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 PDZ protein Canoe/AF-6 links Ras-MAPK, Notch and Wingless/Wnt signaling pathways by directly interacting with Ras, Notch and Dishevelled. The PDZ protein Canoe/AF-6 links Ras-MAPK, Notch and Wingless/Wnt signaling pathways by directly interacting with Ras, Notch and Dishevelled. PLoS ONE 2006 imex curation Carmena A., Speicher S., Baylies M. Regions of the proteins based on the information and reference provided in the publication (PMID: 8596950). acarmena@umh.es 01-Feb-2008: Contacted by JYOTI. 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 Carmena A., Speicher S., Baylies M. Regions of the proteins based on the information and reference provided in the publication (PMID: 8596950). acarmena@umh.es Accepted 2008-FEB-9 by DAVET. 01-Feb-2008: Contacted by JYOTI. PLoS ONE 2006 davet Only protein-protein interactions imex curation The PDZ protein Canoe/AF-6 links Ras-MAPK, Notch and Wingless/Wnt signaling pathways by directly interacting with Ras, Notch and Dishevelled. The PDZ protein Canoe/AF-6 links Ras-MAPK, Notch and Wingless/Wnt signaling pathways by directly interacting with Ras, Notch and Dishevelled. PLoS ONE 2006 imex curation Carmena A., Speicher S., Baylies M. Regions of the proteins based on the information and reference provided in the publication (PMID: 8596950). acarmena@umh.es 01-Feb-2008: Contacted by JYOTI. davet Only protein-protein interactions in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Carmena A., Speicher S., Baylies M. Regions of the proteins based on the information and reference provided in the publication (PMID: 8596950). acarmena@umh.es Accepted 2008-FEB-9 by DAVET. 01-Feb-2008: Contacted by JYOTI. PLoS ONE 2006 davet Only protein-protein interactions imex curation The PDZ protein Canoe/AF-6 links Ras-MAPK, Notch and Wingless/Wnt signaling pathways by directly interacting with Ras, Notch and Dishevelled. The PDZ protein Canoe/AF-6 links Ras-MAPK, Notch and Wingless/Wnt signaling pathways by directly interacting with Ras, Notch and Dishevelled. PLoS ONE 2006 imex curation Carmena A., Speicher S., Baylies M. Regions of the proteins based on the information and reference provided in the publication (PMID: 8596950). acarmena@umh.es 01-Feb-2008: Contacted by JYOTI. davet Only protein-protein interactions drome-embryo drosophilla melanogaster embryo embryo Embryo Defined as the development stage prior to the initiation of tissue differentiation. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Carmena A., Speicher S., Baylies M. Proteins extracts from 0 to 7 h old fly embryos were used. Regions of the proteins based on the information and reference provided in the publication (PMID: 8596950). acarmena@umh.es Accepted 2008-FEB-9 by DAVET. 01-Feb-2008: Contacted by JYOTI. PLoS ONE 2006 davet Only protein-protein interactions imex curation dsh_drome Segment polarity protein dishevelled dsh CG18361 protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MDADRGGGQETKVIYHIDDETTPYLVKIPIPSAQVTLRDFKLVLNKQNNNYKYFFKSMDADFGVVKEEIADDSTILPCFNGRVVSWLVSADGTNQSDNCSELPTSECELGMGLTNRKLQQQQQQHQQQQQQQQQQHQQQQQQQQQQVQPVQLAQQQQQQVLHHQKMMGNPLLQPPPLTYQSASVLSSDLDSTSLFGTESELTLDRDMTDYSSVQRLQVRKKPQRRKKRAPSMSRTSSYSSITDSTMSLNIITVSINMEAVNFLGISIVGQSNRGGDGGIYVGSIMKGGAVALDGRIEPGDMILQVNDVNFENMTNDEAVRVLREVVQKPGPIKLVVAKCWDPNPKGYFTIPRTEPVRPIDPGAWVAHTQALTSHDSIIADIAEPIKERLDQNNLEEIVKAMTKPDSGLEIRDRMWLKITIPNAFIGADAVNWVLENVEDVQDRREARRIVSAMLRSNYIKHTVNKLTFSEQCYYVVNEERNPNLLGRGHLHPHQLPHGHGGHALSHADTESITSDIGPLPNPPIYMPYSATYNPSHGYQPIQYGIAERHISSGSSSSDVLTSKDISASQSDITSVIHQANQLTIAAHGSNKSSGSSNRGGGGGGGGGGNNTNDQDVSVFNYVL 0BA24ED350666CF5 notch_drome Neurogenic locus Notch protein N CG3936 protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MQSQRSRRRSRAPNTWICFWINKMHAVASLPASLPLLLLTLAFANLPNTVRGTDTALVAASCTSVGCQNGGTCVTQLNGKTYCACDSHYVGDYCEHRNPCNSMRCQNGGTCQVTFRNGRPGISCKCPLGFDESLCEIAVPNACDHVTCLNGGTCQLKTLEEYTCACANGYTGERCETKNLCASSPCRNGATCTALAGSSSFTCSCPPGFTGDTCSYDIEECQSNPCKYGGTCVNTHGSYQCMCPTGYTGKDCDTKYKPCSPSPCQNGGICRSNGLSYECKCPKGFEGKNCEQNYDDCLGHLCQNGGTCIDGISDYTCRCPPNFTGRFCQDDVDECAQRDHPVCQNGATCTNTHGSYSCICVNGWAGLDCSNNTDDCKQAACFYGATCIDGVGSFYCQCTKGKTGLLCHLDDACTSNPCHADAICDTSPINGSYACSCATGYKGVDCSEDIDECDQGSPCEHNGICVNTPGSYRCNCSQGFTGPRCETNINECESHPCQNEGSCLDDPGTFRCVCMPGFTGTQCEIDIDECQSNPCLNDGTCHDKINGFKCSCALGFTGARCQINIDDCQSQPCRNRGICHDSIAGYSCECPPGYTGTSCEININDCDSNPCHRGKCIDDVNSFKCLCDPGYTGYICQKQINECESNPCQFDGHCQDRVGSYYCQCQAGTSGKNCEVNVNECHSNPCNNGATCIDGINSYKCQCVPGFTGQHCEKNVDECISSPCANNGVCIDQVNGYKCECPRGFYDAHCLSDVDECASNPCVNEGRCEDGINEFICHCPPGYTGKRCELDIDECSSNPCQHGGTCYDKLNAFSCQCMPGYTGQKCETNIDDCVTNPCGNGGTCIDKVNGYKCVCKVPFTGRDCESKMDPCASNRCKNEAKCTPSSNFLDFSCTCKLGYTGRYCDEDIDECSLSSPCRNGASCLNVPGSYRCLCTKGYEGRDCAINTDDCASFPCQNGGTCLDGIGDYSCLCVDGFDGKHCETDINECLSQPCQNGATCSQYVNSYTCTCPLGFSGINCQTNDEDCTESSCLNGGSCIDGINGYNCSCLAGYSGANCQYKLNKCDSNPCLNGATCHEQNNEYTCHCPSGFTGKQCSEYVDWCGQSPCENGATCSQMKHQFSCKCSAGWTGKLCDVQTISCQDAADRKGLSLRQLCNNGTCKDYGNSHVCYCSQGYAGSYCQKEIDECQSQPCQNGGTCRDLIGAYECQCRQGFQGQNCELNIDDCAPNPCQNGGTCHDRVMNFSCSCPPGTMGIICEINKDDCKPGACHNNGSCIDRVGGFECVCQPGFVGARCEGDINECLSNPCSNAGTLDCVQLVNNYHCNCRPGHMGRHCEHKVDFCAQSPCQNGGNCNIRQSGHHCICNNGFYGKNCELSGQDCDSNPCRVGNCVVADEGFGYRCECPRGTLGEHCEIDTLDECSPNPCAQGAACEDLLGDYECLCPSKWKGKRCDIYDANYPGWNGGSGSGNDRYAADLEQQRAMCDKRGCTEKQGNGICDSDCNTYACNFDGNDCSLGINPWANCTANECWNKFKNGKCNEECNNAACHYDGHDCERKLKSCDSLFDAYCQKHYGDGFCDYGCNNAECSWDGLDCENKTQSPVLAEGAMSVVMLMNVEAFREIQAQFLRNMSHMLRTTVRLKKDALGHDIIINWKDNVRVPEIEDTDFARKNKILYTQQVHQTGIQIYLEIDNRKCTECFTHAVEAAEFLAATAAKHQLRNDFQIHSVRGIKNPGDEDNGEPPANVKYVITGIILVIIALAFFGMVLSTQRKRAHGVTWFPEGFRAPAAVMSRRRRDPHGQEMRNLNKQVAMQSQGVGQPGAHWSDDESDMPLPKRQRSDPVSGVGLGNNGGYASDHTMVSEYEEADQRVWSQAHLDVVDVRAIMTPPAHQDGGKHDVDARGPCGLTPLMIAAVRGGGLDTGEDIENNEDSTAQVISDLLAQGAELNATMDKTGETSLHLAARFARADAAKRLLDAGADANCQDNTGRTPLHAAVAADAMGVFQILLRNRATNLNARMHDGTTPLILAARLAIEGMVEDLITADADINAADNSGKTALHWAAAVNNTEAVNILLMHHANRDAQDDKDETPLFLAAREGSYEACKALLDNFANREITDHMDRLPRDVASERLHHDIVRLLDEHVPRSPQMLSMTPQAMIGSPPPGQQQPQLITQPTVISAGNGGNNGNGNASGKQSNQTAKQKAAKKAKLIEGSPDNGLDATGSLRRKASSKKTSAASKKAANLNGLNPGQLTGGVSGVPGVPPTNSAAQAAAAAAAAVAAMSHELEGSPVGVGMGGNLPSPYDTSSMYSNAMAAPLANGNPNTGAKQPPSYEDCIKNAQSMQSLQGNGLDMIKLDNYAYSMGSPFQQELLNGQGLGMNGNGQRNGVGPGVLPGGLCGMGGLSGAGNGNSHEQGLSPPYSNQSPPHSVQSSLALSPHAYLGSPSPAKSRPSLPTSPTHIQAMRHATQQKQFGGSNLNSLLGGANGGGVVGGGGGGGGGVGQGPQNSPVSLGIISPTGSDMGIMLAPPQSSKNSAIMQTISPQQQQQQQQQQQQQHQQQQQQQQQQQQQQQQQLGGLEFGSAGLDLNGFCGSPDSFHSGQMNPPSIQSSMSGSSPSTNMLSPSSQHNQQAFYQYLTPSSQHSGGHTPQHLVQTLDSYPTPSPESPGHWSSSSPRSNSDWSEGVQSPAANNLYISGGHQANKGSEAIYI DIP protein P07207 original sequence version: 144 DIP protein P07207 original sequence version: 184 0EAE23F426FECD7B q24279_drome cno CG42312 protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MSHDKKMLDREAVRSVIQQWNANRLDLFALSEPDENLLFHGVMRFYFQDAGQKVATKCIRVASDATVTDVIDTLIEKFRPDMRMLSVPNYALYEVHANGEERRLNADEKPLLVQLNWHIDDREGRFLLKNIDQKTTPIEQTDLNFKRKLSKREKKEQKKKDKMAKLSSDPPTSNGSHLGLGNGVGASSGSAHMNGNAGGTGGDSSDAVAGKLYTELPETSFTRSISNPEAVMRRRRQQKLEKKLQQFRSRDGGPDTGGTLKIYGESLCQDVPYKTLLLSIRDCAQAVVREMLTKYGLEKADPLHYCLVQVNSDGTEYILDDDECPLSILMNHPTSRGSIMFHVRRRPADSQPRRRKKKPLGAANGANHISGDREGPVLVEVTHSGDGGRRIKLGSDPVEVGSANTNCLQLFGPSIQPRHCLISLLEGVCTVTPLHTDALTFVNGHHISQPTILHNGSVVMFGRVASYRFLDSPTDGRYNLALSQSQLDSACLYERYRASTHYRPELQPTERAHKLTMFLHHVANLVYSVVQEQYTDPRILAFWMANSSEFLHFLKSDRHISAFSVQAQEVLAEAVQTAFRNLVNCFRLELSQTLNQFLSENIDHDSAAGLVLTVLGSAMALLRRCRVNAALTIQLFSQLFHYINVICFNTIVANSQMCTGDWGKVMTERLQLLELWAERQGLELAADCHLAKINQCAQFLQAPKSSVEEIQQLACSCFRLNSLQMAALLQQEKLPRNLVDTAIRMAESVADELTRADGREVRLEESPELHLALLLPDDGFSCDVVRGIPTGLVDFLNPLQQQGMCRLAAQPTSIGLWTVYMHQFNARSSSAMSNKLPQPELQLIKLHKNSNGMGLSIVASKGAGQEKLGIYIKSVVPGGAADADGRLQAGDQLLRVDGQSLIGITQERAADYLVRTGPVVSLEVAKQGAIYHGLATLLQQPSPVIQRGNRRMSERDLTRMGGAESTKSLGPLIANNHPNHSLSLSQHLNNSHNNTLGNSNNSSIINPLAAHLPNSKSVPALHHHTGSGTISLANSKSRSTHSLHNNTSGMGGIGGAAAGGMLGQPNGSQNNANGNGNGNEQGFYQNLSVYRAQNQSQPILNERPPIAAHAAMNAYNGSSPLAPQQQPQQQQQSPYQQQQQQQHMQANANLPPTRPVSAYYHSQQSAQQQLQQQQQQQQQHSLQQQQFALSSGNLNGQQQQQQQHQHQLTLNNRTKSQQNFQHTLRMQQMMAPSMPNISNMYHHQQQHQQQLPLQQQQQQQQQPLMSSSQSMQNVNDFAGGYQNGSLEYRRSQLHDPSTLYEIQQQQLQQQQQQQQQQQQQQQASPNFIALPPKPLGSLQSPNKPNVPPSTAPKPQQQQQRYLGQSLPAEDKPPLPPTATHPLFKATQQIAPGMNYVASTLDPPKGSYVASNQGNNRPLHSGSNPWEREEREKDLEMRREHIRQWREQQISELSQIVSRSPMQEEQLKTLILERDFERRAQELQEQEEQDQEQQYDKENVQELFRLAGGGQVSAIQTPITSYRQTEIKLAEMPDSNSLVDSVPPQPPAPTAQPLSSNTQQPKSILKHNRYSEGGVGPSGAPSSPSKSQKSASFANERHLHTEHPISNLAKELNQLTMLDKDNNNETLDAVVPPPPPPERNSSYLIMSQQKLRGSTGNATSSMGLLKTATSNQAAAAVEIKKASLLNTQTNNNNNLSGSLNNNTMHGSPLSAMELNAAYVSATGTGIIGGLGTPPPPPPLMQRDNKRVSFHDEENNFVSGNSQQQQLQQQYIMDNYGMEHQDLDTIREDTSYLQHNLDASMLPSMPATPDAALWNTSVQSTPGVIGAQEVYRDPRTRRLAEKQQQQQQQRAGDAVPEKLSFKEKMKMFALESGEDKTPKDKLKISRAQRDIDAVH E958E3470783CDDA cno-dsh 1 4 unspecified role unspecified role bait bait dix and pdz domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain dep domain sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1573993]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 638-638, sequence length 623 [EBI-1573993]286-638 6 unspecified role unspecified role prey prey ras associated domai sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain pdz sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 12 9 13 10 physical association physical association 5D The Cno-Dsh interaction, however, mapped to two different domains of Cno: the PDZ domain and the Ras-binding domain (Fig. 5C and 5D). To distinguish which domains in Dsh bound the Ras-associating and PDZ motifs of Cno, different Dsh constructs were tested. A Dsh fragment containing the DIX (DIshevelled, aXin) and PDZ domains (DSH-DIZ) only bound Cno constructs containing the Ras-binding domain, as did the Ras positive control (Fig. 5C and 5D). In contrast, the Dsh fragment containing the DEP (Dsh, egl-10, pleckstrin) domain (DSH-DEP) only bound Cno constructs containing the PDZ domain plus some of the adjacent Myosin-V (Myo)-like domain n-cno 3 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait physical association physical association 5A Cno coimmunoprecipitates with both N and Dsh The co-IPs from embryonic lysates showed different sizes of the Cno proteins coimmunoprecipitated with either Dsh or N dsh-cno 3 4 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association 5A Cno coimmunoprecipitates with both N and Dsh The co-IPs from embryonic lysates showed different sizes of the Cno proteins coimmunoprecipitated with either Dsh or N n-cno-1 2 5 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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 5B Cno and N proteins directly interacted in vitro cno-dsh-1 2 6 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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 7I We found that addition of increasing amounts of Dsh led to reduced amounts of RasV12 (RasAct) co-immunoprecipitated with Cno The addition of increasing amounts of Dsh leads to a decrease in the amount of RasV12 co-IP with Cno. The addition of increasing amounts of Dsh leads to a decrease in the amount of RasV12 co-IP with Cno. An unique entry could however not be assigned to Ras protein. cno-n 1 5 unspecified role unspecified role prey prey icn2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 6 unspecified role unspecified role bait bait myo like and pdz sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain myo like and pdz sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 34 37 physical association physical association 5C and 5D To characterize the domains implicated in the binding, diverse deletion constructs of Cno, N and Dsh were tested in yeast two-hybrid assays (Fig. 5C and 5D). Only Cno constructs comprising the PDZ domain strongly interacted with both proximal (Notch-ICN1) and distal (Notch-ICN2) regions of the N intracellular domain