 protein 494113 N/A 494118 unspecified role unspecified role prey prey N/A 494117 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false gef1-get3-3 Coimmunoprecipitation analysis reveals interaction between GEF1 protein and GET3 protein 494114 N/A 494118 unspecified role unspecified role prey prey region Protein C tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast N/A 494117 unspecified role unspecified role bait bait n terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association false false false Figure 1C Gef1 protein was overexpressed in the yeast strain analysed in this study. gef1-get3-4 Invitro binding analysis reveals interaction between GEF1protein and GET3 protein 494115 N/A 494117 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region IGg A protein tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 494118 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) physical association physical association false false false Figure 2 B and 3A This interaction was only observed when yeast lysate, or trace mineral solution or CuSo4 (160nm) and ascorbate (10 micro M) was present in the binding assay reaction buffer. get3-get3-2 Blue native gel electrophoresis analysis reveals homotypic interaction of the GET3 protein 494116 N/A 494118 unspecified role unspecified role neutral component neutral component N/A 494118 unspecified role unspecified role neutral component neutral component Q12154:p.[Cys285Thr;Cys288Thr] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 6C