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/ Two complexes of spindle checkpoint proteins containing Cdc20 and Mad2 assemble during mitosis independently of the kinetochore in Saccharomyces cerevisiae. Two complexes of spindle checkpoint proteins containing Cdc20 and Mad2 assemble during mitosis independently of the kinetochore in Saccharomyces cerevisiae. Eukaryotic Cell (1535-9778) 2005 imex curation Poddar A., Stukenberg PT., Burke DJ. Superose 6 column djb6t@virginia.edu 07-Aug-2007: Contacted by JYOTI. Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column anti tag western anti tag western blot anti tag western Poddar A., Stukenberg PT., Burke DJ. Superose 6 column djb6t@virginia.edu Accepted 2007-AUG-5 by JYOTI. Eukaryotic Cell (1535-9778) 2005 Only protein-protein interactions imex curation Two complexes of spindle checkpoint proteins containing Cdc20 and Mad2 assemble during mitosis independently of the kinetochore in Saccharomyces cerevisiae. Two complexes of spindle checkpoint proteins containing Cdc20 and Mad2 assemble during mitosis independently of the kinetochore in Saccharomyces cerevisiae. Eukaryotic Cell (1535-9778) 2005 imex curation Poddar A., Stukenberg PT., Burke DJ. Superose 6 column djb6t@virginia.edu 07-Aug-2007: Contacted by JYOTI. Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Poddar A., Stukenberg PT., Burke DJ. Superose 6 column djb6t@virginia.edu Accepted 2007-AUG-5 by JYOTI. 07-Aug-2007: Contacted by JYOTI. Eukaryotic Cell (1535-9778) 2005 Only protein-protein interactions imex curation cdc20_yeast APC/C activator protein CDC20 YGL116W CDC20 Cell division control protein 20 protein protein yeast Saccharomyces cerevisiae Baker's yeast MPESSRDKGNAAISGNRSVLSIASPTKLNILSSDWSRNQGKVSKNSLKRSSSLNIRNSKRPSLQASANSIYSRPKITIGAPPLIRRDSSFFKDEFDAKKDKATFSAYSSRSYPTIGSESVVSQTSLSQPTTSREVDEQFTVAADRYIPILQGASQNKVDPETLHEALPPPNASPISHLRAQTKIVFKQNVAEACGLDMNKRILQYMPEPPKCSSLRQKSYIMKKRTHYSYQQEQKIPDLIKLRKINTNPERILDAPGFQDDFYLNLLSWSKKNVLAIALDTALYLWNATTGDVSLLTDFENTTICSVTWSDDDCHISIGKEDGNTEIWDVETMSLIRTMRSGLGVRIGSLSWLDTLIATGSRSGEIQINDVRIKQHIVSTWAEHTGEVCGLSYKSDGLQLASGGNDNTVMIWDTRTSLPQFSKKTHTAAVKALSWCPYSPNILASGGGQTDKHIHFWNSITGARVGSINTGSQVSSLHWGQSHTSTNGGMMNKEIVATGGNPENAISVYNYETKFKVAEVVHAHEARICCSQLSPDGTTLATVGGDENLKFYKIFDPRCTGRSREDGLMDGMLGLIGKEGCRTNDKENRSKNSSEIHTRRPSSTSQYLIR CCE7CD149C1F5ACF CCE7CD149C1F5ACF bub3_yeast Cell cycle arrest protein BUB3 OR26.16 YOR026W BUB3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MQIVQIEQAPKDYISDIKIIPSKSLLLITSWDGSLTVYKFDIQAKNVDLLQSLRYKHPLLCCNFIDNTDLQIYVGTVQGEILKVDLIGSPSFQALTNNEANLGICRICKYGDDKLIAASWDGLIEVIDPRNYGDGVIAVKNLNSNNTKVKNKIFTMDTNSSRLIVGMNNSQVQWFRLPLCEDDNGTIEESGLKYQIRDVALLPKEQEGYACSSIDGRVAVEFFDDQGDDYNSSKRFAFRCHRLNLKDTNLAYPVNSIEFSPRHKFLYTAGSDGIISCWNLQTRKKIKNFAKFNEDSVVKIACSDNILCLATSDDTFKTNAAIDQTIELNASSIYIIFDYEN 0BDFB8697935BCEB 0BDFB8697935BCEB cdc27_yeast Anaphase-promoting complex subunit CDC27 APC3 CDC27 SNB1 YBL084C YBL0718 Cell division control protein 27 Anaphase-promoting complex subunit 3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MAVNPELAPFTLSRGIPSFDDQALSTIIQLQDCIQQAIQQLNYSTAEFLAELLYAECSILDKSSVYWSDAVYLYALSLFLNKSYHTAFQISKEFKEYHLGIAYIFGRCALQLSQGVNEAILTLLSIINVFSSNSSNTRINMVLNSNLVHIPDLATLNCLLGNLYMKLDHSKEGAFYHSEALAINPYLWESYEAICKMRATVDLKRVFFDIAGKKSNSHNNNAASSFPSTSLSHFEPRSQPSLYSKTNKNGNNNINNNVNTLFQSSNSPPSTSASSFSSIQHFSRSQQQQANTSIRTCQNKNTQTPKNPAINSKTSSALPNNISMNLVSPSSKQPTISSLAKVYNRNKLLTTPPSKLLNNDRNHQNNNNNNNNNNNNNNNNNNNNNNNNIINKTTFKTPRNLYSSTGRLTTSKKNPRSLIISNSILTSDYSITLPEIMYNFALILRSSSQYNSFKAIRLFESQIPSHIKDTMPWCLVQLGKLHFEIINYDMSLKYFNRLKDLQPARVKDMEIFSTLLWHLHDKVKSSNLANGLMDTMPNKPETWCCIGNLLSLQKDHDAAIKAFEKATQLDPNFAYAYTLQGHEHSSNDSSDSAKTCYRKALACDPQHYNAYYGLGTSAMKLGQYEEALLYFEKARSINPVNVVLICCCGGSLEKLGYKEKALQYYELACHLQPTSSLSKYKMGQLLYSMTRYNVALQTFEELVKLVPDDATAHYLLGQTYRIVGRKKDAIKELTVAMNLDPKGNQVIIDELQKCHMQE A1ECA067AF8784E3 A1ECA067AF8784E3 mad3_yeast Spindle assembly checkpoint component MAD3 MAD3 YJL013C J1341 Mitotic MAD3 protein protein protein yeast Saccharomyces cerevisiae Baker's yeast MKAYAKKRISYMPSSPSQNVINFEEIETQKENILPLKEGRSAAALSKAIHQPLVEINQVKSSFEQRLIDELPALSDPITLYLEYIKWLNNAYPQGGNSKQSGMLTLLERCLSHLKDLERYRNDVRFLKIWFWYIELFTRNSFMESRDIFMYMLRNGIGSELASFYEEFTNLLIQKEKFQYAVKILQLGIKNKARPNKVLEDRLNHLLRELGENNIQLGNEISMDSLESTVLGKTRSEFVNRLELANQNGTSSDVNLTKNNVFVDGEESDVELFETPNRGVYRDGWENFDLKAERNKENNLRISLLEANTNLGELKQHEMLSQKKRPYDEKLPIFRDSIGRSDPVYQMINTKDQKPEKIDCNFKLIYCEDEESKGGRLEFSLEEVLAISRNVYKRVRTNRKHPREANLGQEESANQKEAEAQSKRPKISRKALVSKSLTPSNQGRMFSGEEYINCPMTPKGRSTETSDIISAVKPRQLTPILEMRESNSFSQSKNSEIISDDDKSSSSFISYPPQR 36550249D81BB6D1 36550249D81BB6D1 mad2_yeast Mitotic spindle checkpoint component MAD2 MAD2 J1256 YJL030W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSQSISLKGSTRTVTEFFEYSINSILYQRGVYPAEDFVTVKKYDLTLLKTHDDELKDYIRKILLQVHRWLLGGKCNQLVLCIVDKDEGEVVERWSFNVQHISGNSNGQDDVVDLNTTQSQIRALIRQITSSVTFLPELTKEGGYTFTVLAYTDADAKVPLEWADSNSKEIPDGEVVQFKTFSTNDHKVGAQVSYKY EFE59916C5720644 mad3-bub3-2 1 5 unspecified role unspecified role neutral component neutral component c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 3 unspecified role unspecified role neutral component neutral component c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 4 unspecified role unspecified role neutral component neutral component c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role neutral component neutral component c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Nocodazole - activates the spindle checkpoint. MW = 670 kDa The fractionation profiles, Fig. 1D, demonstrate that Bub3, Mad3, Cdc20, and Cdc27 comigrated as a large multiprotein complex (670 kDa) when extracts were prepared from cells grown in the presence of nocodazole to activate the spindle checkpoint. 1D mad2-bub3-2 1 4 unspecified role unspecified role neutral component neutral component c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 3 unspecified role unspecified role neutral component neutral component c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope 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 5 unspecified role unspecified role neutral component neutral component c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role neutral component neutral component c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Nocodazole - activates the spindle checkpoint. MW = 670 kDa In one of the peaks, Mad2 cofractionated with the Bub3, Mad3, Cdc20, and Cdc27 complex, 1D mad2-cdc20-1 2 7 unspecified role unspecified role prey prey 3 unspecified role unspecified role bait bait c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association A greater amount of Mad2 coimmunoprecipitated with Cdc20-13Myc than with either Bub3-13Myc or Mad3-13Myc. 1F, 2D Mad2 and Cdc20 coimmunoprecipitate from Mad3-ProteinA-depleted extracts showing that Mad2 and Cdc20 form a separate complex that does not contain Mad3. mad2-bub3-4 2 4 unspecified role unspecified role bait bait c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 7 unspecified role unspecified role prey prey physical association physical association A greater amount of Mad2 coimmunoprecipitated with Cdc20-13Myc than with either Bub3-13Myc or Mad3-13Myc. 1F mad2-mad3-1 2 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association A greater amount of Mad2 coimmunoprecipitated with Cdc20-13Myc than with either Bub3-13Myc or Mad3-13Myc. 1F mad2-mad3-2 2 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait c-terminal protein a tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association and a small fraction of Mad2 copurified with Mad3-PrA (Fig. 2C, lanes 5 and 6), consistent with our previous observation. 2C cdc20-mad3-1 2 3 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait c-terminal protein a tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association A small amount of Cdc20 copurified with Mad3-PrA (Fig. 2D, lanes 4 to 6) demonstrating that this complex contains Mad2, Mad3-PrA, and Cdc20-13Myc and therefore is MCC (37). 2D