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/ The amino-terminal portion of the F-box protein Met30p mediates its nuclear import and assimilation into an SCF complex. The amino-terminal portion of the F-box protein Met30p mediates its nuclear import and assimilation into an SCF complex. J. Biol. Chem. (0021-9258) 2004 imex curation Brunson LE., Dixon C., Kozubowski L., Mathias N. Mutations were made within and upstream of the F-box of the Met30 n-terminus region to determine their affect on binding with Cbf3d (Fig.3). Mapping of homomultimerization and Cbf3d binding domains in Met30 (Fig.4). 3B, 4B pmathi@lsuhsc.edu 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 Brunson LE., Dixon C., Kozubowski L., Mathias N. Mutations were made within and upstream of the F-box of the Met30 n-terminus region to determine their affect on binding with Cbf3d (Fig.3). Mapping of homomultimerization and Cbf3d binding domains in Met30 (Fig.4). 3B, 4B pmathi@lsuhsc.edu Accepted 2006-MAR-16 by ORCHARD. J. Biol. Chem. (0021-9258) 2004 Only protein-protein interactions imex curation The amino-terminal portion of the F-box protein Met30p mediates its nuclear import and assimilation into an SCF complex. The amino-terminal portion of the F-box protein Met30p mediates its nuclear import and assimilation into an SCF complex. J. Biol. Chem. (0021-9258) 2004 imex curation Brunson LE., Dixon C., Kozubowski L., Mathias N. Mutations were made within and upstream of the F-box of the Met30 n-terminus region to determine their affect on binding with Cbf3d (Fig.3). Mapping of homomultimerization and Cbf3d binding domains in Met30 (Fig.4). 3B, 4B pmathi@lsuhsc.edu Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Brunson LE., Dixon C., Kozubowski L., Mathias N. Mutations were made within and upstream of the F-box of the Met30 n-terminus region to determine their affect on binding with Cbf3d (Fig.3). Mapping of homomultimerization and Cbf3d binding domains in Met30 (Fig.4). 3B, 4B pmathi@lsuhsc.edu Accepted 2006-MAR-16 by ORCHARD. J. Biol. Chem. (0021-9258) 2004 Only protein-protein interactions imex curation The amino-terminal portion of the F-box protein Met30p mediates its nuclear import and assimilation into an SCF complex. The amino-terminal portion of the F-box protein Met30p mediates its nuclear import and assimilation into an SCF complex. J. Biol. Chem. (0021-9258) 2004 imex curation Brunson LE., Dixon C., Kozubowski L., Mathias N. Mutations were made within and upstream of the F-box of the Met30 n-terminus region to determine their affect on binding with Cbf3d (Fig.3). Mapping of homomultimerization and Cbf3d binding domains in Met30 (Fig.4). 3B, 4B pmathi@lsuhsc.edu Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast pull down pull down western blot western blot Immuno blot Immuno blot Brunson LE., Dixon C., Kozubowski L., Mathias N. Mutations were made within and upstream of the F-box of the Met30 n-terminus region to determine their affect on binding with Cbf3d (Fig.3). Mapping of homomultimerization and Cbf3d binding domains in Met30 (Fig.4). 3B, 4B pmathi@lsuhsc.edu Accepted 2006-MAR-16 by ORCHARD. J. Biol. Chem. (0021-9258) 2004 Only protein-protein interactions imex curation met30_yeast F-box protein MET30 YIL046W MET30 Methionine-requiring protein 30 E3 ubiquitin ligase complex SCF(Met30) subunit MET30 protein protein yeast Saccharomyces cerevisiae Baker's yeast MRRERQRMMSFEDKDKDDLDNSNSNNSSEMTDTAMMPPLKRLLITGSSDDLAQGSSGKKKMTMATRSPSSSPDLATNDSGTRVQPLPEYNFTKFCYRHNPDIQFSPTHTACYKQDLKRTQEINANIAKLPLQEQSDIHHIISKYSNSNDKIRKLILDGILSTSCFPQLSYISSLVTHMIKIDFISILPQELSLKILSYLDCQSLCNATRVCRKWQKLADDDRVWYHMCEQHIDRKCPNCGWGLPLLHMKRARIQQNSTGSSSNADIQTQTTRPWKVIYRERFKVESNWRKGHCRIQEFKGHMDGVLTLQFNYRLLFTGSYDSTIGIWDLFTGKLIRRLSGHSDGVKTLYFDDRKLITGSLDKTIRVWNYITGECISTYRGHSDSVLSVDSYQKVIVSGSADKTVKVWHVESRTCYTLRGHTEWVNCVKLHPKSFSCFSCSDDTTIRMWDIRTNSCLKVFRGHVGQVQKIIPLTIKDVENLATDNTSDGSSPQDDPTMTDGADESDTPSNEQETVLDENIPYPTHLLSCGLDNTIKLWDVKTGKCIRTQFGHVEGVWDIAADNFRIISGSHDGSIKVWDLQSGKCMHTFNGRRLQRETQHTQTQSLGDKVAPIACVCIGDSECFSGDEFGCVKMYKFDLND 5135D4BCA2E1EB97 5135D4BCA2E1EB97 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 met30-cbf3d-2 1 5 unspecified role unspecified role prey prey yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 4 unspecified role unspecified role bait bait n-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain P39014:p.Ser163Asn ser163asn mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S N Wed May 02 15:06:52 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser163asn' Tue Jun 30 20:12:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser163Asn' 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 P39014:p.Gln167Arg mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Q R Wed May 02 16:10:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln167arg' Tue Jun 30 19:29:19 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln167Arg' P39014:p.Ser169Pro mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain S P Wed May 02 20:19:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser169pro' Tue Jun 30 18:26:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser169Pro' P39014:p.Pro188Ser mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain P S Wed May 02 16:42:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro188ser' Tue Jun 30 18:50:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro188Ser' P39014:p.Cys211Tyr mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain C Y Wed May 02 14:26:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys211tyr' Tue Jun 30 16:51:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys211Tyr' P39014:p.Pro188Leu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain P L Wed May 02 18:53:52 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro188leu' Tue Jun 30 19:43:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro188Leu' yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association 3A Mutations within and upstream of the Met30 F-box inhibited interaction of the Met30 n-terminus with Cbf3d. met30-cbf3d-3 3 5 unspecified role unspecified role prey prey yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 4 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 P39014:p.[Leu187Asp;Glu190Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L D certain certain sequence position certain certain certain sequence position certain E A Wed May 02 14:54:21 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu187asp,glu190ala' Tue Jun 30 17:41:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu187Asp;Glu190Ala]' region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Mutations within highly conserved residues of the F-box motif in Met30 inhibited interaction of the full-length Met30 with Cbf3d. met30-cbf3d-4 2 5 unspecified role unspecified role prey prey yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 4 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain P39014:p.[Leu187Asp;Glu190Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L D certain certain sequence position certain certain certain sequence position certain E A Wed May 02 18:52:52 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu187asp,glu190ala' Tue Jun 30 17:08:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu187Asp;Glu190Ala]' yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association A region containing AA 161-169 was found to be necessary for interaction of Met30 with Cbf3d. A region containing the F-box only (AA180-225) was capable of interacting with Cbf3d as was the N-terminus region (AA 1-225). Mutations within highly conserved residues of the F-box motif in Met30 inhibited interaction of the full-length Met30 with Cbf3d. met30-met30-1 1 4 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 5 unspecified role unspecified role prey prey yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 4 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 region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast association association Residues 180-277 (which contains the F-box motif) in Met30 are required for interaction with the full-length Met30 (homomultimerization) and with Cbf3d. met30-met30-2 3 4 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 4 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 region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Residues 180-225 in Met30 are required for interaction with the full-length Met30 (homomultimerization). Point mutations within the F-box of Met30 failed to disrupt homomultimerization.