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/ Sgt1p and Skp1p modulate the assembly and turnover of CBF3 complexes required for proper kinetochore function. Sgt1p and Skp1p modulate the assembly and turnover of CBF3 complexes required for proper kinetochore function. Mol. Biol. Cell (1059-1524) 2004 imex curation Rodrigo-Brenni MC., Thomas S., Bouck DC., Kaplan KB. kbkaplan@ucdavis.edu 26-Sep-2011: Contacted by JYOTI. jyoti Only protein-protein interactions trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Rodrigo-Brenni MC., Thomas S., Bouck DC., Kaplan KB. kbkaplan@ucdavis.edu Accepted 2011-JUN-30 by JYOTI 26-Sep-2011: Contacted by JYOTI. Mol. Biol. Cell (1059-1524) 2004 jyoti Only protein-protein interactions imex curation Sgt1p and Skp1p modulate the assembly and turnover of CBF3 complexes required for proper kinetochore function. Sgt1p and Skp1p modulate the assembly and turnover of CBF3 complexes required for proper kinetochore function. Mol. Biol. Cell (1059-1524) 2004 imex curation Rodrigo-Brenni MC., Thomas S., Bouck DC., Kaplan KB. kbkaplan@ucdavis.edu 26-Sep-2011: Contacted by JYOTI. jyoti Only protein-protein interactions trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. pull down pull down western blot western blot Immuno blot Immuno blot Rodrigo-Brenni MC., Thomas S., Bouck DC., Kaplan KB. kbkaplan@ucdavis.edu Accepted 2011-JUN-30 by JYOTI 26-Sep-2011: Contacted by JYOTI. Mol. Biol. Cell (1059-1524) 2004 jyoti Only protein-protein interactions imex curation Sgt1p and Skp1p modulate the assembly and turnover of CBF3 complexes required for proper kinetochore function. Sgt1p and Skp1p modulate the assembly and turnover of CBF3 complexes required for proper kinetochore function. Mol. Biol. Cell (1059-1524) 2004 imex curation Rodrigo-Brenni MC., Thomas S., Bouck DC., Kaplan KB. kbkaplan@ucdavis.edu 26-Sep-2011: Contacted by JYOTI. jyoti Only protein-protein interactions 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 Rodrigo-Brenni MC., Thomas S., Bouck DC., Kaplan KB. kbkaplan@ucdavis.edu Accepted 2011-JUN-30 by JYOTI 26-Sep-2011: Contacted by JYOTI. Mol. Biol. Cell (1059-1524) 2004 jyoti Only protein-protein interactions imex curation Sgt1p and Skp1p modulate the assembly and turnover of CBF3 complexes required for proper kinetochore function. Sgt1p and Skp1p modulate the assembly and turnover of CBF3 complexes required for proper kinetochore function. Mol. Biol. Cell (1059-1524) 2004 imex curation Rodrigo-Brenni MC., Thomas S., Bouck DC., Kaplan KB. kbkaplan@ucdavis.edu 26-Sep-2011: Contacted by JYOTI. 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 Rodrigo-Brenni MC., Thomas S., Bouck DC., Kaplan KB. kbkaplan@ucdavis.edu Accepted 2011-JUN-30 by JYOTI 26-Sep-2011: Contacted by JYOTI. Mol. Biol. Cell (1059-1524) 2004 jyoti Only protein-protein interactions imex curation cbf3a_yeast Centromere DNA-binding protein complex CBF3 subunit A CEP2 CTF14 YGR140W CBF2 CBF3A NDC10 Chromosome transmission fidelity protein 14 Kinetochore protein CTF14 Centromere-binding factor 2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MRSSILFLLKLMKIMDVQQQQEAMSSEDRFQELVDSLKPRTAHQYKTYYTKYIQWCQLNQIIPTPEDNSVNSVPYKDLPISAELIHWFLLDTLITDDKPGEKREETEDLDEEEENSFKIATLKKIIGSLNFLSKLCKVHENPNANIDTKYLESVTKLHTHWIDSQKAITTNETNNTNTQVLCPPLLKVSLNLWNPETNHLSEKFFKTCSEKLRFLVDFQLRSYLNLSFEERSKIRFGSLKLGKRDRDAIIYHKVTHSAEKKDTPGHHQLLALLPQDCPFICPQTTLAAYLYLRFYGIPSVSKGDGFPNLNADENGSLLQDIPILRGKSLTTYPREETFSNYYTTVFRYCHLPYKRREYFNKCNLVYPTWDEDTFRTFFNEENHGNWLEQPEAFAFPDKIPFDFKKIMNFKSPYTSYSTNAKKDPFPPPKDLLVQIFPEIDEYKRHDYEGLSQNSRDFLDLMEVLRERFLSNLPWIYKFFPNHDIFQDPIFGNSDFQSYFNDKTIHSKGSPILSFDILPGFNKIYKNKTNFYSLLIERPSQLTFASSHNPDTHPTQKQESEGPLQMSQLDTTQLNELLKQQSFEYVQFQTLSNFQILLSVFNKIFEKLEMKKSSRGYILHQLNLFKITLDERIKKSKIDDADKFIRDNQPIKKEENIVNEDGPNTSRRTKRPKQIRLLSIADSSDESSTEDSNVFKKDGESIEDGAYGENEDENDSEMQEQLKSMINELINSKISTFLRDQMDQFELKINALLDKILEEKVTRIIEQKLGSHTGKFSTLKRPQLYMTEEHNVGFDMEVPKKLRTSGKYAETVKDNDDHQAMSTTASPSPEQDQEAKSYTDEQEFMLDKSIDSIEGIILEWFTPNAKYANQCVHSMNKSGNKSWRANCEALYKERKSIVEFYIYLVNHESLDRYKAVDICEKLRDQNEGSFSRLAKFLRKWRHDHQNSFDGLLVYLSN C93FB7E033931E3D sgt1_yeast Protein SGT1 SGT1 YOR057W YOR29-08 Suppressor of G2 allele of SKP1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MPVEKDLKTAYKALYDEKEPLKALHLYDEILKGSPTNLTALIFKAACLEKLYFGFSDWHSDATMENAKELLDKALMTAEGRGDRSKIGLVNFRYFVHFFNIKDYELAQSYFKKAKNLGYVDDTLPLWEDRLETKLNKKNKKQKDSTNKHTIKPVESIENRGDNNSSHSPISPLKIETAPQESPKFKIDWYQSSTSVTISLFTVNLPESKEQVNIYISPNDRRTLSISYQVPKSGSEFQYNAKLSHEVDPKAVSLKIFPKKLEITLSKIDSTQWKKLEEDILTESSRLSDEGKNSDSATRLLSAETASKERLSYPSSSKKKIDWSKLDIDEEADEEAGSADSFFQKLYAGADPDTKRAMMKSFIESNGTALSTDWEDVSKGTVKTSPPEGMEPKHW 5DE65874BE155552 5DE65874BE155552 skp1_yeast Suppressor of kinetochore protein 1 SKP1 CBF3D YDR328C D9798.14 Centromere DNA-binding protein complex CBF3 subunit D E3 ubiquitin ligase complex SCF subunit SKP1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MVTSNVVLVSGEGERFTVDKKIAERSLLLKNYLNDMHDSNLQNNSDSESDSDSETNHKSKDNNNGDDDDEDDDEIVMPVPNVRSSVLQKVIEWAEHHRDSNFPDEDDDDSRKSAPVDSWDREFLKVDQEMLYEIILAANYLNIKPLLDAGCKVVAEMIRGRSPEEIRRTFNIVNDFTPEEEAAIRRENEWAEDR 746DDE6470A69432 746DDE6470A69432 cbf3c_yeast Centromere DNA-binding protein complex CBF3 subunit C CTF13 CBF3C YMR094W YM9582.19 YM6543.01 Chromosome transmission fidelity protein 13 Kinetochore protein CTF13 protein protein yeast Saccharomyces cerevisiae Baker's yeast MPSFNPVRFLELPIDIRKEVYFHLDGNFCGAHPYPIDILYKSNDVELPGKPSYKRSKRSKKLLRYMYPVFATYLNIFEYSPQLIEKWLEYAFWLRYDCLVLDCFKVNHLYDGTLIDALEWTYLDNELRLAYFNKASMLEVWYTFKEYKKWVIDSVAFDELDLLNVSNIQFNIDNLTPQLVDKCLSILEQKDLFATIGEVQFGQDEEVGEEKDVDVSGANSDENSSPSSTIKNKKRSASKRSHSDNGNVGATHNQLTSISVIRTIRSMESMKSLRKITVRGEKLYELLINFHGFRDNPGKTISYIVKRRINEIRLSRMNQISRTGLADFTRWDNLQKLVLSRVAYIDLNSIVFPKNFKSLTMKRVSKIKWWNIEENILKELKVDKRTFKSLYIKEDDSKFTKFFNLRHTRIKELDKSEINQITYLRCQAIVWLSFRTLNHIKLQNVSEVFNNIIVPRALFDSKRVEIYRCEKISQVLVI 77B1C012FD74E56F 77B1C012FD74E56F cbf3b_yeast Centromere DNA-binding protein complex CBF3 subunit B CEP3 CBF3 YMR168C YM8520.17C CBF3B CSL1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MFNRTTQLKSKHPCSVCTRRKVKCDRMIPCGNCRKRGQDSECMKSTKLITASSSKEYLPDLLLFWQNYEYWITNIGLYKTKQRDLTRTPANLDTDTEECMFWMNYLQKDQSFQLMNFAMENLGALYFGSIGDISELYLRVEQYWDRRADKNHSVDGKYWDALIWSVFTMCIYYMPVEKLAEIFSVYPLHEYLGSNKRLNWEDGMQLVMCQNFARCSLFQLKQCDFMAHPDIRLVQAYLILATTTFPYDEPLLANSLLTQCIHTFKNFHVDDFRPLLNDDPVESIAKVTLGRIFYRLCGCDYLQSGPRKPIALHTEVSSLLQHAAYLQDLPNVDVYREENSTEVLYWKIISLDRDLDQYLNKSSKPPLKTLDAIRRELDIFQYKVDSLEEDFRSNNSRFQKFIALFQISTVSWKLFKMYLIYYDTADSLLKVIHYSKVIISLIVNNFHAKSEFFNRHPMVMQTITRVVSFISFYQIFVESAAVKQLLVDLTELTANLPTIFGSKLDKLVYLTERLSKLKLLWDKVQLLDSGDSFYHPVFKILQNDIKIIELKNDEMFSLIKGLGSLVPLNKLRQESLLEEEDENNTEPSDFRTIVEEFQSEYNISDILS 2E24A0508080A09B 2E24A0508080A09B sgt1-skp1-1 1 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait Q08446:p.Thr146Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain T S Wed May 02 20:23:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr146ser' Tue Jun 30 18:27:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr146Ser' physical association physical association Fig. 1C To test whether this is the case for SKP1 mutations, we coexpressed Sgt1p with either wild-type Skp1p or Skp1-4p in insect cells, Sgt1p complexes were isolated using a polyclonal serum against Sgt1p and, as expected, Skp1p specifically coprecipitated with Sgt1p, skp1-sgt1 1 7 unspecified role unspecified role bait bait P52286:p.Leu146Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L S Wed May 02 16:57:31 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu146ser' Tue Jun 30 19:57:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu146Ser' 6 unspecified role unspecified role prey prey physical association physical association Fig. 1D ctf13-sgt1-1 2 8 unspecified role unspecified role bait bait region 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 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey association association Fig. 2A fter the isolation of GST-Ctf13p, we observed the copurification of Skp1p and low but specific levels of Sgt1p copurifying (Figure 2A) Mutating SKP1 to leu146ser, decreases Skp1 binding to the complex but increases the association between Ctf13 and Sgt1. ctf13-sgt1-2 2 8 unspecified role unspecified role bait bait region 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 6 unspecified role unspecified role prey prey physical association physical association Fig. 2A ctf13-cbf2 2 5 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait region 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 7 unspecified role unspecified role prey prey association association Fig.3B Mutating SKP1 to leu146ser, decreases Cep3 binding to the complex Active GST-Ctf13p produced in the presence of wild-type Skp1p bound specifically to both Ndc10p and Cep3p (Figure 3B) ctf13-cbf2-1 2 8 unspecified role unspecified role bait bait region 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 5 unspecified role unspecified role prey prey physical association physical association Fig.3B sgt1-skp1-4 3 6 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role ancillary ancillary region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig.5D sgt1-skp1-5 3 7 unspecified role unspecified role prey prey P52286:p.Ile172Asn mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain I N Wed May 02 15:02:16 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile172asn' Tue Jun 30 17:23:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile172Asn' P52286:p.Leu146Ser mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L S Wed May 02 15:45:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu146ser' Tue Jun 30 18:05:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu146Ser' 6 unspecified role unspecified role bait bait physical association physical association Fig.5D ctf13-cep3 4 9 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role bait bait physical association physical association Fig. 5D