UniProt UniProt http://www.uniprot.org le_masson-2002-1 Spc24 interacts with Mps2 and is required for chromosome segregation, but is not implicated in spindle pole body duplication. 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 Le Masson I., Saveanu C., Chevalier A., Namane A., Gobin R., Fromont-Racine M., Jacquier A., Mann C. mann@jonas.saclay.cea.fr FRYL genomic DNA library. Mol. Microbiol. (0950-382X) 2002 le_masson-2002-5 Spc24 interacts with Mps2 and is required for chromosome segregation, but is not implicated in spindle pole body duplication. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast tap tandem affinity purification TAP TAP tap ms participant Identification by mass spectrometry ms participant Le Masson I., Saveanu C., Chevalier A., Namane A., Gobin R., Fromont-Racine M., Jacquier A., Mann C. mann@jonas.saclay.cea.fr FRYL genomic DNA library. Mol. Microbiol. (0950-382X) 2002 le_masson-2002-2 Spc24 interacts with Mps2 and is required for chromosome segregation, but is not implicated in spindle pole body duplication. ecobb Escherichia coli (strain B / BL21) pull down pull down anti tag western anti tag western blot anti tag western Le Masson I., Saveanu C., Chevalier A., Namane A., Gobin R., Fromont-Racine M., Jacquier A., Mann C. mann@jonas.saclay.cea.fr FRYL genomic DNA library. Mol. Microbiol. (0950-382X) 2002 le_masson-2002-3 Spc24 interacts with Mps2 and is required for chromosome segregation, but is not implicated in spindle pole body duplication. ecobb Escherichia coli (strain B / BL21) 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 Le Masson I., Saveanu C., Chevalier A., Namane A., Gobin R., Fromont-Racine M., Jacquier A., Mann C. Proteins were purified by nickel affinity chromatography prior to gel filtration on Superdex-200. mann@jonas.saclay.cea.fr FRYL genomic DNA library. Mol. Microbiol. (0950-382X) 2002 le_masson-2002-4 Spc24 interacts with Mps2 and is required for chromosome segregation, but is not implicated in spindle pole body duplication. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Le Masson I., Saveanu C., Chevalier A., Namane A., Gobin R., Fromont-Racine M., Jacquier A., Mann C. mann@jonas.saclay.cea.fr FRYL genomic DNA library. Mol. Microbiol. (0950-382X) 2002 spc24_yeast Kinetochore protein SPC24 SPC24 YM9718.16C YMR117C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSQKDNLLDNPVEFLKEVRESFDIQQDVDAMKRIRHDLDVIKEESEARISKEHSKVSESNKKLNAERINVAKLEGDLEYTNEESNEFGSKDELVKLLKDLDGLERNIVSLRSELDEKMKLYLKDSEIISTPNGSKIKAKVIEPELEEQSAVTPEANENILKLKLYRSLGVILDLENDQVLINRKNDGNIDILPLDNNLSDFYKTKYIWERLGK 8BE4B8619DD4C8FB mps2_yeast Monopolar spindle protein 2 YGL075C MPS2 MMC1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSNGAFDAIFEYAWGQIDKPISGDFIYGKDLPKLIEIIENIFQKAQKSGSYELRLPLFSEINKDLFRTFSNTKTFFKIHKEEFDDIFFNLVNHPLREILENAFIGVDSIPSDFIVSMNLNSPSKFLVENKNKNTEGAGISTPRKKLTESPIKLLSRNNIGKALEVQVEELKRELTAKQSLLQENERQVSELKIRLETYQEKYASIQQRFSDLQKARQVEDNQNSSRTSDPGSPLVTGIDQKAILEEFRRRLQRQTDTISFLKDQIRRERGLNCSNDKVSHSKRKHATTDGDGTFKNFISAVPSNIWVKATIRIIVCFALLAGVLPYIRKYVYAHDTPSQNSRLQLSWWENSGILSKIVWFFEDQTDLETEYRSNANVDDAYSRVFGI bbp1_yeast Spindle pole component BBP1 BBP1 YPL255W P0745 BFR1-binding protein 1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MNQEDNTGGGGIFGLFKWTKDALFGTDISPSMKYKDQEERRDRSRYAQDDTNFSMKFGNDSNRRSTNLSRSNSWSGLDSTLHRKYELLPEYNENGFNSIVNGDHHSKERIRSLRSPAPIVPREPLRNEPTDTFGHRLHTKRRTINELSNSQIPFIPPQEDDPLLSKLFNKDGVNEVRRSPYKLSVKDIPGKFPSPLTKRDEIDNYYVRDEDACHKNREYKKAYFDLFAQMDLNSRDLEDLCEDVREQREQFHRNEQTYKQAYEEMRAELVNELKKSKTLFENYYSLGQKYKSLKKVLDQTISHEAELATSRERLYQEEDLKNFEIQTLKQRLSDLELKYTNLQIEKDMQRDNYESEIHDLLLQLSLRNNERKDTSAGSNIFSTGQ 3A1A827E7664E4E0 af9_yeast Protein AF-9 homolog N1966 YNL107W YAF9 protein protein yeast Saccharomyces cerevisiae Baker's yeast MAPTISKRIKTLSVSRPIIYGNTAKKMGSVKPPNAPAEHTHLWTIFVRGPQNEDISYFIKKVVFKLHDTYPNPVRSIEAPPFELTETGWGEFDINIKVYFVEEANEKVLNFYHRLRLHPYANPVPNSDNGNEQNTTDHNSKDAEVSSVYFDEIVFNEPNEEFFKILMSRPGNLLPSNKTDDCVYSKQLEQEEIDRIEIGIEKVDKEIDELKQKLENLVKQEAINGS 1FE08F72942061D1 spc25_yeast Kinetochore protein SPC25 YER018C SPC25 protein protein yeast Saccharomyces cerevisiae Baker's yeast MASIDAFSDLERRMDGFQKDVAQVLARQQNHARQQLQQFQAEMRQLHNQHQHLIDELQRLATQRTALQQQIHAAQQATNTTREQWRSYHERESELSRRQSTLAAQSRELDSLLQQRGKECVQLRARWAAQSGNDAAEVALYERLLQLRVLPGASDVHDVRFVFGDDSRCWIEVAMHGDHVIGNSHPALDPKSRATLEHVLTVQGDLAAFLVVARDMLLASL 1BED6CCFF330A5D8 ndc80_yeast Kinetochore protein NDC80 NDC80 80 kDa spindle component protein HEC1 Nuclear division cycle protein 80 TID3 Two-hybrid interaction with DMC1 protein 3 YIL144W protein protein yeast Saccharomyces cerevisiae Baker's yeast MQSSTSTDQHVLHHMDPHRFTSQIPTATSSQLRRRNSTNQGLTDMINKSIARNTISGTGIPTGGINKNKRTRSTVAGGTNGTALALNDKSNSRNSVSRLSINQLGSLQQHLSNRDPRPLRDKNFQSAIQEEIYDYLKKNKFDIETNHPISIKFLKQPTQKGFIIIFKWLYLRLDPGYGFTKSIENEIYQILKNLRYPFLESINKSQISAVGGSNWHKFLGMLHWMVRTNIKLDMCLNKVDRSLINQNTQEITILSQPLKTLDEQDQRQERYELMVEKLLIDYFTESYKSFLKLEDNYEPSMQELKLGFEKFVHIINTDIANLQTQNDNLYEKYQEVMKISQKIKTTREKWKALKSDSNKYENYVNAMKQKSQEWPGKLEKMKSECELKEEEIKALQSNISELHKILRKKGISTEQFELQNQEREKLTRELDKINIQSDKLTSSIKSRKLEAEGIFKSLLDTLRQYDSSIQNLTRSRSQLGHNVNDSSLKINISENLLDRDFHEGISYEQLFPKGSGINESIKKSILKLNDEIQERIKTIEKDNITLEKDIKNLKHDINEKTQINEKLELELSEANSKFELSKQENERLLVAQRIEIEKMEKKINDSNLLMKTKISDAEELVTSTELKLEELKVDLNRKRYKLHQQVIHVIDITSKFKINIQSSLENSENELGNVIEELRNLEFETEHNVTN 7DBC492227A80093 mps2_spc24_1 509711 N/A 509716 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 false N/A 509717 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 false physical association physical association false false false Fig. 1 mps2_bbp1_1 509711 N/A 509718 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 false N/A 509717 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 false physical association physical association false false false Fig. 1 mps2_yaf9_1 509711 N/A 509719 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 false N/A 509717 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 false physical association physical association false false false Fig. 1 spc24_spc25_4 509711 N/A 509720 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 false N/A 509716 unspecified role unspecified role bait bait physical association physical association false false false Fig. 1 spc24_ndc80_3 509711 N/A 509721 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 false N/A 509716 unspecified role unspecified role bait bait physical association physical association false false false Fig. 1 spc25_spc24_4 509711 N/A 509720 unspecified role unspecified role bait bait N/A 509716 unspecified role unspecified role prey prey physical association physical association false false false Fig. 3A spc25_ndc80_3 509711 N/A 509720 unspecified role unspecified role bait bait N/A 509721 unspecified role unspecified role prey prey physical association physical association false false false Fig. 3A spc24_ndc80_4 509712 N/A 509720 unspecified role unspecified role prey prey N/A 509721 unspecified role unspecified role prey prey N/A 509716 unspecified role unspecified role bait bait c-terminus 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 association association false false false Fig. 3B ndc80_spc24_3 509712 N/A 509720 unspecified role unspecified role prey prey N/A 509721 unspecified role unspecified role bait bait c-terminus 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 N/A 509716 unspecified role unspecified role prey prey association association false false false Fig. 3B mps2_spc24_2 509713 N/A 509717 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 false n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 509716 unspecified role unspecified role prey prey c-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Fig. 2A mps2_spc24_3 509714 N/A 509716 unspecified role unspecified role prey prey c-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 509717 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 false n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Fig. 2B spc24_mps2_1 509715 N/A 509716 unspecified role unspecified role bait bait c-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 509717 unspecified role unspecified role prey prey n-terminus 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 physical association physical association false false false Fig. 2D