UniProt UniProt http://www.uniprot.org mochida-2004-3 Regulation of checkpoint kinases through dynamic interaction with Crb2. schpm Schizosaccharomyces pombe (fission yeast) Fission yeast pull down pull down western blot western blot Immuno blot Immuno blot Mochida S., Esashi F., Aono N., Tamai K., O'Connell MJ., Yanagida M. yanagida@kozo.biophys.kyoto-u.ac.jp EMBO J. (0261-4189) 2004 mochida-2004-1 Regulation of checkpoint kinases through dynamic interaction with Crb2. in vitro In vitro in gel kinase assay in-gel kinase assay in gel kinase assay predetermined predetermined participant predetermined Mochida S., Esashi F., Aono N., Tamai K., O'Connell MJ., Yanagida M. yanagida@kozo.biophys.kyoto-u.ac.jp EMBO J. (0261-4189) 2004 mochida-2004-2 Regulation of checkpoint kinases through dynamic interaction with Crb2. in vitro In vitro in gel kinase assay in-gel kinase assay in gel kinase assay predetermined predetermined participant predetermined Mochida S., Esashi F., Aono N., Tamai K., O'Connell MJ., Yanagida M. Self-phosphorylation was inferred from apparent shift in molecular weight following western blot. yanagida@kozo.biophys.kyoto-u.ac.jp EMBO J. (0261-4189) 2004 mochida-2004-4 Regulation of checkpoint kinases through dynamic interaction with Crb2. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Mochida S., Esashi F., Aono N., Tamai K., O'Connell MJ., Yanagida M. yanagida@kozo.biophys.kyoto-u.ac.jp EMBO J. (0261-4189) 2004 mochida-2004-5 Regulation of checkpoint kinases through dynamic interaction with Crb2. schpm Schizosaccharomyces pombe (fission yeast) Fission yeast tap tandem affinity purification TAP TAP tap polyclonal western polyclonal antibody western blot polyclonal western Mochida S., Esashi F., Aono N., Tamai K., O'Connell MJ., Yanagida M. yanagida@kozo.biophys.kyoto-u.ac.jp EMBO J. (0261-4189) 2004 rhp9_schpo DNA repair protein crb2 crb2 Checkpoint mediator protein crb2 Cut5-repeat binding protein 2 RAD9 protein homolog rhp9 SPBC342.05 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MEVNDTSLHKGFGLDINSQRVFGAQAAISRNNYSKVNASINPSPPRSNDNSNKEFSYSKDVNNNGAVEELSLTQLFEVPSQAAFAKQSSQDISDDELIQHDSRKVISSPYSPKQTHTVLKRLYDRQSVISDHEKLLTPQNVSNSSQILSPFTSLLPSTLSTLKDTPLSVSQNEKNLETVGEVLVPETVAQHRTKFYDYTLDEMENETESGQVETTPTRLATSLGSPVLYGRVESTPPAFLPETSEKQYKRKFSFTEPSSEKVDNTETKFSKKTKNINDENFPNPFNVISSYETSASPSTVIDQSSQVSSIFVNKRLRKSVNNQAISRSDSLSLDTPKIDSLFTRASIKPLKPSQSPNSRRSFKNRVLAFFKGYPSFYYPATLVAPVHSAVTSSIMYKVQFDDATMSTVNSNQIKRFFLKKGDVVQSTRLGKIKHTVVKTFRSTNEQLSLIAVDALNNDMVILAHGEIEVTVPISTIYVAPVNIRRFQGRDLSFSTLKDMKFEETSFLPSHDSQRNRSSLKERDSSFVKKNLDSESNQLIFDDCVFAFSGPVHEDAYDRSALETVVQDHGGLVLDTGLRPLFNDPFKSKQKKLRHLKPQKRSKSWNQAFVVSDTFSRKVKYLEALAFNIPCVHPQFIKQCLKMNRVVDFSPYLLASGYSHRLDCTLSQRIEPFDTTDSLYDRLLARKGPLFGKKILFIIPEAKSWQKKIENTEQGQKALAHVYHALALGADVEIRPNVAHLECDLILTMDGNIVDETNCPVVDPEWIVECLISQSDIST rad3_schpo Protein kinase rad3 DNA repair protein rad3 rad3 SPBC216.05 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MSQHAKRKAGSLDLSPRGLDDRQAFGQLLKEVLALDKEHELGRSNSLPSMTSELVEVLIEVGLLAFKHDDSKSEFISPKMLKEAHLSLQALMLILKRSPTVLREIKSSVTLLDWILPRTISLFADIRFIKLFDSLKEFHKLIYQLISEKSFLWDLYASFMRYWKYYITNVSSIVLQITNATFPYKMPSPNSQPLQSITPNYPTHREDKFDLLIINIEEACTFFFESAHFFAQCSYLKKSNFPSPPLFTAWTWIKPCFFNFVILLKRISIGDSQLFLHLHSRIVQTLCCFSLNFIYHGLPICEKSKHILMSSINLTLGSLKKTYTVANTAISLFFLSLFVLPKTVAGLFYPFGVSLLSDFKVLEQLEPDSDLKKAIILFKCRYQSSEIDQTTLRAFGEICTGKLENTLFSNSELNLFLLHYLSLDNDLSNILKVDFQNGHNICTFAKWCINNNLDEPSNLKHFREMLDYYSSHNVTISEDDLKNFSLVLCTHVAKVNEKTNSIFRTYEVHGCEVCNSFCLLFDERSLFKIPYHELFCALLKNPDIISSSVKQSLLLDGFFRWSQHCSNFNKESMLSLREFIMKALASTSRCLRVVAAKVLPIFIKGPNNLDIVEFHKESKALIFNTLKILAVENTAILETVILSWISLSRVVEEEELHFVLLEVISSVINSGIFYQGIGLSALQQIASTRHISVWQLLSPYWPTVSVAIVQGMGKKPNIASLFAQLMNISEGDFLIRTQAYTLPFLVLTKNKALIVRIAELSQSDVATLCLTNMHKILASLLTTDHPNLEESVMLLLSLATSDFEKVDLTSLLRSDPISITVELLQLYQNDVPHEKIENALRKVAMIVSQVVNDEDLSNKELLYDFFNNHILGILAEFSNILNDLKGKTSINEKIKTIVGIEKMLSLCGGAVKLGLPQILSNLQSAFQNEHLRFYAIKAWFSLILATKEPEYSSIAGLSLVILPPLFPYLEPQEAELVIQIFDFISSDTHKCLQGLKWAIPTSLDSACFSLKAKEIFCSLQNEDFYSELQSIIKCLTNENEPVCYLGLQKLELFFQAKVDELHDTLNLDISNEVLDQLLRCLLDCCVKYASTNMQISYLAAKNLGELGAIDPSRAKAQHIIKETVVLDNFENGEESLKFILDFMQSQLIPAFLVTTDTKAQGFLAYALQEFLKLGGFKSAVINKKKGLTVVTEHWMSLPDLSKRVLIPFLTSKYHLTPIPKIDIRYPIYKENVTIHTWMQLFSLKLMEYAHSQNAEKIFGICSKVVKDQEVNIPCFLLPFLVLNVILTESELEVNKVIEEFQLVINQPGPDGLNSVGQQRYTSFVDVFFKIVDYLNKWLRMRKKRNWDRRSAIARKENRYMSVEDATSRESSISKVESFLSRFPSKTLGIVSLNCGFHARALFYWEQHIRNATAPYAALESDYRVLQEIYAGIDDPDEIEAVSLNFHDYSFDQQLLLHENSGTWDSALSCYEIIIQKDPENKKAKIGLLNSMLQSGHYESLVLSLDSFIINDNHEYSKMLNLGIEASWRSLSIDSLKKCLSKSNLESFEAKLGSIFYQYLRKDSFAELTERLQPLYVDAATAIANTGAHSAYDCYDILSKLHAINDFSRIAETDGIVSDNLDIVLRRRLSQVAPYGKFKHQILSTHLVGYEKFENTKKTAEIYLEIARISRKNGQFQRAFNAILKAMDLDKPLATIEHAQWWWHQGQHRKAISELNFSLNNNMFDLVDEHEERPKNRKETLGNPLKGKVFLKLTKWLGKAGQLGLKDLETYYHKAVEIYSECENTHYYLGHHRVLMYEEEQKLPVNEQSERFLSGELVTRIINEFGRSLYYGTNHIYESMPKLLTLWLDFGAEELRLSKDDGEKYFREHIISSRKKSLELMNSNVCRLSMKIPQYFFLVALSQMISRVCHPNNKVYKILEHIIANVVASYPGETLWQLMATIKSTSQKRSLRGKSILNVLHSRKLSMSSKVDIKALSQSAILITEKLINLCNTRINSKSVKMSLKDHFRLSFDDPVDLVIPAKSFLDITLPAKDANRASHYPFPKTQPTLLKFEDEVDIMNSLQKPRKVYVRGTDGNLYPFLCKPKDDLRKDARLMEFNNLICKILRKDQEANRRNLCIRTYVVIPLNEECGFIEWVNHTRPFREILLKSYRQKNIPISYQEIKVDLDFALRSPNPGDIFEKKILPKFPPVFYEWFVESFPEPNNWVTSRQNYCRTLAVMSIVGYVLGLGDRHGENILFDEFTGEAIHVDFNCLFDKGLTFEKPEKVPFRLTHNMVDAMGPTGYEGGFRKASEITMRLLRSNQDTLMSVLESFLHDPLVEWNRKKSSSKYPNNEANEVLDIIRKKFQGFMPGETIPLSIEGQIQELIKSAVNPKNLVEMYIGWAAYF chk1_schpo Serine/threonine-protein kinase chk1 Checkpoint kinase 1 chk1 rad27 SPCC1259.13 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MAQKLDNFPYHIGREIGTGAFASVRLCYDDNAKIYAVKFVNKKHATSCMNAGVWARRMASEIQLHKLCNGHKNIIHFYNTAENPQWRWVVLEFAQGGDLFDKIEPDVGIDEDVAQFYFAQLMEGISFMHSKGVAHRDLKPENILLDYNGNLKISDFGFASLFSYKGKSRLLNSPVGSPPYAAPEITQQYDGSKVDVWSCGIILFALLLGNTPWDEAISNTGDYLLYKKQCERPSYHPWNLLSPGAYSIITGMLRSDPFKRYSVKHVVQHPWLTSSTPFRTKNGNCADPVALASRLMLKLRIDLDKPRLASSRASQNDSGFSMTQPAFKKNDQKELDRVEVYGALSQPVQLNKNIDVTEILEKDPSLSQFCENEGFIKRLAKKAKNFYEICPPERLTRFYSRASRETIIDHLYDSLRLLAISVTMKYVRNQTILYVNLHDKRKCLLQGVIELTNLGHNLELINFIKRNGDPLEWRKFFKNVVSSIGKPIVLTDVSQN mpip_schpo M-phase inducer phosphatase cdc25 SPAC24H6.05 Mitosis initiation protein P80 protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MDSPLSSLSFTNTLSGKRNVLRPAARELKLMSDRNANQELDFFFPKSKHIASTLVDPFGKTCSTASPASSLAADMSMNMHIDESPALPTPRRTLFRSLSCTVETPLANKTIVSPLPESPSNDALTESYFFRQPASKYSITQDSPRVSSTIAYSFKPKASIALNTTKSEATRSSLSSSSFDSYLRPNVSRSRSSGNAPPFLRSRSSSSYSINKKKGTSGGQATRHLTYALSRTCSQSSNTTSLLESCLTDDTDDFELMSDHEDTFTMGKVADLPESSVELVEDAASIQRPNSDFGACNDNSLDDLFQASPIKPIDMLPKINKDIAFPSLKVRSPSPMAFAMQEDAEYDEQDTPVLRRTQSMFLNSTRLGLFKSQDLVCVTPKQSTKESERFISSHVEDLSLPCFAVKEDSLKRITQETLLGLLDGKFKDIFDKCIIIDCRFEYEYLGGHISTAVNLNTKQAIVDAFLSKPLTHRVALVFHCEHSAHRAPHLALHFRNTDRRMNSHRYPFLYYPEVYILHGGYKSFYENHKNRCDPINYVPMNDASHVMTCTKAMNNFKRNATFMRTKSYTFGQSVLASPDVNDSPTAMHSLSTLRRF 4ebp1_human Eukaryotic translation initiation factor 4E-binding protein 1 Phosphorylated heat- and acid-stable protein regulated by insulin 1 EIF4EBP1 protein protein human Homo sapiens Human MSGGSSCSQTPSRAIPATRRVVLGDGVQLPPGDYSTTPGGTLFSTTPGGTRIIYDRKFLMECRNSPVTKTPPRDLPTIPGVTSPSSDEPPMEASQSHLRNSPEDKRAGGEESQFEMDI rad4_schpo S-M checkpoint control protein rad4 Protein cut5 P74 rad4 cut5 SPAC23C4.18c protein protein schpo Schizosaccharomyces pombe (strain ATCC 38366 / 972) Fission yeast MGSSKPLKGFVICCTSIDLKQRTEISTKATKLGAAYRSDFTKDVTHLIAGDFDTPKYKFAAKSRPDIKIMSSEWIPVLYESWVQGEDLDDGLLVDKHFLPTLFKCRVCLTNIGQPERSRIENYVLKHGGTFCPDLTRDVTHLIAGTSSGRKYEYALKWKINVVCVEWLWQSIQRNAVLEPQYFQLDMPAEKIGLGAYVRLDPNTTEAKSYSENQKISKNKEKSGQSLAALAEEADLEPVIMKRGKKRDRSILWEELNNGKFEFSSRSEENSVLLDDFTPETVQPLEENELDTELNIENEAKLFKNLTFYLYEFPNTKVSRLHKCLSDNGGQISEFLSSTIDFVVIPHYFPVDELPIFSFPTVNEWWIERCLYYKKIFGIDEHALAKPFFRPSLVPYFNGLSIHLTGFKGEELSHLKKALTILGAVVHEFLGVQRSILLVNTNEPFSMKTRFKIQHATEWNVRVVGVAWLWNIIQSGKFIDQVSPWAIDKKENQEIKKFTNQNNMVFPTSDRDTRLQNSLAQQPIGHSTPHNSPSLLSVKKRQNNHIRSNTLIQLNSNSKDSTIFPRRSVTVPGDKIDTVWKSSVTKPETPTSPQEHVSYIDPDAQREKHKLYAQLTSNVDAIPPANDLQNQENGLLLITESHRKLRRR rad3-rhp9-1 567123 N/A 567128 unspecified role unspecified role prey prey N/A 567129 unspecified role unspecified role bait bait n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst 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. 3A rad3-rhp9-2 567123 N/A 567128 unspecified role unspecified role prey prey n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 567129 unspecified role unspecified role bait bait n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false 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. 3B, 3C, 4A, 4B This interaction is transiently reduced following UV irradiation. rad3-chk1-1 567123 N/A 567130 unspecified role unspecified role prey prey n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 567129 unspecified role unspecified role bait bait n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst 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. 4C, 4D This interaction is transiently reduced following UV irradiation. chk1-rhp9-1 567123 N/A 567130 unspecified role unspecified role bait bait n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 567128 unspecified role unspecified role prey prey physical association physical association false false false Fig. 5A, 5B, 5C This interaction is transiently enhanced following UV irradiation. chk1-cdc25-1 567124 N/A 567131 enzyme target enzyme target substrate substrate unspecified role unspecified role region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) N/A 567128 unspecified role unspecified role ancillary ancillary region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 567130 enzyme enzyme unspecified role unspecified role 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 P34208:p.Asp155Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast phosphorylation phosphorylation reaction phosphorylation false false false Fig. 1A, 1B, 1C This interaction is stimulated by UV irradiation or loss of function mutations in swi7, cdc6 or cdc20. rad3-eif4ebp1-1 567124 N/A 567128 inhibitor inhibitor unspecified role unspecified role region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P87074:p.Ser666Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 567132 enzyme target enzyme target substrate substrate unspecified role unspecified role N/A 567129 enzyme enzyme unspecified role unspecified role n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q02099:p.Asp2249Glu mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast phosphorylation phosphorylation reaction phosphorylation false false false Fig. 6 chk1-chk1-1 567125 N/A 567128 unspecified role unspecified role ancillary ancillary region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 567130 enzyme enzyme unspecified role unspecified role P34208:p.Asp155Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false 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 schpm Schizosaccharomyces pombe (fission yeast) Fission yeast N/A 567130 enzyme target enzyme target substrate substrate unspecified role unspecified role 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 schpm Schizosaccharomyces pombe (fission yeast) Fission yeast phosphorylation phosphorylation reaction phosphorylation false false false Fig. 1A, 1B, 1C This interaction is stimulated by UV irradiation or loss of function mutations in swi7, cdc6 or cdc20. rad3-rhp9-3 567126 N/A 567129 unspecified role unspecified role bait bait n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst 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 N/A 567128 unspecified role unspecified role prey prey n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false schpm Schizosaccharomyces pombe (fission yeast) Fission yeast physical association physical association false false false Fig. 4F This interaction is reduced following prior UV irradiation of cells expressing rhp9/crb2. rhp9-cut5-1 567127 N/A 567133 unspecified role unspecified role prey prey N/A 567128 unspecified role unspecified role bait bait n-terminus tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association false false false Fig. 5D This interaction is enhanced following UV irradiation.