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/ bamps-2004-1 Mcs2 and a novel CAK subunit Pmh1 associate with Skp1 in fission yeast. schpm Schizosaccharomyces pombe (fission yeast) Fission 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 Bamps S., Westerling T., Pihlak A., Tafforeau L., Vandenhaute J., Makela TP., Hermand D. Damien.Hermand@fundp.ac.be Biochem. Biophys. Res. Commun. (0006-291X) 2004 Only protein-protein interactions imex curation bamps-2004-3 Mcs2 and a novel CAK subunit Pmh1 associate with Skp1 in fission yeast. in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip predetermined predetermined participant predetermined Bamps S., Westerling T., Pihlak A., Tafforeau L., Vandenhaute J., Makela TP., Hermand D. Soluble protein extracts from strains expressing either Mcs2 or Pmh1 with C-terminal TAP tags at their endogenous locus (Mcs2-TAP and Pmh1-TAP, respectively) and myc-Skp1 were used in TAP-tag affinity precipitation (by IgG�Sepharose). As the authors were unable to generate a tagged version of Skp1 at the genomic locus we used an ectopically expressed N-terminal myc-tagged Skp1 (myc-Skp1). Damien.Hermand@fundp.ac.be Biochem. Biophys. Res. Commun. (0006-291X) 2004 Only protein-protein interactions imex curation bamps-2004-2 Mcs2 and a novel CAK subunit Pmh1 associate with Skp1 in fission yeast. in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Bamps S., Westerling T., Pihlak A., Tafforeau L., Vandenhaute J., Makela TP., Hermand D. Damien.Hermand@fundp.ac.be Biochem. Biophys. Res. Commun. (0006-291X) 2004 Only protein-protein interactions imex curation cgm2_schpo Cyclin mcs2 Mitotic catastrophe suppressor 2 mcs2 SPBP16F5.02 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSADKFRDSTHYRDWIFTEEDLSKTRAKVNEKFTNIVRERMLEELSLQNKEASLEVLPPTLTVEEELELVNYYSFQLNALSSALSLPTHIRSTAILFFKRFYLINSVMEYSPKIISFTSLFLATKCNDHYISIEQFCKNMPKTTPEEVLEYEFNVCQSLKWDLYVWLPFRPLQGFLLDCQTVLPKVAVEKFYECHDLSKKFLIETLHSDIYFLHSPSIIALGAIYHTNPTICLQYIEAKKIPELQPLIISISANLKATKKFKIEKKKAQDYGRKLYFCMNPLRNKSSALYLKRKAEEESTNNNKWAKKFSTSSNVLDKNPFE skp1_schpo Suppressor of kinetochore protein 1 skp1 psh1 sph1 SPBC409.05 P19/Skp1 homolog protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSKIKLISSDNEEFVVDQLIAERSMLIKNMLEDVGEINVPIPLPNVSSNVLRKVLEWCEHHKNDLYSGTEEESDIRLKKSTDIDEWDRKFMAVDQEMLFEIVLASNYLDIKPLLDTGCKTVANMIRGKSPEDIRKTFNIPNDFTPEEEEQIRKENEWAEDR crk1_schpo Serine/threonine-protein kinase crk1 Mitotic catastrophe suppressor 6 crk1 mcs6 mop1 SPBC19F8.07 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MDIEKSDKWTYVKERKVGEGTYAVVFLGRQKETNRRVAIKKIKVGQFKDGIDISALREIKFLRESRHDNVIELVDVFSTKSNLNIILEFLDSDLEMLIKDKFIVFQPAHIKSWMVMLLRGLHHIHSRFILHRDLKPNNLLISSDGVLKLADFGLSRDFGTPSHMSHQVITRWYRPPELFMGCRSYGTGVDMWSVGCIFAELMLRTPYLPGESDLDQLNVIFRALGTPEPEVIKSMQQLPNYVEMKHIPPPNGGMEALFSAAGHEEIDLLKMMLDYNPYRRPTAQQALEHHYFSALPKPTHPSLLPRKGGEEGIKHVSSDLQRQNNFPMRANIKFV tfb3_schpo RNA polymerase II transcription factor B subunit 3 RNA polymerase II transcription factor B 38 kDa subunit RNA polymerase II transcription factor B p38 subunit CDK-activating kinase assembly factor MAT1 homolog RING finger protein pmh1 pmh1 mcr1 tfb3 SPBC776.18c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MDDEGARKVDEKCPLCQADRYLNPNMKLLINPECYHKMCESCVDRIFTTGPAQCPTPGCNKILRKAKFREQTFEDAQIEREVDVRKRISRIFNKGQQEFDSLQAYNDYLEEVEILTFNLIYKIDVEETEEKVKQYEKQNRDSIAANSARAAAEARILAQNEILLKRQKQEAREAAIREHQKEKERREQVEQQIIFDLATSGKDPNKIIQLSDSLKKQQENIASSVSNISRSSSILLSDVQQVAEDTTPFSPLAGEKDGSKYFSYSKNTYQDLYLEKVSHEPGRKCGFRIEDCHLRCLYEAFSGIDYDLESLKKLEVAS mcs2-skp1 Y2H interaction between mcs2 and skp1 in schpo 570032 N/A 570035 unspecified role unspecified role bait bait N/A 570036 unspecified role unspecified role prey prey c-terminal 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 The interaction between lexA-Mcs2 and B42-Skp1 was measured via activation of LEU2 and lacZ reporters in Y2H. Figure 1A mcs2-skp1-1 Y2H interaction between mcs2 and skp1 in schpo 570032 N/A 570035 unspecified role unspecified role bait bait N/A 570036 unspecified role unspecified role prey prey Q9Y709:p.Ile139Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false The interaction between lexA-Mcs2 and B42-Skp1 was confirmed via quantitative beta-galactosidase assay in Y2H. Additionally it showed that the interaction with Mcs2 requires the correct amino acid at position 172 since the mutation I172R abolishes this interaction. Figure 1B mcs2-mcs6 Y2H interaction between mcs2 and mcs6 in schpo 570032 N/A 570037 unspecified role unspecified role prey prey N/A 570035 unspecified role unspecified role bait bait physical association physical association false false false The interaction between lexA-Mcs2 and its cognate CDK B42-Mcs6 was confirmed via quantitative beta-galactosidase assay in Y2H. Figure 1B skp1-pmh1 Y2H interaction between skp1 and pmh1 in schpo 570032 N/A 570038 unspecified role unspecified role prey prey N/A 570036 unspecified role unspecified role bait bait physical association physical association false false false Skp1 was used as bait to pick up and identify a novel full-length cDNA encoding pmh1 in fission yeast. Figure 1C mcs2-skp1-2 CoIP showing interaction between mcs2 and skp1 in schpo 570033 N/A 570035 unspecified role unspecified role prey prey c-terminal tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 570036 unspecified role unspecified role bait bait n-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association false false false Figure 3A myc-Skp1 was able to copurify Mcs2-TAP. skp1-pmh1-1 CoIP showing interaction between skp1 and pmh1 in schpo 570033 N/A 570038 unspecified role unspecified role prey prey c-terminal tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 570036 unspecified role unspecified role bait bait n-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association false false false myc-Skp1 copurified Pmh1-TAP. Figure 3B mcs2-mcs6-1 CoIp (anti-bait) showing interaction between mcs2, mcs6 and tbf3 in schpo 570034 N/A 570038 unspecified role unspecified role prey prey 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 false trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. N/A 570035 unspecified role unspecified role prey prey trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. N/A 570037 unspecified role unspecified role bait bait trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. association association false false false Figure 2B