UniProt UniProt http://www.uniprot.org Complex formation between Mad1p, Bub1p and Bub3p is crucial for spindle checkpoint function. Complex formation between Mad1p, Bub1p and Bub3p is crucial for spindle checkpoint function. Curr. Biol. (0960-9822) 2000 Brady DM., Hardwick KG. For anti-MAD1 precipitation, wild-type cells were arrested in G1 phase with alpha factor, washed and then released into fresh media in the presence or absence of nocodazole for time course analyses. For anti-BUB1 precipitation checkpoint mutant strains were treated with nocodazole for 3 h before extracts were made. hardwick@holyroad.ed.ac.uk 13-Nov-2007: Contacted by JYOTI. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti bait coip anti bait coimmunoprecipitation anti bait coip antibody detection identification by antibody antibody detection Brady DM., Hardwick KG. For anti-MAD1 precipitation, wild-type cells were arrested in G1 phase with alpha factor, washed and then released into fresh media in the presence or absence of nocodazole for time course analyses. For anti-BUB1 precipitation checkpoint mutant strains were treated with nocodazole for 3 h before extracts were made. hardwick@holyroad.ed.ac.uk Accepted 2007-NOV-22 by JYOTI. 13-Nov-2007: Contacted by JYOTI. Curr. Biol. (0960-9822) 2000 Complex formation between Mad1p, Bub1p and Bub3p is crucial for spindle checkpoint function. Complex formation between Mad1p, Bub1p and Bub3p is crucial for spindle checkpoint function. Curr. Biol. (0960-9822) 2000 Brady DM., Hardwick KG. For anti-MAD1 precipitation, wild-type cells were arrested in G1 phase with alpha factor, washed and then released into fresh media in the presence or absence of nocodazole for time course analyses. For anti-BUB1 precipitation checkpoint mutant strains were treated with nocodazole for 3 h before extracts were made. hardwick@holyroad.ed.ac.uk 13-Nov-2007: Contacted by JYOTI. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Brady DM., Hardwick KG. For anti-MAD1 precipitation, wild-type cells were arrested in G1 phase with alpha factor, washed and then released into fresh media in the presence or absence of nocodazole for time course analyses. For anti-BUB1 precipitation checkpoint mutant strains were treated with nocodazole for 3 h before extracts were made. hardwick@holyroad.ed.ac.uk Accepted 2007-NOV-22 by JYOTI. 13-Nov-2007: Contacted by JYOTI. Curr. Biol. (0960-9822) 2000 Complex formation between Mad1p, Bub1p and Bub3p is crucial for spindle checkpoint function. Complex formation between Mad1p, Bub1p and Bub3p is crucial for spindle checkpoint function. Curr. Biol. (0960-9822) 2000 Brady DM., Hardwick KG. For anti-MAD1 precipitation, wild-type cells were arrested in G1 phase with alpha factor, washed and then released into fresh media in the presence or absence of nocodazole for time course analyses. For anti-BUB1 precipitation checkpoint mutant strains were treated with nocodazole for 3 h before extracts were made. hardwick@holyroad.ed.ac.uk 13-Nov-2007: Contacted by JYOTI. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Brady DM., Hardwick KG. For anti-MAD1 precipitation, wild-type cells were arrested in G1 phase with alpha factor, washed and then released into fresh media in the presence or absence of nocodazole for time course analyses. For anti-BUB1 precipitation checkpoint mutant strains were treated with nocodazole for 3 h before extracts were made. hardwick@holyroad.ed.ac.uk Accepted 2007-NOV-22 by JYOTI. 13-Nov-2007: Contacted by JYOTI. Curr. Biol. (0960-9822) 2000 Complex formation between Mad1p, Bub1p and Bub3p is crucial for spindle checkpoint function. Complex formation between Mad1p, Bub1p and Bub3p is crucial for spindle checkpoint function. Curr. Biol. (0960-9822) 2000 Brady DM., Hardwick KG. For anti-MAD1 precipitation, wild-type cells were arrested in G1 phase with alpha factor, washed and then released into fresh media in the presence or absence of nocodazole for time course analyses. For anti-BUB1 precipitation checkpoint mutant strains were treated with nocodazole for 3 h before extracts were made. hardwick@holyroad.ed.ac.uk 13-Nov-2007: Contacted by JYOTI. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti bait coip anti bait coimmunoprecipitation anti bait coip antibody detection identification by antibody antibody detection Brady DM., Hardwick KG. For anti-MAD1 precipitation, wild-type cells were arrested in G1 phase with alpha factor, washed and then released into fresh media in the presence or absence of nocodazole for time course analyses. For anti-BUB1 precipitation checkpoint mutant strains were treated with nocodazole for 3 h before extracts were made. hardwick@holyroad.ed.ac.uk Accepted 2007-NOV-22 by JYOTI. Curr. Biol. (0960-9822) 2000 Complex formation between Mad1p, Bub1p and Bub3p is crucial for spindle checkpoint function. Complex formation between Mad1p, Bub1p and Bub3p is crucial for spindle checkpoint function. Curr. Biol. (0960-9822) 2000 Brady DM., Hardwick KG. For anti-MAD1 precipitation, wild-type cells were arrested in G1 phase with alpha factor, washed and then released into fresh media in the presence or absence of nocodazole for time course analyses. For anti-BUB1 precipitation checkpoint mutant strains were treated with nocodazole for 3 h before extracts were made. hardwick@holyroad.ed.ac.uk 13-Nov-2007: Contacted by JYOTI. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip antibody detection identification by antibody antibody detection Brady DM., Hardwick KG. For anti-MAD1 precipitation, wild-type cells were arrested in G1 phase with alpha factor, washed and then released into fresh media in the presence or absence of nocodazole for time course analyses. For anti-BUB1 precipitation checkpoint mutant strains were treated with nocodazole for 3 h before extracts were made. hardwick@holyroad.ed.ac.uk Accepted 2007-NOV-22 by JYOTI. 13-Nov-2007: Contacted by JYOTI. Curr. Biol. (0960-9822) 2000 bub3_yeast Cell cycle arrest protein BUB3 OR26.16 YOR026W BUB3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MQIVQIEQAPKDYISDIKIIPSKSLLLITSWDGSLTVYKFDIQAKNVDLLQSLRYKHPLLCCNFIDNTDLQIYVGTVQGEILKVDLIGSPSFQALTNNEANLGICRICKYGDDKLIAASWDGLIEVIDPRNYGDGVIAVKNLNSNNTKVKNKIFTMDTNSSRLIVGMNNSQVQWFRLPLCEDDNGTIEESGLKYQIRDVALLPKEQEGYACSSIDGRVAVEFFDDQGDDYNSSKRFAFRCHRLNLKDTNLAYPVNSIEFSPRHKFLYTAGSDGIISCWNLQTRKKIKNFAKFNEDSVVKIACSDNILCLATSDDTFKTNAAIDQTIELNASSIYIIFDYEN 0BDFB8697935BCEB 0BDFB8697935BCEB mad1_yeast Spindle assembly checkpoint component MAD1 MAD1 Mitotic arrest deficient protein 1 YGL086W protein protein yeast Saccharomyces cerevisiae Baker's yeast MDVRAALQCFFSALSGRFTGKKLGLEIYSIQYKMSNSGGSSPFLESPGGSPDVGSTNGQSNRQIQALQFKLNTLQNEYEIEKLQLQKQTNILEKKYKATIDELEKALNDTKYLYESNDKLEQELKSLKERSANSMNDKDKCIEELRTTLQNKDLEMETLRQQYDSKLSKVTNQCDHFKLEAESSHSLLMKYEKEIKRQSVDIKDLQHQVMEKDDELSSVKASKMINSHPNYSTEEFNELTEMNKMIQDQVQYTKELELANMQQANELKKLKQSQDTSTFWKLENEKLQNKLSQLHVLESQYENLQLENIDLKSKLTKWEIYNDSDDDDDNNVNNNDNNNNNKNDNNNDNNNDTSNNNNINNNNRTKNNIRNNPEEIIRDWKLTKKECLILTDMNDKLRLDNNNLKLLNDEMALERNQILDLNKNYENNIVNLKRLNHELEQQKSLSFEECRLLREQLDGLYSAQNNALLEVENSETHASNKNVNEDMNNLIDTYKNKTEDLTNELKKLNDQLLSNSNDVETQRKKRKLTSDQIGLNYSQRLNELQLENVSVSRELSKAQTTIQLLQEKLEKLTKLKEKKIRILQLRDGPFIKDQFIKKNKLLLLEKENADLLNELKKNNPAVETVPISVYDSLNFELKQFEQEVFKSNKRFSRLKQVFNNKSLEFIDVVNSLLGFKLEFQQDSRVKIFSCFKPEKYLIADLNENTLKSNLDADIEGWDDLMNLWVEDRGQLPCFLATITLRLWEQRQAK 312DDE1BE241610D mad2_yeast Mitotic spindle checkpoint component MAD2 MAD2 J1256 YJL030W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSQSISLKGSTRTVTEFFEYSINSILYQRGVYPAEDFVTVKKYDLTLLKTHDDELKDYIRKILLQVHRWLLGGKCNQLVLCIVDKDEGEVVERWSFNVQHISGNSNGQDDVVDLNTTQSQIRALIRQITSSVTFLPELTKEGGYTFTVLAYTDADAKVPLEWADSNSKEIPDGEVVQFKTFSTNDHKVGAQVSYKY EFE59916C5720644 bub1_yeast Checkpoint serine/threonine-protein kinase BUB1 YGR188C G7542 BUB1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MNLDLGSTVRGYESDKDTFPQSKGVSSSQKEQHSQLNQTKIAYEQRLLNDLEDMDDPLDLFLDYMIWISTSYIEVDSESGQEVLRSTMERCLIYIQDMETYRNDPRFLKIWIWYINLFLSNNFHESENTFKYMFNKGIGTKLSLFYEEFSKLLENAQFFLEAKVLLELGAENNCRPYNRLLRSLSNYEDRLREMNIVENQNSVPDSRERLKGRLIYRTAPFFIRKFLTSSLMTDDKENRANLNSNVGVGKSAPNVYQDSIVVADFKSETERLNLNSSKQPSNQRLKNGNKKTSIYADQKQSNNPVYKLINTPGRKPERIVFNFNLIYPENDEEFNTEEILAMIKGLYKVQRRGKKHTEDYTSDKNRKKRKLDVLVERRQDLPSSQPPVVPKSTRIEVFKDDDNPSQSTHHKNTQVQVQTTTSILPLKPVVDGNLAHETPVKPSLTSNASRSPTVTAFSKDAINEVFSMFNQHYSTPGALLDGDDTTTSKFNVFENFTQEFTAKNIEDLTEVKDPKQETVSQQTTSTNETNDRYERLSNSSTRPEKADYMTPIKETTETDVVPIIQTPKEQIRTEDKKSGDNTETQTQLTSTTIQSSPFLTQPEPQAEKLLQTAEHSEKSKEHYPTIIPPFTKIKNQPPVIIENPLSNNLRAKFLSEISPPLFQYNTFYNYNQELKMSSLLKKIHRVSRNENKNPIVDFKKTGDLYCIRGELGEGGYATVYLAESSQGHLRALKVEKPASVWEYYIMSQVEFRLRKSTILKSIINASALHLFLDESYLVLNYASQGTVLDLINLQREKAIDGNGIMDEYLCMFITVELMKVLEKIHEVGIIHGDLKPDNCMIRLEKPGEPLGAHYMRNGEDGWENKGIYLIDFGRSFDMTLLPPGTKFKSNWKADQQDCWEMRAGKPWSYEADYYGLAGVIHSMLFGKFIETIQLQNGRCKLKNPFKRYWKKEIWGVIFDLLLNSGQASNQALPMTEKIVEIRNLIESHLEQHAENHLRNVILSIEEELSHFQYKGKPSRRF 6D76FC980775D3F9 6D76FC980775D3F9 mad1-bub1-1 4 7 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association 1a BUB1 was detected from 40 min to 60 min of the unperturbed cell cycle, and at significantly higher levels in the nocodazole-treated cells from 40 min onwards. Apparently a MAD1-BUB1-BUB3 complex was formed 40 minutes after release from an alpa-factor arrest and was then maintained. mad1-bub3-1 2 6 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role bait bait physical association physical association 1a BUB3 was detected from 40 min to 60 min of the unperturbed cell cycle, and at significantly higher levels in the nocodazole-treated cells from 40 min onwards. Apparently a MAD1-BUB1-BUB3 complex was formed 40 minutes after release from an alpa-factor arrest and was then maintained. bub3-bub1-4 5 6 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role prey prey physical association physical association BUB1 was detected in G1, mitosis and metaphase precipitates. 1b bub3-bub1-5 5 6 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey physical association physical association MAD1 was detected in mitosis precipitates (with activated spindle checkpoint). 1b bub1-mad1-1 4 7 unspecified role unspecified role prey prey P40957:p.[Arg653del;Lys655del] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain R . certain certain sequence position certain certain certain sequence position certain K . Wed May 02 19:32:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg653del,lys655del' Tue Jun 30 16:30:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg653del;Lys655del]' 9 unspecified role unspecified role bait bait physical association physical association 3a Interaction requires MAD2 and BUB3. MAD1 mutation disrupting was taken from fig 4c with anti-MAD1 IP. bub1-bub3-2 4 6 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association 3a Interaction does not require MAD1, MAD2, MAD3, and BUB2. mad1-bub1-2 1 9 unspecified role unspecified role prey prey P41695:p.Lys733Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K R Wed May 02 14:49:57 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys733arg' Tue Jun 30 17:04:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys733Arg' 7 unspecified role unspecified role bait bait physical association physical association Interaction is dependent on MPS1. 3b mad1-bub1-5 3 7 unspecified role unspecified role bait bait region binding region binding-associated region binding region mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain 9 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey association association Detected in medium with nocodozole. A MAD1 653-RLK-AAA mutant did not interact with BUB1 and BUB3 but did not change the interaction with MAD2. 4c