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 conserved kinase UNC-51 acts with VAB-8 and UNC-14 to regulate axon outgrowth in C. elegans. The conserved kinase UNC-51 acts with VAB-8 and UNC-14 to regulate axon outgrowth in C. elegans. Development (0950-1991) 2004 imex curation Lai T., Garriga G. garriga@berkeley.edu 26-Jun-2011: Contacted by JYOTI. mixed stage C. elegans cDNAs jyoti Only protein-protein interactions in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Lai T., Garriga G. garriga@berkeley.edu Accepted 2011-JUL-2 by JYOTI. 26-Jun-2011: Contacted by JYOTI. mixed stage C. elegans cDNAs Development (0950-1991) 2004 jyoti Only protein-protein interactions imex curation The conserved kinase UNC-51 acts with VAB-8 and UNC-14 to regulate axon outgrowth in C. elegans. The conserved kinase UNC-51 acts with VAB-8 and UNC-14 to regulate axon outgrowth in C. elegans. Development (0950-1991) 2004 imex curation Lai T., Garriga G. garriga@berkeley.edu 26-Jun-2011: Contacted by JYOTI. mixed stage C. elegans cDNAs jyoti Only protein-protein interactions chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Lai T., Garriga G. I dont think these can be phosphorylations as there is no indication as to how the added phosphate groups were detected (only adding phosphatase stopping the reaction) - Fig 7A/B/D/E shows NO radioisotope (as compared to Fig 7C that DOES). 7A/B image legend states:"Western blots were probed with anti-HA antibodies to detect UNC-14." (The Tags used for Figs 7A-C are mud like in clarity...) Supplier web site confirms there is a his tag as well as a myc-tag for VAB-8 in fig 7D. COS cells were used , but the exact cell line is not specified. garriga@berkeley.edu Accepted 2011-JUL-2 by JYOTI. 26-Jun-2011: Contacted by JYOTI. mixed stage C. elegans cDNAs Development (0950-1991) 2004 jyoti Only protein-protein interactions imex curation The conserved kinase UNC-51 acts with VAB-8 and UNC-14 to regulate axon outgrowth in C. elegans. The conserved kinase UNC-51 acts with VAB-8 and UNC-14 to regulate axon outgrowth in C. elegans. Development (0950-1991) 2004 imex curation Lai T., Garriga G. garriga@berkeley.edu 26-Jun-2011: Contacted by JYOTI. mixed stage C. elegans cDNAs jyoti Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Lai T., Garriga G. I think the tags used in this expt are the same as the previous one - Lai-2004b-4 - but it is not very clear if this is so. wild-type UNC-51 was purified from COS cells, but the exact COS cell line used is not stated. garriga@berkeley.edu Accepted 2011-JUL-2 by JYOTI. 26-Jun-2011: Contacted by JYOTI. mixed stage C. elegans cDNAs Development (0950-1991) 2004 jyoti Only protein-protein interactions imex curation The conserved kinase UNC-51 acts with VAB-8 and UNC-14 to regulate axon outgrowth in C. elegans. The conserved kinase UNC-51 acts with VAB-8 and UNC-14 to regulate axon outgrowth in C. elegans. Development (0950-1991) 2004 imex curation Lai T., Garriga G. garriga@berkeley.edu 26-Jun-2011: Contacted by JYOTI. mixed stage C. elegans cDNAs jyoti 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 Lai T., Garriga G. garriga@berkeley.edu Accepted 2011-JUL-2 by JYOTI. 26-Jun-2011: Contacted by JYOTI. mixed stage C. elegans cDNAs Development (0950-1991) 2004 jyoti Only protein-protein interactions imex curation The conserved kinase UNC-51 acts with VAB-8 and UNC-14 to regulate axon outgrowth in C. elegans. The conserved kinase UNC-51 acts with VAB-8 and UNC-14 to regulate axon outgrowth in C. elegans. Development (0950-1991) 2004 imex curation Lai T., Garriga G. garriga@berkeley.edu 26-Jun-2011: Contacted by JYOTI. mixed stage C. elegans cDNAs jyoti 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Lai T., Garriga G. garriga@berkeley.edu Accepted 2011-JUL-2 by JYOTI. 26-Jun-2011: Contacted by JYOTI. mixed stage C. elegans cDNAs Development (0950-1991) 2004 jyoti Only protein-protein interactions imex curation unc51_caeel Serine/threonine-protein kinase unc-51 unc-51 Y60A3A.1 Uncoordinated protein 51 protein protein caeel Caenorhabditis elegans MEQFDGFEYSKRDLLGHGAFAIVYRGRYVDRTDVPVAIKAIAKKNISKSKNLLTKEIKILKELSSLKHENLVGLLKCTETPTHVYLVMEFCNGGDLADYLQQKTTLNEDTIQHFVVQIAHALEAINKKGIVHRDLKPQNILLCNNSRTQNPHFTDIVIKLADFGFARFLNDGVMAATLCGSPMYMAPEVIMSMQYDAKADLWSIGTILFQCLTGKAPFVAQTPPQLKAYYEKTRELRPNIPEWCSPNLRDLLLRLLKRNAKDRISFEDFFNHPFLTSPLLPSPSKRILESARSPLLANRRIITPQSSLPVPKRAGSTKLDSPTPVRRIGESPRVQRRVITPGMPSPVPGAPMQESTDFTFLPPRQESSPVKQVQVHTNVSPSLTTCKPVPVPSQRLTYQKMEERLAAARKTAVPSSSSPTGSAVSAQHQHQHQQQQEPASSPVVQRIERPDQLPRRTTLQDPNAHDIERMTMPNPTFVVCGSSTKPSPNNANRVRRSTITSPADTQDMVAADQMLSNLDPTTTTTTIPKSATTANIQGIPRGARDRSVTSPPQPTIHENEPLDNAKYQQTDVNNSPTAPTEPFIIKNQTTCSTSSTSSSVVEEEEAMSLPFASGSHLAAGFKKTPAEVPMDHGALPPALDQEIVLGEEHKQILAKLRFVAELVDTLIHVAEQKDNPLASAMASRRQLLTTGTSTTNTSSPYRRAEQLVVYVRALHMLSSALLLAQTNVANRVLHPSVAVQQVLNQLNDKYHQCLVRSQELASLGLPGQDPAMAVISAERIMYRHAIELCQAAALDELFGNPQLCSQRYQTAYMMLHTLAEQVNCDQDKTVLTRYKVAVEKRLRILERQGFVAAVNT 721F6F6FB0399F6E g5ecq1_caeel K10D3.2 unc-14 CELE_K10D3.2 protein protein caeel Caenorhabditis elegans MVELCELHMMPEEDQWPDDIDARIAALNFCGEDSMLFESVDPSVSTDSLDSQQFRERCQMKKEDFQLAFADSGHWQSGINDNLTTWGRIRTSEPLDERTASAPDVWNVKRSDSARSPNRPNSLIANFVSGDATRFVDVNDNEIREANEEIIRKDRWRRDSARRCSSGGQNQKRTFADILEKNVTAPTSMAITSSDNEKPPKLDFLAMHHEMPSLCESFTASFRDAIIKMQKCEPLPSITSTNDFPLFFQEDSPDSGLGCSGPSHIEDWQSLSVLLPKHVAEACSFFKSNTQLLTSSTSKTAPQTSTNIVSNCIDRRISGISASANEACRTCYRVRRRIHPPVWAQTAQSKTVLCDCASTPTDTNFSFAPTTSTTRHQLRAKELSIVGLPIYAAKRTLVENVVEGVAAISRGDGSDLLVIAMRCLIEDGLQENVSAWTMIQTVTSKGPATKDVHSIVKQLEECSKTDNVKVEIFFEELIRENSLDCWLCYIVLKEKVLKTLYSENAFLLSASSEYRTLLWRMVDSLSLLPVIEARSDSVHQQFKSMQQWGGASRIASDSRVPKSSSFPARLSTAPSRRSRIPLSTSRISISSTTSTPRSARSPSTTSRIRVASIMGDFTLANFSLSDGEKVSVLSTRGGLARCVRLTTSHSKINNGVIPIEHLLFQ 3B2BFA9D634DFCA4 ki26l_caeel Kinesin-like protein vab-8 vab-8 Variable abnormal morphology protein 8 K12F2.2 protein protein caeel Caenorhabditis elegans MEACSSKTSLLLHSPLRTIPKLRLCASISSEDVAHGRCSLTDQHLQIEGKNYSKTTFDHIFRTDATQDDMYTAFLSDTINSVFAGNDATVLAMGAKTNGKDERLYGNSVSRNGLVQMAITQLMNALDDNKDSEERIQVRMSAIMVSQNESSIVDLLSPFNPDPRHRVVKIVDDARTGVFIDNESEIRVETIDQALFYLNTAVDHRMIQDEHTHRTSHVFISLSLYSYKMGDKMQGGRRRLCFLDMGIGERNSTNGGMTMPALGSILLAMVQRNKHIPSRDSSVCQLIRCALSTSRFTTFVFSFGAKSDDNENIAHLACKIARTRAKSMVGHGRKSSGTMSTGTMESNSSSCGTTTITPGGTPRTQRRFELESGSELSAAETVIFLGPNSSRTASPASTTMPFTPTSIRPLHRTTRNHSGVEALSKPLSVETKSSPTHNCHDGCIHSIPPMLRGHTPFLSASLKLYDELCSPPSSSRASPAPPAAFGGKTSEKREDFGIMIAQPSIPLMKAKSKYNLDDGKMKQIMQWMETSEAPPILFSSPCYENSATSVEELRECVGILSHPLEDIIEQEEESMRTSTATTGGSKKDHPLRILSKQDLNVEPEIKDKQEENELELVMAASLSSMRSHDILAKLEAMRNAQNGNSVQNIGNSQSNTDMDVSEMDVYRRASHLEEYAMQRVREIEENKLKNKKKIKLGLNCCQQQSMISSGSTVVDWSQIERKKEKEREVHEEEMRKEMLRERRAKLKITELEIKRERNLIDKELDDKKGIANSIARQLQHFSLSPCRGGRTHRSVSTHRIDPPNASLPSTPTMSHKKVIGGSLAKLSASGASGSTGAPPSPALGYHQSLPRHSKLPTSVNGRRASAERERKSNKASRNSCSKERKISGSKEELQWRSPYAQMTSPKSYGGPGTSSSGRGSSAPGSDFETPVVSTTEKSANGTIPRSKRQSYSASSGYESANDYHIYSTTNKKPHILDKKRNEEKLSLVRQADEIRHRQWQLKKELEEAKRAIGQEDDAKMIANSSDQRLNGLSRTTMIDAMLQENRILEKRLVACRNHSMLVTTFI A848740A95EC6EF0 vab_8-unc_51 5 8 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association vab_8-unc_51-1 4 6 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 physical association physical association Fig 1A unc_51-vab_8 1 6 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 8 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 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 Fig 1B/C unc_51-unc_14-2 2 7 unspecified role unspecified role prey prey 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 region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role bait bait 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 physical association physical association Fig 7A/B unc_14-unc_51-1 3 7 enzyme target enzyme target substrate substrate neutral component neutral component 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 region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 enzyme enzyme 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 caere Caenorhabditis remanei Caenorhabditis vulgaris phosphorylation phosphorylation reaction phosphorylation Fig 7C UNC-14 is phosphorylated by UNC-51 unc_51-vab_8-1 2 8 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role bait bait 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 physical association physical association Fig 7D