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/ Cyclin B proteolysis and the cyclin-dependent kinase inhibitor rum1p are required for pheromone-induced G1 arrest in fission yeast. Cyclin B proteolysis and the cyclin-dependent kinase inhibitor rum1p are required for pheromone-induced G1 arrest in fission yeast. Mol. Biol. Cell (1059-1524) 1998 imex curation Stern B., Nurse P. Only protein-protein interactions schpm Schizosaccharomyces pombe (fission yeast) Fission yeast anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Stern B., Nurse P. Accepted 2007-APR-28 by DAVET. Mol. Biol. Cell (1059-1524) 1998 Only protein-protein interactions imex curation Cyclin B proteolysis and the cyclin-dependent kinase inhibitor rum1p are required for pheromone-induced G1 arrest in fission yeast. Cyclin B proteolysis and the cyclin-dependent kinase inhibitor rum1p are required for pheromone-induced G1 arrest in fission yeast. Mol. Biol. Cell (1059-1524) 1998 imex curation Stern B., Nurse P. Only protein-protein interactions schpm Schizosaccharomyces pombe (fission yeast) Fission yeast anti tag coip anti tag coimmunoprecipitation anti tag coip predetermined predetermined participant predetermined Stern B., Nurse P. Accepted 2007-APR-28 by DAVET. Mol. Biol. Cell (1059-1524) 1998 Only protein-protein interactions imex curation cg23_schpo G2/mitotic-specific cyclin cdc13 cdc13 SPBC582.03 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MTTRRLTRQHLLANTLGNNDENHPSNHIARAKSSLHSSENSLVNGKKATVSSTNVPKKRHALDDVSNFHNKEGVPLASKNTNVRHTTASVSTRRALEEKSIIPATDDEPASKKRRQPSVFNSSVPSLPQHLSTKSHSVSTHGVDAFHKDQATIPKKLKKDVDERVVSKDIPKLHRDSVESPESQDWDDLDAEDWADPLMVSEYVVDIFEYLNELEIETMPSPTYMDRQKELAWKMRGILTDWLIEVHSRFRLLPETLFLAVNIIDRFLSLRVCSLNKLQLVGIAALFIASKYEEVMCPSVQNFVYMADGGYDEEEILQAERYILRVLEFNLAYPNPMNFLRRISKADFYDIQTRTVAKYLVEIGLLDHKLLPYPPSQQCAAAMYLAREMLGRGPWNRNLVHYSGYEEYQLISVVKKMINYLQKPVQHEAFFKKYASKKFMKASLFVRDWIKKNSIPLGDDADEDYTFHKQKRIQHDMKDEEW B2C267CF57F5FD36 B2C267CF57F5FD36 cg22_schpo G2/mitotic-specific cyclin cig2 cig2 cyc17 SPAPB2B4.03 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MALYSISKPVGSKINKHSYQDENTLVGKQALSKGTEKTKLSTNFEINLPRRTVLSDVSNVGKNNADEKDTKKAKRSFDESNLSTNEEADKPVESKFVKKLKVYSKNADPSVETLQKDRVSNVDDHLSSNPLMAEEYAPEIFEYIRKLDLKCLPNPKYMDQQKELTWKMREILNEWLVEIHSNFCLMPETLYLAVNIIDRFLSRRSCSLSKFQLTGITALLIASKYEEVMCPSIQNFVYMTDGAFTVEDVCVAERYMLNVLNFDLSYPSPLNFLRKISQAEGYDAQTRTLGKYLTEIYLFDHDLLRYPMSKIAAAAMYLSRRLLRRGPWTPKLVESSGGYEEHELKEIAYIMLHYHNKPLEHKAFFQKYSSKRFLKASIFVHQLVRQRYSVNRTDDDDLQSEPSSSLTNDGH 6F255D2E26C6F8B3 rum1_schpo Cyclin-dependent kinase inhibitor rum1 rum1 SPBC32F12.09 p25-rum1 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MEPSTPPMRGLCTPSTPESPGSFKGVIDASLEGNSSIMIDEIPESDLPAPQVSTFPPTPAKTPKKQLLPNLMLQDRSNSLERCMEEDREHNPFLSSSDNQLLSRKKRKPTPPPSDGLYYVFRGKRIKKSFRPGTDLSTFKPKLLFADSAPSSSSDNPTSSVDLNDYSQIGILPPNLNSIGNKMFSLKSRVPSSSSGSFVAPPPQMRLPAYSSPQKSRSNTKDENRHNLLR C5199FE345F7484A cdc13-rum1 1 3 unspecified role unspecified role bait bait schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 5 unspecified role unspecified role prey prey n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag anti tag western anti tag western blot anti tag western n-term range n-terminal range n-term range n-term range n-terminal range n-term range schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association Figure 3B. Given these results, we used a rum1-HAcyr1Dsxa2D strain to test whether rum1p physically associated with cdc13p after pheromone addition. Immunoprecipitations of cdc13p and cig2p before and after addition of pheromone were analyzed by Western blotting with anti-HA antibodies. rum1p was found associated with cdc13p, and this association was increased after 3 h in P-factor (Figure 3B). rum1p was also found associated with cig2p, although the amount detected was lower (Figure 3B). Shows that rum1 physically associated with cdc13 after pheromone addition. cig2-rum1 1 4 unspecified role unspecified role bait bait schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 5 unspecified role unspecified role prey prey n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag anti tag western anti tag western blot anti tag western n-term range n-terminal range n-term range n-term range n-terminal range n-term range schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association Figure 3B. Given these results, we used a rum1-HAcyr1Dsxa2D strain to test whether rum1p physically associated with cdc13p after pheromone addition. Immunoprecipitations of cdc13p and cig2p before and after addition of pheromone were analyzed by Western blotting with anti-HA antibodies. rum1p was found associated with cdc13p, and this association was increased after 3 h in P-factor (Figure 3B). rum1p was also found associated with cig2p, although the amount detected was lower (Figure 3B). Shows that rum1 weakly associated with cig2 after pheromone addition. cig2-rum1-1 2 4 unspecified role unspecified role prey prey schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 5 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag anti tag western anti tag western blot anti tag western n-term range n-terminal range n-term range n-term range n-terminal range n-term range schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association Figure 3C. In a reciprocal experiment, rum1HAp immunoprecipitation was found to coprecipitate 2 pmol of cig2p and 6 pmol of cdc13p (Figure 3C). Similar results were obtained by immunoprecipitations of rum1p from a cig2-HAcyr1Dsxa2D strain (our unpublished results). The different amounts of cig2p and cdc13p in rum1p precipitations are similar to the different cyclin B concentrations in the cell, because the level of cdc13p is about three times that of cig2p (Figure 3C and our unpublished results). Shows that rum1 can associates with 2 pmol of cig2. cdc13-rum1-1 2 3 unspecified role unspecified role prey prey schpm Schizosaccharomyces pombe (fission yeast) Fission yeast 5 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag anti tag western anti tag western blot anti tag western n-term range n-terminal range n-term range n-term range n-terminal range n-term range schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association Figure 3C. In a reciprocal experiment, rum1HAp immunoprecipitation was found to coprecipitate 2 pmol of cig2p and 6 pmol of cdc13p (Figure 3C). Similar results were obtained by immunoprecipitations of rum1p from a cig2-HAcyr1Dsxa2D strain (our unpublished results). The different amounts of cig2p and cdc13p in rum1p precipitations are similar to the different cyclin B concentrations in the cell, because the level of cdc13p is about three times that of cig2p (Figure 3C and our unpublished results). The increased amount of cdc13p associated with rum1p is consistent with rum1p having an effect on the cdc13passociated cdc2p protein kinase. Shows that rum1 can associate with 6 pmol of cdc13.