MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Dephosphorylation of Cdc20 is required for its C-box-dependent activation of the APC/C. Dephosphorylation of Cdc20 is required for its C-box-dependent activation of the APC/C. EMBO J. (0261-4189) 2012 imex curation Labit H., Fujimitsu K., Bayin NS., Takaki T., Gannon J., Yamano H. Only protein-protein interactions in vitro In vitro pull down pull down experimental particp experimental participant identification experimental particp Labit H., Fujimitsu K., Bayin NS., Takaki T., Gannon J., Yamano H. EMBO J. (0261-4189) 2012 Only protein-protein interactions imex curation Dephosphorylation of Cdc20 is required for its C-box-dependent activation of the APC/C. Dephosphorylation of Cdc20 is required for its C-box-dependent activation of the APC/C. EMBO J. (0261-4189) 2012 imex curation Labit H., Fujimitsu K., Bayin NS., Takaki T., Gannon J., Yamano H. Only protein-protein interactions xenla- Xenopus laevis egg egg anti tag coip anti tag coimmunoprecipitation anti tag coip experimental particp experimental participant identification experimental particp Labit H., Fujimitsu K., Bayin NS., Takaki T., Gannon J., Yamano H. EMBO J. (0261-4189) 2012 Only protein-protein interactions imex curation Dephosphorylation of Cdc20 is required for its C-box-dependent activation of the APC/C. Dephosphorylation of Cdc20 is required for its C-box-dependent activation of the APC/C. EMBO J. (0261-4189) 2012 imex curation Labit H., Fujimitsu K., Bayin NS., Takaki T., Gannon J., Yamano H. Only protein-protein interactions xenla- Xenopus laevis egg egg anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Labit H., Fujimitsu K., Bayin NS., Takaki T., Gannon J., Yamano H. EMBO J. (0261-4189) 2012 Only protein-protein interactions imex curation q8avg7_xenla cdc20.S cdc20 XELAEV_18025163mg protein protein xenla Xenopus laevis (African clawed frog) African clawed frog MAQFAFETDINSILKLDTPITNAPLARWQRKAKEGNCSLNTSANTSTMSPMKTSNRSHSSSKTPSKTPGKSGQKMQGTPSRAGGDRFIPNRSAMQMDVASFLLSKENEPTDTSPTKKEQQKAWAMNLNGFDMEEAKILRLGGRPQNAPEGYQNNLKVLYSQKNTPGSSKKTGRYIPSMPDRVLDAPDIRNDYYLNLIDWSSQNALAVALNDSVYLWNYATGDIILLLQMENSEEYISSVSWIKEGHFLAVGTSNSEVQLWDVQQQKRLRNMISHSSRVGALSWNNHILSSGSRTGHIHHHDVRVAQHHVSTLTGHTQEVCGLKWSPDGRYLASGANDNLVNVWPCVQGDSGEFSPVQTFTQHQGAVKAVAWCPWQSNVLATGGGTSDRHIRIWNVCSGTCLNSVDTHSQVCSILWSANYKELISGHGFAQNQLVLWKYPTMTRVSELKGHTARVLNLAMSPDGCTVASAAADETLRLWKCFEVDPVTKKEKEKSRSSKSIIHQSIR mint DC2D3483521045D0 q7t0n9_xenla cdc23.L cdc23 protein protein xenla Xenopus laevis (African clawed frog) African clawed frog MAATIGMFSDLKEIKRQLLSVTWLCRERGLMNSVKWASELAFSLESVPLNELPSTPALTEEDAQDLDAYMLAKSYFDLKEYDRAAYFLRGCKSQKAYFMYMYSRYLSGEKKKDDETVDSLGPLEKGQVKNEALREVRVELSKKHKARELDGFGLYLYGVVLRKLDLAKEALDVFVEATHVLPLHWGTWLELCNLITDKEMLKFLSLPDSWIKEFFLAHIYTELQLIEEALQKYQSLIDAGFSKSTYIISQIAVAYHNIRDIDKALSIFNELRKQDPYRIENMDTFSNLLYVRGLKPELSYLAHNLCDIDKYRVETCCVIGNYYSLRSQHEKAALYFQRALKLNPRYLGAWTLMGHEYMEMKNTSAAIQAYRHAIEVNKRDYRAWYGLGQTYEILKMPFYCLYYYRRAHQLRPNDSRMLVALGECYEKLNQLVEAKKCYWRAYAVGDVEKMALVKLAKLHEQLNESEQAAQCYIKYIQDIYSCGEIVEHQEVSTAFRYLAQYYFKCKLWDEASACAQKCFNFNDTREEGKALLRQILQLRNQSESPSAEPATAPFFLPQSLSANNTPTRRVSPLNLSSVTP AF133D82C9C52118 q0p4v8_xentr cdc27 MGC145551 protein protein xentr Xenopus tropicalis (Western clawed frog) Silurana tropicalis MTVLQEPVQAAIWQALNHYAYRDAVFLAERLYAEVHSEEALFLLATCYYRSGKAYKAYRLLKAHSCATPQCKYLLAKCCVDLSKLAEGEQILSGGVLNKQKSQEEIVTEFGDSACFTLSLLGHVYCKTDRLAKGAECYQKSLSLNPFLWSPFESLCEIGEKPDPEQTFKLTSLQNFSNCLPNTCTMTVNTHSVVQHRQPDTVLMETPQDTIELNRLNLESSNMKYPLNSDSSISYIDPGVISPDSLPLGTGTSILSKQAQNKPKTGRSLLGGPAALSPLTPSFGILPLETPSPGDGSYLQTYSSCVLDVPSSGAPTKKSVARISQAGTKSVFSQSGNSRDVTPVLLVQTQGSGPQTSTTPQVLSPTIAAPPNALPRRSSRLFTSDSSTTKENSKKLKMKFPPKMPNRKTKSKTSKGGVTQANMNDSLEISKLDSSIISEGKISSVAPQIQALTIQKAAAEGLMSLLRDMGKGYVALCSYNCKEAINILSHLPSHHYNTGWVLCQIGRAYFELAEYLQAERVFSEVRRIESYRVEGMEIYSTTLWHLQKDVTLSVLSKDLTDMDKNSPEAWCAAGNCFSLQREHDIAIKFFQRAIQVDPGYAYAFTLLGHEFVLTEELDKALACFRNAIRVNPRHYNAWYGLGMIYYKQEKFSLAEMHFQKALDINPQSSVLLCHIGVVQHALKKSDKALDTLNKAISIDPKNPLCKFHRASILFANEKYKLALQELEELKQIVPKESLVYFLIGKVYKKLGQTHLALMNFSWAMDLDPKGANNQIKEAIDKRYLPDDEEAVTQEEQISECYPYESVATDESQESSMTDADDTQLHAAESDEF mint C75A42DC0FFA72CB q6dfi7_xenla cdc16.L cdc16 cdc16-prov XELAEV_18012976mg protein protein xenla Xenopus laevis (African clawed frog) African clawed frog MNLDRLRKRVRQYIEQQQYQSALFWADKVASLSHEEPQDIYWLAQCLYLTAQYHRASHALRSRKLDKVHEACRYLAARCHYAAKEYQQALDILDMEEPINKKLFEKSLKEDLKEPSGEWQMSQSSIKSSICLLRGKIYDALDNRTLATYSYKEALKFDVYCFEAFDLLTAHHMLTAQEEKELLDSLPLSKKCSEDQELLRFLYENKLKKYNKPSETLIPESADGLQENLDVVVSLAERHYYNCDFKMCYKLTSVVMEKDPFHANCLPVHIGTLVELNKANELFYLSHKLVDLYPNNPVSWFAVGCYYLMVGHKNEHARRYLSKATALERSYGPAWIAYGHSFAVESEHDQAMAAYFTAAQLMKGCHLPMLYIGLEYGLTNNSKLAERFFSQALSIAPEDPFVMHEVGVVAFQNGDWKTAEKWFLDALEKIRVIGDEVTVDKWEPLLNNLGHVLRKLKKYDEALEYHRQALVLIPQNASTYSAIGYIHSLMGNFESAIDYFHTALGLRRDDTFSVTMLGHCIEMYIGDSDAYIGTEIKDKLRCYDFDVHTMKTLKNIISPPWEEREFDFERQSLDESGIMALETSNLLPKSSDTRSPIEEALEIEMNESDMMLETSMSDHST DIP protein Q6DFI7 original sequence version: 38 8BEC989BCF0C78DF cdc20-cdc16 1 cdc20 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined undefined mutation mutation increasing interaction mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:36:14 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation increasing' Thr64Ala undefined mutation mutation increasing interaction mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:36:13 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation increasing' Ser50Ala undefined mutation mutation increasing interaction mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:36:12 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation increasing' Thr79Ala undefined mutation mutation increasing interaction mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:36:34 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation increasing' Thr68Ala undefined mutation mutation increasing interaction mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:36:33 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation increasing' Ser114Ala Apc3 6 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level Apc6 7 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level association association mint Finally, we investigated the relationship between the C-box (Cb)- and dephosphorylation-dependent association of Cdc20 with the APC/C. We prepared GST-fused N159 with DCb, 5A, 5A-DCb and WT (Figure 6A), incubated these with CSF extracts and monitored the interaction with the APC/C using Apc3 and Apc6 antibodies. As expected, WT, but not DCb, bound the APC/C and pulled down the APC/C complex (Figure 6B). In contrast, the same fragment with 5A showed a much higher affinity and could pull down 410 times more APC/C than WT (Figure 6B). Intriguingly, even 5A-DCb was able to bind more APC/C than WT, although it lacks the C-box. f2a f6b cdc20-cdc16-1 1 cdc20 4 weight by staining molecular weight estimation by staining weight by staining experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Apc6 7 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f6c mint cdc20-cdc23 1 cdc20 4 weight by staining molecular weight estimation by staining weight by staining experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Apc8 5 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f6c mint cdc27-cdc20-3 3 cdc20 4 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Apc3 6 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f2c mint cdc23-cdc20 2 cdc20 4 anti tag western anti tag western blot anti tag western experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Apc8 5 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q7T0N9:p.Asn321Ala mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain N A Tue Jul 12 10:51:40 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 14:39:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn321ala' Tue Jun 30 20:13:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn321Ala' physical association physical association mint Since both 5A and 5E bind the Apc8 subunit, we investigated how the N-terminal fragments interact with the Apc8 subunit. Both GST-N159 WT and 5A fragments bound Apc8 equally; however, the interaction was abolished if the C-box was mutated (Figure 6D). To confirm the interaction between the C-box and Apc8, we introduced a point mutation in the TPR domain of Apc8 (N321A) that had previously been shown to reduce binding to the Fizzy/Cdc20 family (Matyskiela and Morgan, 2009; Izawa and Pines, 2011). Figure 6E shows that the point mutation of the TPR in Apc8 eliminated the C-box-dependent binding. f6d f6e