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/ Nbl1p: a Borealin/Dasra/CSC-1-like protein essential for Aurora/Ipl1 complex function and integrity in Saccharomyces cerevisiae. Nbl1p: a Borealin/Dasra/CSC-1-like protein essential for Aurora/Ipl1 complex function and integrity in Saccharomyces cerevisiae. Mol. Biol. Cell (1059-1524) 2009 imex curation Nakajima Y., Tyers R.G., Wong C.C., Yates J.R., Drubin D.G., Barnes G. I've not done the kinase assay in S1 as there seem to be at least 7 proteins in the tube, so hardly direct. Colocalisation in S2 is a subcellular marker. gbarnes@berkeley.edu 11-Jun-2014: Contacted by IntAct-Help. Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast fluorescence imaging fluorescence microscopy fluorescence imaging tag fluorescence tag visualisation by fluorescence tag fluorescence Nakajima Y., Tyers R.G., Wong C.C., Yates J.R., Drubin D.G., Barnes G. I've not done the kinase assay in S1 as there seem to be at least 7 proteins in the tube, so hardly direct. Colocalisation in S2 is a subcellular marker. gbarnes@berkeley.edu Accepted 2014-MAY-30 by MGT Mol. Biol. Cell (1059-1524) 2009 Only protein-protein interactions imex curation Nbl1p: a Borealin/Dasra/CSC-1-like protein essential for Aurora/Ipl1 complex function and integrity in Saccharomyces cerevisiae. Nbl1p: a Borealin/Dasra/CSC-1-like protein essential for Aurora/Ipl1 complex function and integrity in Saccharomyces cerevisiae. Mol. Biol. Cell (1059-1524) 2009 imex curation Nakajima Y., Tyers R.G., Wong C.C., Yates J.R., Drubin D.G., Barnes G. I've not done the kinase assay in S1 as there seem to be at least 7 proteins in the tube, so hardly direct. Colocalisation in S2 is a subcellular marker. gbarnes@berkeley.edu 11-Jun-2014: Contacted by IntAct-Help. Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast crosslink cross-linking study crosslink sequence tag sequence tag identification sequence tag Nakajima Y., Tyers R.G., Wong C.C., Yates J.R., Drubin D.G., Barnes G. I've not done the kinase assay in S1 as there seem to be at least 7 proteins in the tube, so hardly direct. Colocalisation in S2 is a subcellular marker. Dithiobis[succinimidylpropionate] as cross-linking agent. gbarnes@berkeley.edu Accepted 2014-MAY-30 by MGT Mol. Biol. Cell (1059-1524) 2009 Only protein-protein interactions imex curation Nbl1p: a Borealin/Dasra/CSC-1-like protein essential for Aurora/Ipl1 complex function and integrity in Saccharomyces cerevisiae. Nbl1p: a Borealin/Dasra/CSC-1-like protein essential for Aurora/Ipl1 complex function and integrity in Saccharomyces cerevisiae. Mol. Biol. Cell (1059-1524) 2009 imex curation Nakajima Y., Tyers R.G., Wong C.C., Yates J.R., Drubin D.G., Barnes G. I've not done the kinase assay in S1 as there seem to be at least 7 proteins in the tube, so hardly direct. Colocalisation in S2 is a subcellular marker. gbarnes@berkeley.edu 11-Jun-2014: Contacted by IntAct-Help. Only protein-protein interactions spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection tap tandem affinity purification TAP TAP tap weight by comassie molecular weight estimation by coomasie staining weight by comassie Nakajima Y., Tyers R.G., Wong C.C., Yates J.R., Drubin D.G., Barnes G. I've not done the kinase assay in S1 as there seem to be at least 7 proteins in the tube, so hardly direct. Colocalisation in S2 is a subcellular marker. Tandem affinity purification- according to PMID:11756468 - (Cheeseman et al., -2001) the tag is composed of an S tag, a TEV protease cleavage site, and a ZZ tag. gbarnes@berkeley.edu Accepted 2014-MAY-30 by MGT Mol. Biol. Cell (1059-1524) 2009 Only protein-protein interactions imex curation Nbl1p: a Borealin/Dasra/CSC-1-like protein essential for Aurora/Ipl1 complex function and integrity in Saccharomyces cerevisiae. Nbl1p: a Borealin/Dasra/CSC-1-like protein essential for Aurora/Ipl1 complex function and integrity in Saccharomyces cerevisiae. Mol. Biol. Cell (1059-1524) 2009 imex curation Nakajima Y., Tyers R.G., Wong C.C., Yates J.R., Drubin D.G., Barnes G. I've not done the kinase assay in S1 as there seem to be at least 7 proteins in the tube, so hardly direct. Colocalisation in S2 is a subcellular marker. gbarnes@berkeley.edu 11-Jun-2014: Contacted by IntAct-Help. Only protein-protein interactions yeast Saccharomyces cerevisiae 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 Nakajima Y., Tyers R.G., Wong C.C., Yates J.R., Drubin D.G., Barnes G. I've not done the kinase assay in S1 as there seem to be at least 7 proteins in the tube, so hardly direct. Colocalisation in S2 is a subcellular marker. gbarnes@berkeley.edu Accepted 2014-MAY-30 by MGT Mol. Biol. Cell (1059-1524) 2009 Only protein-protein interactions imex curation Nbl1p: a Borealin/Dasra/CSC-1-like protein essential for Aurora/Ipl1 complex function and integrity in Saccharomyces cerevisiae. Nbl1p: a Borealin/Dasra/CSC-1-like protein essential for Aurora/Ipl1 complex function and integrity in Saccharomyces cerevisiae. Mol. Biol. Cell (1059-1524) 2009 imex curation Nakajima Y., Tyers R.G., Wong C.C., Yates J.R., Drubin D.G., Barnes G. I've not done the kinase assay in S1 as there seem to be at least 7 proteins in the tube, so hardly direct. Colocalisation in S2 is a subcellular marker. gbarnes@berkeley.edu 11-Jun-2014: Contacted by IntAct-Help. Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast tap tandem affinity purification TAP TAP tap western blot western blot Immuno blot Immuno blot Nakajima Y., Tyers R.G., Wong C.C., Yates J.R., Drubin D.G., Barnes G. I've not done the kinase assay in S1 as there seem to be at least 7 proteins in the tube, so hardly direct. Colocalisation in S2 is a subcellular marker. Tandem affinity purification- according to PMID:11756468 - (Cheeseman et al., -2001) the tag is composed of an S tag, a TEV protease cleavage site, and a ZZ tag. gbarnes@berkeley.edu Accepted 2014-MAY-30 by MGT Mol. Biol. Cell (1059-1524) 2009 Only protein-protein interactions imex curation Nbl1p: a Borealin/Dasra/CSC-1-like protein essential for Aurora/Ipl1 complex function and integrity in Saccharomyces cerevisiae. Nbl1p: a Borealin/Dasra/CSC-1-like protein essential for Aurora/Ipl1 complex function and integrity in Saccharomyces cerevisiae. Mol. Biol. Cell (1059-1524) 2009 imex curation Nakajima Y., Tyers R.G., Wong C.C., Yates J.R., Drubin D.G., Barnes G. I've not done the kinase assay in S1 as there seem to be at least 7 proteins in the tube, so hardly direct. Colocalisation in S2 is a subcellular marker. gbarnes@berkeley.edu 11-Jun-2014: Contacted by IntAct-Help. Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast tap tandem affinity purification TAP TAP tap weight silver stain molecular weight estimation by silver staining weight silver stain Nakajima Y., Tyers R.G., Wong C.C., Yates J.R., Drubin D.G., Barnes G. I've not done the kinase assay in S1 as there seem to be at least 7 proteins in the tube, so hardly direct. Colocalisation in S2 is a subcellular marker. Tandem affinity purification- according to PMID:11756468 - (Cheeseman et al., -2001) the tag is composed of an S tag, a TEV protease cleavage site, and a ZZ tag. gbarnes@berkeley.edu Accepted 2014-MAY-30 by MGT Mol. Biol. Cell (1059-1524) 2009 Only protein-protein interactions imex curation nbl1_yeast N-terminal-borealin-like protein NBL1 YHR199C-A protein protein yeast Saccharomyces cerevisiae Baker's yeast MIPALTPEERQKLRSAILHRMQLELETTEKLIENIKEETLKKLNLLQQPDATSAPQSKELIREVLEQEGRRIE 470C704C05F62FBC ipl1_yeast Spindle assembly checkpoint kinase IPL1 YPL209C Aurora kinase Increase-in-ploidy protein 1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MQRNSLVNIKLNANSPSKKTTTRPNTSRINKPWRISHSPQQRNPNSKIPSPVREKLNRLPVNNKKFLDMESSKIPSPIRKATSSKMIHENKKLPKFKSLSLDDFELGKKLGKGKFGKVYCVRHRSTGYICALKVMEKEEIIKYNLQKQFRREVEIQTSLNHPNLTKSYGYFHDEKRVYLLMEYLVNGEMYKLLRLHGPFNDILASDYIYQIANALDYMHKKNIIHRDIKPENILIGFNNVIKLTDFGWSIINPPENRRKTVCGTIDYLSPEMVESREYDHTIDAWALGVLAFELLTGAPPFEEEMKDTTYKRIAALDIKMPSNISQDAQDLILKLLKYDPKDRMRLGDVKMHPWILRNKPFWENKRL 44E3EFDDE1CDB35E bir1_yeast Protein BIR1 J1880 YJR089W BIR1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MDGQIDKMEKRYSMTKLENRLRTFQDGVALEKKKLKWSFKVIPYQAMAKLGFYFDPVIDPKTSKLKKDSVRCCYCHRQTYNVRDCRSKRKDVLETLSNIMRQHLTVTDNKQVCLLIYLRNKLLTDYSFHMGVSDWKNDKYFSNPDDENVINLRKFTFQDNWPHSGSQNEHPLGIEKMVNAGLMRYDSSIEGLGDPSMDKTLMNDTCYCIYCKQLLQGWSINDDPMSRHYKVSQNGNCYFFQTRNRFERIKNDNDSITKNCEVSPTLGENGKREVINTKTASQRQCPLFESPPSSTGPQLDDYNEKTDISVIQHNISVLDGAQGENVKRNSVEEKEQINMENGSTTLEEGNINRDVLADKKEVISTPTAKEIKRPNVQLTQSSSPIKKKRKFKRISPRKIFDEEDSEHSLNNNSANGDNKDKDLVIDFTSHIIKNRDVGRKNAILDDSTDEFSFSNQGHNTFDIPIPTSSHLLKGIDSDNDNVIREDDTGINTDTKGASSKHEKFSVNSEEDLNFSEVKLTGRDSSTNILIRTQIVDQNLGDIDRDKVPNGGSPEVPKTHELIRDNSEKREAQNGEFRHQKDSTVRQSPDILHSNKSGDNSSNITAIPKEEQRRGNSKTSSIPADIHPKPRKNLQEPRSLSISGKVVPTERKLDNINIDLNFSASDFSPSSQSEQSSKSSSVISTPVASPKINLTRSLHAVKELSGLKKETDDGKYFTNKQETIKILEDVSVKNETPNNEMLLFETGTPIASQENKSRKLFDEEFSGKELDIPIDSSTVEIKKVIKPEFEPVPSVARNLVSGTSSYPRNSRLEEQRKETSTSLADNSKKGSSFNEGNNEKEPNAAEWFKIDENRHLVKNYFHDLLKYINNNDATLANDKDGDLAFLIKQMPAEELDMTFNNWVNLKVQSIKREFIDDCDKKLDILRRDYYTATNFIETLEDDNQLIDIAKKMGIL 6B20DC16065B4C39 6B20DC16065B4C39 sli15_yeast Inner centromere protein-related protein SLI15 YBR1206 YBR156C SLI15 protein protein yeast Saccharomyces cerevisiae Baker's yeast MDWAIKAARKKTQRKPGSTRSIIETLDDLNNLTTDAHSEINQRLYESSEWLRNNVYMNTLKYEDKKMEESLISPENTHNKMDVEFPKMKGEYELSNSQNDAAKDVTKTPRNGLHNDKSITPKSLRRKEVTEGMNRFSIHDTNKSPVEPLNSVKVDANESEKSSPWSPYKVEKVLRESSKTSESPINTKRFDNQTWAAKEEMENEPILQALKKAESVKVKPPPNSGIARSQRRSNMFVPLPNKDPLIIQHIPPTKSSGSIPKVRTVKESPIAFKKKSTINSPAIRAVENSDTAGSTKASSVFDRLSSIPTKSFENKISRGNVGHKYSSSSIDLTGSPMKKVSQKFKSINSTDTDMQEALRDIFSVKNKITKNNSPKGKNSRKSSIPRFDKTSLKLTTHKKLAIIAEQKKKSKHSSDVHKTGSRPHSISPTKISVDSSSPSKEVKNYYQSPVRGYLRPTKASISPNKNKNLTTSQTPHRLKIKEKTLRKLSPNIADISKPESRKSKNYRLTNLQLLPPAEAERDDLKKKFDKRLSGIMRSQQEHHRRKQEKQKRMSHLEQDLKKQTSFSNDYKDIRLKESLAPFDNHVRDTINKNTAFSTDNILATINTVDHREIIGNVTPKIASVNDSLPEINTDSEDEASVTLAAWAKSPYLQEQLIRQQDINPQTIFGPIPPLHTDEIFPNPRLNRLKPRQIVPKRS 827AC2AB884BA7D1 827AC2AB884BA7D1 sli15-bir1 5 8 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role prey prey association association Fig. 1C Nbl1p is too small to be seen on a discontinuous gel that is able to resolve Sli15p and Bir1p. nbl1-bir1 5 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey association association Fig. 1C bir1-ipl1 2 7 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component 10 unspecified role unspecified role neutral component neutral component association association No figure ipl1-nbl1 1 7 unspecified role unspecified role neutral component neutral component c-terminal mcherry mcherry fluorescent protein tag mcherry c-term range C-terminal range c-term range c-term range C-terminal range c-term range 8 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Fig. 3C bir1-nbl1 1 9 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 7 unspecified role unspecified role neutral component neutral component c-terminal mcherry mcherry fluorescent protein tag mcherry c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Fig. 3C nbl1-sli15 1 7 unspecified role unspecified role neutral component neutral component c-terminal mcherry mcherry fluorescent protein tag mcherry c-term range C-terminal range c-term range c-term range C-terminal range c-term range 10 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Fig. 3C sli15-bir1-1 3 10 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey physical association physical association Fig. 4D No detectable Bir1 copurified with Sli15-TAP from nbl1-/- cells. Calmodulin was also indicated to be present, but as it was unclear how this was identified, this has not been added to the complex. nbl1-bir1-1 4 7 unspecified role unspecified role bait bait 9 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 physical association physical association Fig. 5 bir1-nbl1-2 4 7 unspecified role unspecified role prey prey 9 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 physical association physical association Fig. 5 bir1-nbl1-3 4 9 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 10 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 58 56 physical association physical association Fig. 5 sli15-bir1-2 4 10 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 9 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 physical association physical association Fig. 5 sli15-nbl1 4 7 unspecified role unspecified role prey prey 10 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 physical association physical association Fig. 5 nbl1-sli15-1 4 7 unspecified role unspecified role bait bait 10 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 physical association physical association Fig. 5 sli15-sli15 4 10 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 10 unspecified role unspecified role bait bait physical association physical association Fig. 5 sli15-sli15-1 4 10 unspecified role unspecified role prey prey 10 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 physical association physical association Fig. 5 sli15-ipl1 4 8 unspecified role unspecified role prey prey 10 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 physical association physical association Fig. 5 ipl1-sli15-2 4 8 unspecified role unspecified role bait bait 10 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 physical association physical association Fig. 5 ipl1-bir1-1 4 8 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey 727-954 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Fig. 5 bir1-ipl1-1 4 9 unspecified role unspecified role bait bait 727-954 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role prey prey physical association physical association Fig. 5 sli15-bir1-3 3 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey association association Fig. 4D Calmodulin was also indicated to be present, but as it was unclear how this was identified, this has not been added to the complex. sli15-bir1-4 6 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig. 4B Interaction observed in wild-type yeast and in a temperature sensitive NBL1 knockdown cells grown at a permissive temperature. sli15-ipl1 6 8 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 4B No detectable Bir1 copurified with Sli15-TAP from nbl1-/- cells grown at a non-permissive temperature.