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/ Cell cycle-dependent assembly of a Gin4-septin complex. Cell cycle-dependent assembly of a Gin4-septin complex. Mol. Biol. Cell (1059-1524) 2002 imex curation Mortensen EM., McDonald H., Yates J., Kellogg DR. kellogg@darwin.ucsc.edu 27-Oct-2007: Contacted by JYOTI. davet 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 Mortensen EM., McDonald H., Yates J., Kellogg DR. kellogg@darwin.ucsc.edu Accepted 2009-OCT-17 by DAVET. 27-Oct-2007: Contacted by JYOTI. Mol. Biol. Cell (1059-1524) 2002 davet Only protein-protein interactions imex curation Cell cycle-dependent assembly of a Gin4-septin complex. Cell cycle-dependent assembly of a Gin4-septin complex. Mol. Biol. Cell (1059-1524) 2002 imex curation Mortensen EM., McDonald H., Yates J., Kellogg DR. kellogg@darwin.ucsc.edu 27-Oct-2007: Contacted by JYOTI. davet 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 Mortensen EM., McDonald H., Yates J., Kellogg DR. kellogg@darwin.ucsc.edu Accepted 2009-OCT-17 by DAVET. 27-Oct-2007: Contacted by JYOTI. Mol. Biol. Cell (1059-1524) 2002 davet Only protein-protein interactions imex curation Cell cycle-dependent assembly of a Gin4-septin complex. Cell cycle-dependent assembly of a Gin4-septin complex. Mol. Biol. Cell (1059-1524) 2002 imex curation Mortensen EM., McDonald H., Yates J., Kellogg DR. kellogg@darwin.ucsc.edu 27-Oct-2007: Contacted by JYOTI. davet Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip de novo protein sequence de novo protein sequencing by mass spectrometry de novo protein sequence Mortensen EM., McDonald H., Yates J., Kellogg DR. kellogg@darwin.ucsc.edu Accepted 2009-OCT-17 by DAVET. Mol. Biol. Cell (1059-1524) 2002 davet Only protein-protein interactions imex curation Cell cycle-dependent assembly of a Gin4-septin complex. Cell cycle-dependent assembly of a Gin4-septin complex. Mol. Biol. Cell (1059-1524) 2002 imex curation Mortensen EM., McDonald H., Yates J., Kellogg DR. kellogg@darwin.ucsc.edu 27-Oct-2007: Contacted by JYOTI. davet Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Mortensen EM., McDonald H., Yates J., Kellogg DR. Purified septin complex phosphorylated by Gin4.Component proteins then separated by SDS PAGE and identity of phosphorylated protein confirmed by western blot. kellogg@darwin.ucsc.edu Accepted 2009-OCT-17 by DAVET. 27-Oct-2007: Contacted by JYOTI. Mol. Biol. Cell (1059-1524) 2002 davet Only protein-protein interactions imex curation bni5_yeast Bud neck protein 5 BNI5 N1706 YNL166C protein protein yeast Saccharomyces cerevisiae Baker's yeast MGLDQDKIKKRLSQIEIDINQMNQMIDENLQLVEPAEDEAVEDNVKDTGVVDAVKVAETALFSGNDGADSNPGDSAQVEEHKTAQVHIPTENEANKSTDDPSQLSVTQPFIAKEQITHTAIAIGDSYNSFVANSAGNEKAKDSCTENKEDGTVNIDQNRGEADVEIIENNDDEWEDEKSDVEEGRVDKGTEENSEIESFKSPMPQNNTLGGENKLDAELVLDKFSSANKDLDIQPQTIVVGGDNEYNHESSRLADQTPHDDNSENCPNRSGGSTPLDSQTKIFIPKKNSKEDGTNINHFNSDGDGQKKMANFETRRPTNPFRVISVSSNSNSRNGSRKSSLNKYDSPVSSPITSASELGSIAKLEKRHDYLSMKCIKLQKEIDYLNKMNAQGSLSMEDGKRLHRAVVKLQEYLDKKTKEKYEVGVLLSRHLRKQIDRGENGQFWIGTK 4861981BE3E15E21 gin4_yeast Serine/threonine-protein kinase GIN4 GIN4 YDR507C D9719.13 Growth inhibitory protein 4 protein protein yeast Saccharomyces cerevisiae Baker's yeast MAINGNSIPAIKDNTIGPWKLGETLGLGSTGKVQLARNGSTGQEAAVKVISKAVFNTGNVSGTSIVGSTTPDALPYGIEREIIIMKLLNHPNVLRLYDVWETNTDLYLVLEYAEKGELFNLLVERGPLPEHEAIRFFRQIIIGVSYCHALGIVHRDLKPENLLLDHKYNIKIADFGMAALETEGKLLETSCGSPHYAAPEIVSGIPYQGFASDVWSCGVILFALLTGRLPFDEEDGNIRTLLLKVQKGEFEMPSDDEISREAQDLIRKILTVDPERRIKTRDILKHPLLQKYPSIRDSKSIRGLPREDTYLTPLSESNSSIDATILQNLVILWHGRDPEGIKEKLREPGANAEKTLYALLYRFKCDTQKELIKQQQVKKRQSISSVSVSPSKKVSTTPQRRRNRESLISVTSSRKKPISFNKFTASSASSSNLTTPGSSKRLSKNFSSKKKLSTIVNQSSPTPASRNKRASVINVEKNQKRASIFSTTKKNKRSSRSIKRMSLIPSMKRESVTTKLMSTYAKLAEDDDWEYIEKETKRTSSNFATLIDEIFEYEKYEQIRKEKEELERKVREAKAREELERRRRKQEEKERARKLLEKEDLKRKQEELKKQIEIDISDLEQELSKHKEEKLDGNIRSISAPMENEEKNINHLEVDIDNILRRRNFSLQTRPVSRLDPGIMFSSPTEEVSPVEPKRTENERLTTEKKILETIRRSKFLGSSFNIDKELKLSKMEYPSIIAPQRLSEERVVSDSNDGYESLILPKDGNGVSQLKDSTATTAPVSDGRLRKISEIRVPQFTRKSRHFSESNKRLSVLSMYSTKESFTNLVDILKNGNLDVNNQQSQRIPTPRSADDSEFLFETVNEEAEYTGNSSNDERLYDVGDSTIKDKSALKLNFADRFNGSNEAKQTDNLHLPILPPLNGDNELRKQNSQEGDQAHPKIKSMIPESGSSSHTEKEEENEEKEEKKPEQHKQEEDQEKREKVVDDMEPPLNKSVQKIREKNAGSQAKDHSKDHLKEHKQDKNTAIGNGSFFRKFSKSSDKTMELYAKISAKQLFNGLEKLLRGWTQYGLKNIKSHPNNLTLTGKLSSDNIFSLRSTLFEVNIYPRGKMSVVQFKKVSGSFKAVKKLVNEVENVLNKEGVLQK EC16FF4BB49DD811 EC16FF4BB49DD811 cdc10_yeast Cell division control protein 10 CDC10 YCR2C YCR002C YCR022 protein protein yeast Saccharomyces cerevisiae Baker's yeast MDPLSSVQPASYVGFDTITNQIEHRLLKKGFQFNIMVVGQSGLGKSTLINTLFASHLIDSATGDDISALPVTKTTEMKISTHTLVEDRVRLNINVIDTPGFGDFIDNSKAWEPIVKYIKEQHSQYLRKELTAQRERFITDTRVHAILYFLQPNGKELSRLDVEALKRLTEIANVIPVIGKSDTLTLDERTEFRELIQNEFEKYNFKIYPYDSEELTDEELELNRSVRSIIPFAVVGSENEIEINGETFRGRKTRWSAINVEDINQCDFVYLREFLIRTHLQDLIETTSYIHYEGFRARQLIALKENANSRSSAHMSSNAIQR 672467301D4D2697 672467301D4D2697 cdc3_yeast Cell division control protein 3 CDC3 L8543.7 YLR314C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSLKEEQVSIKQDPEQEERQHDQFNDVQIKQESQDHDGVDSQYTNGTQNDDSERFEAAESDVKVEPGLGMGITSSQSEKGQVLPDQPEIKFIRRQINGYVGFANLPKQWHRRSIKNGFSFNLLCVGPDGIGKTTLMKTLFNNDDIEANLVKDYEEELANDQEEEEGQGEGHENQSQEQRHKVKIKSYESVIEENGVKLNLNVIDTEGFGDFLNNDQKSWDPIIKEIDSRFDQYLDAENKINRHSINDKRIHACLYFIEPTGHYLKPLDLKFMQSVYEKCNLIPVIAKSDILTDEEILSFKKTIMNQLIQSNIELFKPPIYSNDDAENSHLSERLFSSLPYAVIGSNDIVENYSGNQVRGRSYPWGVIEVDNDNHSDFNLLKNLLIKQFMEELKERTSKILYENYRSSKLAKLGIKQDNSVFKEFDPISKQQEEKTLHEAKLAKLEIEMKTVFQQKVSEKEKKLQKSETELFARHKEMKEKLTKQLKALEDKKKQLELSINSASPNVNHSPVPTKKKGFLR DCCD4C8C16AA5E25 DCCD4C8C16AA5E25 shs1_yeast Seventh homolog of septin 1 YDL225W Septation protein 7 SHS1 SEP7 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSTASTPPINLFRRKKEHKRGITYTMLLCGPAGTGKTAFANNLLETKIFPHKYQYGKSNASISSNPEVKVIAPTKVVSFNSKNGIPSYVSEFDPMRANLEPGITITSTSLELGGNKDQGKPEMNEDDTVFFNLIMTHGIGENLDDSLCSEEVMSYLEQQFDIVLAEETRIKRNPRFEDTRVHVALYFIEPTGHGLREVDVELMKSISKYTNVLPIITRADSFTKEELTQFRKNIMFDVERYNVPIYKFEVDPEDDDLESMEENQALASLQPFAIITSDTRDSEGRYVREYPWGIISIDDDKISDLKVLKNVLFGSHLQEFKDTTQNLLYENYRSEKLSSVANAEEIGPNSTKRQSNAPSLSNFASLISTGQFNSSQTLANNLRADTPRNQVSGNFKENEYEDNGEHDSAENEQEMSPVRQLGREIKQENENLIRSIKTESSPKFLNSPDLPERTKLRNISETVPYVLRHERILARQQKLEELEAQSAKELQKRIQELERKAHELKLREKLINQNKLNGSSSSINSLQQSTRSQIKKNDTYTDLASIASGRD BDA2631061A278F9 BDA2631061A278F9 cdc11_yeast Cell division control protein 11 CDC11 PSL9 YJR076C J1833 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSGIIDASSALRKRKHLKRGITFTVMIVGQSGSGRSTFINTLCGQQVVDTSTTILLPTDTSTEIDLQLREETVELEDDEGVKIQLNIIDTPGFGDSLDNSPSFEIISDYIRHQYDEILLEESRVRRNPRFKDGRVHCCLYLINPTGHGLKEIDVEFIRQLGSLVNIIPVISKSDSLTRDELKLNKKLIMEDIDRWNLPIYNFPFDEDEISDEDYETNMYLRTLLPFAIIGSNEVYEMGGDVGTIRGRKYPWGILDVEDSSISDFVILRNALLISHLHDLKNYTHEILYERYRTEALSGESVAAESIRPNLTKLNGSSSSSTTTRRNTNPFKQSNNINNDVLNPASDMHGQSTGENNETYMTREEQIRLEEERLKAFEERVQQELLLKRQELLQREKELREIEARLEKEAKIKQEE 5CFFD4CD1F6C57D4 5CFFD4CD1F6C57D4 cdc12_yeast Cell division control protein 12 CDC12 CLA10 PSL7 YHR107C Septin protein protein yeast Saccharomyces cerevisiae Baker's yeast MSAATATAAPVPPPVGISNLPNQRYKIVNEEGGTFTVMLCGESGLGKTTFINTLFQTVLKRADGQQHRQEPIRKTVEIDITRALLEEKHFELRVNVIDTPGFGDNVNNNKAWQPLVDFIDDQHDSYMRQEQQPYRTKKFDLRVHAVLYFIRPTGHGLKPIDIETMKRLSTRANLIPVIAKADTLTAQELQQFKSRIRQVIEAQEIRIFTPPLDADSKEDAKSGSNPDSAAVEHARQLIEAMPFAIVGSEKKFDNGQGTQVVARKYPWGLVEIENDSHCDFRKLRALLLRTYLLDLISTTQEMHYETYRRLRLEGHENTGEGNEDFTLPAIAPARKLSHNPRYKEEENALKKYFTDQVKAEEQRFRQWEQNIVNERIRLNGDLEEIQGKVKKLEEQVKSLQVKKSHLK A72DC393A6DBF924 A72DC393A6DBF924 nap1_yeast Nucleosome assembly protein NAP1 YKR048C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSDPIRTKPKSSMQIDNAPTPHNTPASVLNPSYLKNGNPVRAQAQEQDDKIGTINEEDILANQPLLLQSIQDRLGSLVGQDSGYVGGLPKNVKEKLLSLKTLQSELFEVEKEFQVEMFELENKFLQKYKPIWEQRSRIISGQEQPKPEQIAKGQEIVESLNETELLVDEEEKAQNDSEEEQVKGIPSFWLTALENLPIVCDTITDRDAEVLEYLQDIGLEYLTDGRPGFKLLFRFDSSANPFFTNDILCKTYFYQKELGYSGDFIYDHAEGCEISWKDNAHNVTVDLEMRKQRNKTTKQVRTIEKITPIESFFNFFDPPKIQNEDQDEELEEDLEERLALDYSIGEQLKDKLIPRAVDWFTGAALEFEFEEDEEEADEDEDEEEDDDHGLEDDDGESAEEQDDFAGRPEQAPECKQS C0F97FBA1B9EEA83 C0F97FBA1B9EEA83 gin4-cdc3-1 3 6 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey association association We found that Cdc3, Cdc10, Cdc11, Cdc12, Shs1, Nap1, and Bni5 are all retained on the anti-HA-Gin4 column but not on the anti-GST control column (Table 1). Fig. 3A, Table 1 Single peptides for Ctf7 (P43605), YJR083C (P47129), and YLL054C (Q12244) were identified my mass spec but could not be observed by Coomassie Blue staining, other proteins were present in both te control and pulldown column. gin4-nap1-1 1 6 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey association association We verified the presence of Gin4, Cdc11, Shs1, and Nap1 by Western blotting (Figure 3B), Fig. 3B gin4-nap1-2 2 12 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait association association We found that the Gin4-Cdc11 complex is detected only during mitosis, whereas Gin4 and Nap1 are bound to each other throughout the cell cycle. Figs. 4A, 6A Cells were synchronised in G1 by treatment with alpha factor, temperature shift and the addition of dextrose-containing media with no effect on results. Loss of interaction was not caused by activation of an Swe1-dependent checkpoint. gin4-nap1-3 2 12 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait association association We found that the Gin4-Cdc11 complex is detected only during mitosis, whereas Gin4 and Nap1 are bound to each other throughout the cell cycle. Fig. 4A, 4C, 5B, 6A Interaction is not cell-cycle regulated and does not require Gin4 hyperphosphorylation. gin4-nap1-4 2 6 unspecified role unspecified role bait bait region (required) phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey association association Gin4 was immunoprecipitated (IP) from the indicated samples and coprecipitation of Nap1, Cdc11, and Shs1 was monitored by Western blotting. Fig. 5B 1NM-PP1 - an inhibitor of CDC28 gin4-shs1-1 2 6 unspecified role unspecified role bait bait Q12263:p.Lys48Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K A Wed May 02 18:33:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys48ala' Tue Jun 30 19:10:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys48Ala' 9 unspecified role unspecified role prey prey physical association physical association We found that a fraction of Shs1 is able to interact with Gin4 independently of Cdc11 in all backgrounds tested, with the exception of the gin4K48A allele (Figure 6C). Fig. 6C shs1-cdc3-1 3 8 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association We found that Shs1 copurifies with Cdc3, Cdc10, Cdc11, and Cdc12 as determined by Western blotting and mass spectrometry Fig. 8A Column washed with 1M KCl gin4-shs1-2 4 6 enzyme enzyme neutral component neutral component Q12263:p.Lys48Ala Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K A Wed May 02 15:12:31 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys48ala' Tue Jun 30 19:48:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys48Ala' yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 9 enzyme target enzyme target substrate substrate neutral component neutral component yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast phosphorylation phosphorylation reaction phosphorylation Figs. 8B and 8C These experiments suggest that Gin4 directly phosphorylates Shs1 in vitro.