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/ martin-2005-2 Tea4p links microtubule plus ends with the formin for3p in the establishment of cell polarity. schpm Schizosaccharomyces pombe (fission yeast) Fission yeast anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Martin SG., McDonald WH., Yates JR., Chang F. fc99@columbia.edu Dev. Cell (1534-5807) 2005 Only protein-protein interactions imex curation martin-2005-1 Tea4p links microtubule plus ends with the formin for3p in the establishment of cell polarity. 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 Martin SG., McDonald WH., Yates JR., Chang F. fc99@columbia.edu Dev. Cell (1534-5807) 2005 Only protein-protein interactions imex curation martin-2005-3 Tea4p links microtubule plus ends with the formin for3p in the establishment of cell polarity. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Martin SG., McDonald WH., Yates JR., Chang F. fc99@columbia.edu Dev. Cell (1534-5807) 2005 Only protein-protein interactions imex curation martin-2005-4 Tea4p links microtubule plus ends with the formin for3p in the establishment of cell polarity. schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 2h fragment pooling two hybrid fragment pooling approach 2h fragment pooling predetermined predetermined participant predetermined Martin SG., McDonald WH., Yates JR., Chang F. fc99@columbia.edu Dev. Cell (1534-5807) 2005 Only protein-protein interactions imex curation tea4_schpo Tip elongation aberrant protein Tea4 SPBC1706.01 tea4 wsh3 Cell polarity protein tea4 Win1-interacting SH3 domain protein protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MLHMNSASSADSMEIMESHFDPTQQNDSTIIESRYSPEEYLEQSFEIQRIISGENSEPQTVASQEISDSQEEDTTLTSSQFEDCGTEYNEVVEDDEFRSEDEDDFMDEEEEYALYEAELSSSPSIHEEVIDCNFVHAIRGFEATVEGQVDATKGDMMILLDDSNSYWWLVKMCKNLAIGYLPAEYIETPSERLARLNKYKNSETSNSQQSVTLPPLDIVEKTLEAPSPNFRIKRVTFTCSSNSSDDEMDSENDYEAMVNRTVAENGLEIEFSDSSDSSLSAEYRSESEDHVTDSPAYVDLTELEGGFNQFNSTSFQSTSPLGLEIVETEINGSSTTADSKNSHSPYSKFSSAYPDAENSNISKINISIAGNKELYGNATQSDPSLYSTWIANKHKTASSATVDSPLRRSLSVDAMQSNASFSSYSSTSNTDKSLRPSSYSAVSESSNFTHDVSRDNKEISLNAPKSIIVSQSDSFDTSNVTQDAPNDVEKEPISGQMPNNLSVQSLKQLEVYPIRHSVSIEMPSEKLLSPRLYSSSTPSSPTKGFQKDDEEDSENRKQADKVELSPSSLLRQMSLPVDSSSQSDAQCTTSSVYITAERKAFSQSSIDLSTLSNHHVNNEINRRSFAGGFTSLADELSEMRELLHESPAPLECNEEMVIPTPELDASSAIPSSSISHDEDLLPRKNTEESTSSSSFSSLITSPASLQYDENPFKQSVVAELNNNSSSVPFVDSAHASDIHAYDNDHVSTKNKEFNRRLREFILDPDSLSGLYWSVKSAGVRASRRVSRNIEGESVSSDLDDIFANVLKGLSDEMASLLNTNR tea1_schpo Tip elongation aberrant protein 1 Cell polarity protein tea1 Altered polarity protein 8 tea1 SPCC1223.06 alp8 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSFLFKRNKGSAHKPTKPNFSKTSTTPSTSQLKHSHESNVKMSTSTVTEHRKKPTGSGSHITASPWSKLTVRGSSNVLPRYSHASHLYAEGGQEIYIFGGVASDSQPKNDLWVLNLATSQFTSLRSLGETPSPRLGHASILIGNAFIVFGGLTNHDVADRQDNSLYLLNTSSLVWQKANASGARPSGRYGHTISCLGSKICLFGGRLLDYYFNDLVCFDLNNLNTSDSRWELASVVNDPPPARAGHVAFTFSDKLYIFGGTDGANFFNDLWCYHPKQSAWSKVETFGVAPNPRAGHAASVVEGILYVFGGRASDGTFLNDLYAFRLSSKHWYKLSDLPFTPSPRSSHTLSCSGLTLVLIGGKQGKGASDSNVYMLDTSRFRLGSVPTTSGRQRNTSFFSNSTGNTNPSAFNGLLTSSRIPSYNGSKVRSTSHPSRQQYIGSSNSRFNTRHQTISTPVSGRASNDLPSPVVPTRSNSSSILQPSYNLNSHSSDRRNTNDDDQSSLNSQQLSNQAKAQGEVSPTLSFVPSSHSMEQGNGSVASANNAQSEAATRSINSISEVSEVRFPEQSSVKTVDERKSLDGRITSVTLETLVEKYSELSKQQIVEWFKSKLYEILRDSASKIDSLTEKLKVANAEKNAALCEAALEKVPLAKHNKLSDGTFSTPDKENVQSTNDAHIMQENFSLHKALEVMRETSSDLDKQLKDATASQKELIVQTSSFQKELVEERERHNAISKRLQEIESLYRDRELLVTNLEDQLVDQTVTINKFAFERDQFRERSMGFENTIKDLTRKMEATDMLNVSLHESLRSVQTENSELVTEMALLKAELVKKQAIIDANANIYDKLTADHTNYETVSADINQNLKETLDKLLNGSSDFKNNEIELLHDQIRITNAKLEKREKLINASKYIEDTLRSEIQEAAEKVSNLEFSNFNLKEENSNMQLQLMKALEQRNTGAKQLVNLRMQLSTATSELDMLKLKLRTTALALEESPDDYSDILSILRADMSPFHDLHKQCGVLIDTLNGVKRGFGIFEKKFTDYHKFLENISDKLKSEEDTSLETPIHENQSIQSDQIKEVGEVLSAIKSLSDSVMLLKNQIDDLAKEKLPLSSSDDEKVNIKEKTDFMKLLVKSGLSNPPAKEPVHDNEN tip1_schpo Tip elongation protein 1 tip1 noc1 SPAC3C7.12 Protein noc1 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MFPLGSVVEVITGERGFVRYAGEVENRKGVYVGLELLPEFAEFGKNRGVVDGREYFKTKNNEKTGIFVPFDKCKLASSISSSPSPKIDGTAASIGMGFPPMSPNLQSSIPRLTNVSSSSNLSMNTISSTALTPTEKILQKRIEDLLYERQNHQQQLEEVLATVDQLQSLVTNFNDQQDEVDELRERITLKEERIQQMRNEASQRRFEFKTTIECLEESSNRAIETYENRIAELEAQLEMYMSGKSEDDLLFSLQQERDYALNQVEILQERVDTLMKQKANSSTANEKLSHMESSSPTLTNASFESPKRGKGSNDLPENHPQRRQTLEFYEIEIEVLREKVEKLQALSDEKDFYISKLEKSLDRNDTTPVPSDEKLSNYAAEKENLVSRISELEHTIEQLTINNERDNERMSPAEFELETTQEVEENDSDSHDDEETWCEVCETNNHSLQECPTVFGSTDEA 66CCDB96146F86D6 for3_schpo Formin-3 for3 SPCC895.05 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MASKMPEGSPPTSRSIQSRNSSYSTSSNERIGTPSTISLSENSDLSKLQSTNDFESREDLSLTSDDNNDPEYVMCYNTVYHQKTKINDKLLSETEQLRKIYPLESRVFPKPTIVKEITNERTKRYTFYDDDAPLTNQHTVLDEATYNRILKRIDFFIEKVVEVPPTYHFLSLLNVQLRIQPIWWMDEFAERNGIESLLSALRNLGHYPERASKTPLESQIIQSLFHCLNSENCRRRYQSSAKCSVPGFNALGTIAETVLSKSLNSARMATFLLKFLCNKKGLSYFKAVIRAFEWLVEQKLSKTRFSAWMHSFNDVITGVRVCADSSPQAIVHMDEFEDTDCLIDYLVATLALIRDLCAAPPDLQLRCALRHELLSAGLQKAIDSLLKWRNRHVRDALQLLIKEHNADARRFRSGSDVNNVDRKCVKKQMNYREESHTPHGNTTRKTSTPVSNNRPTTPEQQAVWDVFQRIYTRFTGSEGSKESFIKLLEYFVTEPDNGKIQKSMQLLTHTLEALEGFKTAKADTNVGLTILSQRLLDKLGTAEEIAEYKTKYNGAMLENKHLKEQVESMLSQLNVGPRDPMQFLKKQLDELKAELNLRDNLLASMQREFETRYRAQIQAYNKLQSQMEHVQNSNEQHLQPGLLNKVSKSFDSVHRRNLSQDSLDAMTEQFSYHVEPNILSGSGIPVRVHTPSKTEDLDESFSGSEISSSPSPLLPDVSDTVEEQQKLLLKSPPPPPPAVIVPTPAPAPIPVPPPAPIMGGPPPPPPPPGVAGAGPPPPPPPPPAVSAGGSRYYAPAPQAEPEPKIDETSLTEEQKIQLEEARKQRKAADDAARAAIEKYTSIPSLRDLHKPTRPLKRVHWQRVDPLPGPNVFTKFCLNFDITAKVFIDNGLLDFLDEKFDNTPREDFVAVEISDQRSSLLPDTVEQNMAIILRSVSNMPVEDLVQKFLVEPDFLPASILYFDRASLASTNAYTDPFIPYSTDYTKKNPKEPTADVNSLSYFEKFFVLFVVNLRHYFQERMKALKFRSTLFGDLEILEVRMKEVIDTSDSIMEDKNFAEFFQVLLIIGNYFNEPYDRASAFSLYMIYRLETLRDSSSALTLMHYFDEIIRTRFPELLQAESTFKKIQSVSGYNIDAMVAGVDGAYDEFCDFQTSLKDGALSKCDQHHPDDKAYDILSEWLPEAKERIRNIKKLKTDMLTKLENTVKYLCEYDSIDKVRNSFFKNLNSFYEMYSIAKAENEERFEKEKRRIMSEDRDKLIRGRQKTSIVAKYRNKRELPEDSDDKQDTASKDKNSLETIDEKMEDASKIEGDAKTGDDNEMEDLDKMEDLEKPDYAEEKDPYITVMSELRSRIQNVPKRTVTVYSDEGVATLEPGAQGDDVVDKAKMILEKMEGHSQLLTSSANPDEEVLRAKLKAAERLQKPAIPRTRRKGHTEPKSAKSLLAELTNGSNASNLVENDRQKQ tea4-tea1-4 Coimmunoprecipitation showing interaction between endogenous tea4 and tea1 541740 N/A 541744 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 false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 541745 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 false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association false false false The tea1-HA coimmunoprecipitation with Tea4-GFP in yeast extracts was determined with an an anti-HA antibody. Figure 1E tea4-tea1-5 Coimmunoprecipitation showing interaction between endogenous tea4 and tea1 541740 N/A 541744 unspecified role unspecified role bait bait 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 false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 541745 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 false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association false false false The tea1-HA coimmunoprecipitation with Tea4-GFP in yeast extracts was determined with an an anti-GFP antibody. Figure 1E tea4-tip1-1 Coimmunoprecipitation showing interaction between endogenous tea4 and tip1 in tea1+ and tea1- cells 541740 N/A 541746 unspecified role unspecified role prey prey schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 541744 unspecified role unspecified role bait bait 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 false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association false false false Tip1 coimmunoprecipitates with tea4 in a tea1-dependent manner. This experiment was performed using cell extracts from tea1 containing and tea4 depleted cells. Figure 2F tea1-tip1-2 Coimmunoprecipitation showing interaction between endogenous tea1 and tip1 in tea4+ and tea4- cells 541740 N/A 541745 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 false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 541746 unspecified role unspecified role prey prey schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association false false false Tip1 coimmunoprecipitates with tea1 in a tea4-indedependent manner. This experiment was performed using cell extracts from tea4 containing and tea4 depleted cells. Figure 2G tea4-for3-3 Coimmunoprecipitation showing interaction between endogenous for3 and tea4 541740 N/A 541747 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 false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 541744 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 false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association false false false Endogenous tagged for3-HA coimmunoprecipitated with Tea4-GFP in yeast extracts; determined with an an anti-HA and anti-myc antibody. Indicating that tea4p and for3p are present in the same complex in vivo. Figure 4A tea1-for3-1 Coimmunoprecipitation showing interaction between endogenous for3 and tea1 in tea4+ and tea4- cells 541740 N/A 541744 unspecified role unspecified role ancillary ancillary schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 541745 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 false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 541747 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 false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast association association false false false Endogenous tagged for3-HA interacted with tea1-HA in yeast extracts from tea4-containing cells and not in tea4-depleted cells; determined with an an anti-HA and anti-myc antibody. Indicating that tea4 is required for tea1 and for3 interaction. Figure 4C tea4-tea1-2 Y2H interaction between tea4 and tea1 fragments 541741 N/A 541745 unspecified role unspecified role bait bait c-terminus region required to bind necessary binding region required to bind deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 541744 unspecified role unspecified role prey prey physical association physical association false false false Shows that the C-terminal half of tea1 was necessary and sufficient to bind to tea4. Figure 1C tea4-tea1-3 Y2H interaction between tea4 fragments and tea1 541741 N/A 541744 unspecified role unspecified role bait bait c-terminus region required to bind necessary binding region required to bind deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 541745 unspecified role unspecified role prey prey physical association physical association false false false The C-terminal third of tea4 mediated the interaction with tea1. Figure 1D tea1-tip1-1 Y2H interaction between tea1 fragments and tip1 541741 N/A 541746 unspecified role unspecified role prey prey N/A 541745 unspecified role unspecified role bait bait n-terminus binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false The N-terminus of tea1 containing kelch repeats, is necessary for the interaction with tip1. Figure 2E tea4-for3-1 Y2H interaction between tea4 and for3 fragments 541741 N/A 541747 unspecified role unspecified role bait bait n-terminus 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 541744 unspecified role unspecified role prey prey physical association physical association false false false Figure 4D tea4-for3-2 Y2H interaction between tea4 fragments and for3 541741 N/A 541744 unspecified role unspecified role bait bait N/A 541747 unspecified role unspecified role prey prey n-terminus 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 No specific domain of tea4p could be defined in for3p binding. Neither the tea4p C-terminal region, which mediates the interaction with tea1p, nor the N-terminal half interacted with for3p reproducibly (Figure 4E), suggesting that multiple parts of the protein may be required for strong interaction. What is clear though is that the SH3 domain of tea4p was dispensable for binding, as shown by deletion or specific point mutation in the SH3 domain. Figure 4E tea4-tea1-6 Pull-down showing interaction between tea4 and tea1 C-terminal fragments. 541742 N/A 541744 unspecified role unspecified role prey prey c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 541745 unspecified role unspecified role bait bait c-terminus mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) physical association physical association false false false Bacterially expressed tea1-MBP bound to tea4-His in in-vitro pull-down experiment. Figure 1F tea1-tea1-1 Pull-down showing tea1 C-terminal dimerization. 541742 N/A 541745 unspecified role unspecified role prey prey c-terminus binding region binding-associated region binding region deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 541745 unspecified role unspecified role bait bait c-terminus binding region binding-associated region binding region deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) physical association physical association false false false Bacterially expressed tea1-MBP bound to tea1-His in in-vitro pull-down experiment as shown in Figures 1F and 4B. Figure 1F tea4-tea1-9 Pull-down showing interaction between tea4 and tea1 C-terminal fragments. 541742 N/A 541744 unspecified role unspecified role bait bait region mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 541745 unspecified role unspecified role prey prey c-terminus binding region binding-associated region binding region deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) physical association physical association false false false Bacterially expressed tea1-His bound to tea4-MBP in in-vitro pull-down experiment. Figure 4B tea4-for3-4 Pull-down showing interaction between tea4 and for3 fragments. 541742 N/A 541747 unspecified role unspecified role bait bait n-terminus binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 541744 unspecified role unspecified role prey prey region mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) physical association physical association false false false Bacterially expressed N-terminus for3-His bound to tea4-MBP in an in-vitro pull-down experiment. Figure 4B tea4-tea1-8 Colocalization showing interaction between tea4 and tea1 in vivo 541743 N/A 541744 unspecified role unspecified role neutral component neutral component region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 541745 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association false false false Tea4-YFP colocalizes precisely with tea1-CFP in vivo. Figure 2B