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/ http://www.ebi.ac.uk/ hirohashi-2006-2 Centrosomal proteins Nde1 and Su48 form a complex regulated by phosphorylation. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Hirohashi Y., Wang Q., Liu Q., Li B., Du X., Zhang H., Furuuchi K., Masuda K., Sato N., Greene MI. Apoptosis - Interactions involving proteins with a function related to apoptosis A deleted form of the Su48 protein was expressed in 293T cells the authors contest that the deleted form appears to be more stable, probably due to removal of the Destruction Box (located within amino acid residues 316 - 407 of the protein). Figure 1 (a). Figure 2.Figure 3. greene@reo.med.upenn.edu Human Hela cell line cDNA library Oncogene (0950-9232) 2006 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer hirohashi-2006-4 Centrosomal proteins Nde1 and Su48 form a complex regulated by phosphorylation. in vitro In vitro in gel kinase assay in-gel kinase assay in gel kinase assay predetermined predetermined participant predetermined Hirohashi Y., Wang Q., Liu Q., Li B., Du X., Zhang H., Furuuchi K., Masuda K., Sato N., Greene MI. Apoptosis - Interactions involving proteins with a function related to apoptosis Figure 4 (b). greene@reo.med.upenn.edu Human Hela cell line cDNA library Oncogene (0950-9232) 2006 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer hirohashi-2006-1 Centrosomal proteins Nde1 and Su48 form a complex regulated by phosphorylation. 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Hirohashi Y., Wang Q., Liu Q., Li B., Du X., Zhang H., Furuuchi K., Masuda K., Sato N., Greene MI. Apoptosis - Interactions involving proteins with a function related to apoptosis greene@reo.med.upenn.edu Human Hela cell line cDNA library Oncogene (0950-9232) 2006 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer hirohashi-2006-3 Centrosomal proteins Nde1 and Su48 form a complex regulated by phosphorylation. human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference fluorescence imaging fluorescence microscopy fluorescence imaging predetermined predetermined participant predetermined Hirohashi Y., Wang Q., Liu Q., Li B., Du X., Zhang H., Furuuchi K., Masuda K., Sato N., Greene MI. Apoptosis - Interactions involving proteins with a function related to apoptosis A positive control using RFP-Nde1 and gamma tubulin was also carried out to authenticate centrosomal colocalisation of the proteins studied. However it was not possible to identify the isoform of gamma tubulin used in the study. A deleted form of the Su48 protein was expressed in 293T cells the authors contest that the deleted form appears to be more stable, probably due to removal of the Destruction Box (located within amino acid residues 316 - 407 of the protein). Figure 1 (b) to (e). Figure 2. Figure 5. greene@reo.med.upenn.edu Human Hela cell line cDNA library Oncogene (0950-9232) 2006 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer q9nxr1-1 Nuclear distribution protein nudE homolog 1 NDE1 NUDE protein protein human Homo sapiens Human MEDSGKTFSSEEEEANYWKDLAMTYKQRAENTQEELREFQEGSREYEAELETQLQQIETRNRDLLSENNRLRMELETIKEKFEVQHSEGYRQISALEDDLAQTKAIKDQLQKYIRELEQANDDLERAKRATIMSLEDFEQRLNQAIERNAFLESELDEKENLLESVQRLKDEARDLRQELAVQQKQEKPRTPMPSSVEAERTDTAVQATGSVPSTPIAHRGPSSSLNTPGSFRRGLDDSTGGTPLTPAARISALNIVGDLLRKVGALESKLASCRNLVYDQSPNRTGGPASGRSSKNRDGGERRPSSTSVPLGDKGLGKRLEFGKPPSHMSSSPLPSAQGVVKMLL zn365_human Protein ZNF365 DBZ DISC1-binding zinc-finger protein ZNF365 KIAA0844 Protein su48 protein protein human Homo sapiens Human MQQKAFEESRYPWQESFENVAVCLPLRCPRCGDHTRFRSLSSLRAHLEFSHSYEERTLLTKCSLFPSLKDTDLVTSSELLKPGKLQSSGNVVKQKPSYVNLYSISHEHSKDRKPFEVVAERPVSYVQTYTAMDLHADSLDGTRSGPGLPTSDTKASFEAHVREKFNRMVEAVDRTIEKRIDKLTKELAQKTAELLEVRAAFVQLTQKKQEVQRRERALNRQVDVAVEMIAVLRQRLTESEEELLRKEEEVVTFNHFLEAAAEKEVQGKARLQDFIENLLQRVELAEKQLEYYQSQQASGFVRDLSGHVLTDISSNRKPKCLSRGHPHSVCNHPDLKAHFHPKGRNHLKKAKDDRASMQPAKAIHEQAESSRDLCRPPKKGELLGFGRKGNIRPKMAKKKPTAIVNII ndel1_human Nuclear distribution protein nudE-like 1 NDEL1 Mitosin-associated protein 1 EOPA MITAP1 NUDEL protein protein human Homo sapiens Human MDGEDIPDFSSLKEETAYWKELSLKYKQSFQEARDELVEFQEGSRELEAELEAQLVQAEQRNRDLQADNQRLKYEVEALKEKLEHQYAQSYKQVSVLEDDLSQTRAIKEQLHKYVRELEQANDDLERAKRATIVSLEDFEQRLNQAIERNAFLESELDEKESLLVSVQRLKDEARDLRQELAVRERQQEVTRKSAPSSPTLDCEKMDSAVQASLSLPATPVGKGTENTFPSPKAIPNGFGTSPLTPSARISALNIVGDLLRKVGALESKLAACRNFAKDQASRKSYISGNVNCGVLNGNGTKFSRSGHTSFFDKGAVNGFDPAPPPPGLGSSRPSSAPGMLPLSV nde1_human Nuclear distribution protein nudE homolog 1 NDE1 NUDE protein protein human Homo sapiens Human MEDSGKTFSSEEEEANYWKDLAMTYKQRAENTQEELREFQEGSREYEAELETQLQQIETRNRDLLSENNRLRMELETIKEKFEVQHSEGYRQISALEDDLAQTKAIKDQLQKYIRELEQANDDLERAKRATIMSLEDFEQRLNQAIERNAFLESELDEKENLLESVQRLKDEARDLRQELAVQQKQEKPRTPMPSSVEAERTDTAVQATGSVPSTPIAHRGPSSSLNTPGSFRRGLDDSTGGTPLTPAARISALNIVGDLLRKVGALESKLASCRNLVYDQSPNRTGGPASGRSSKNRDGGERRPSSTSVPLGDKGLDTSCRWLSKSTTRSSSSC cdk1_human Cyclin-dependent kinase 1 p34 protein kinase Cell division protein kinase 1 CDK1 CDC2 Cell division control protein 2 homolog CDC28A CDKN1 P34CDC2 protein protein human Homo sapiens Human MEDYTKIEKIGEGTYGVVYKGRHKTTGQVVAMKKIRLESEEEGVPSTAIREISLLKELRHPNIVSLQDVLMQDSRLYLIFEFLSMDLKKYLDSIPPGQYMDSSLVKSYLYQILQGIVFCHSRRVLHRDLKPQNLLIDDKGTIKLADFGLARAFGIPIRVYTHEVVTLWYRSPEVLLGSARYSTPVDIWSIGTIFAELATKKPLFHGDSEIDQLFRIFRALGTPNNEVWPEVESLQDYKNTFPKWKPGSLASHVKNLDENGLDLLSKMLIYDPAKRISGKMALNHPYFNDLDNQIKKM su48-nde1-1 Coimmunoprecipitation analysis reveals interaction between Su48 protein and NDe1 protein 377361 n/a 377365 unspecified role unspecified role bait bait region 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 false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 377366 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 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 su48-ndel1 Coimmunoprecipitation analysis reveals interaction between Su48 protein and Ndel1 protein 377361 N/A 377366 unspecified role unspecified role prey prey coiled coil region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 377367 unspecified role unspecified role bait bait region 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 false physical association physical association false false false su48-nde1-3 Coimmunoprecipitation analysis reveals interaction between Su48 protein and NDe1 protein 377361 N/A 377366 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 377368 unspecified role unspecified role bait bait region 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 false Q9NXR1:p.[Thr191Glu;Thr215Glu;Thr228Glu;Thr243Glu;Thr246Glu;Ser282Glu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Interaction was shown to be abolished when residues 191, 215, 228, 243, 246, 282 (all the potential phosphorylation sites) were replaced with glutamic acid residues, which mimics the effects of phosphorylation. cdc2-nde1 Kinase analysis reveals NDe1 protein is phosphorylated by the Cdc2 protein 377362 N/A 377368 enzyme target enzyme target substrate substrate unspecified role unspecified role region 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 false Q9NXR1:p.[Thr191Val;Thr215Val;Thr228Val;Thr243Val;Thr246Val;Ser282Val] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false N/A 377369 enzyme enzyme unspecified role unspecified role phosphorylation phosphorylation reaction phosphorylation false false false Deletion of the C terminus of Nde1 (amino acid residues 134 to 335)abolished phosphorylation. Note all putative phosphorylation sites are within the C terminus region. Phosphorylation of NDE1 cdc2-ndel1 Kinase analysis reveals Ndel1 protein is phosphorylated by the Cdc2 protein 377362 N/A 377369 enzyme enzyme unspecified role unspecified role N/A 377367 enzyme target enzyme target substrate substrate unspecified role unspecified role region 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 false phosphorylation phosphorylation reaction phosphorylation false false false Phosphorylation of NDE1 su48-nde1 Yeast 2 hybrid analysis reveals interaction between Su48 protein and NDe1 protein 377363 N/A 377368 unspecified role unspecified role prey prey N/A 377366 unspecified role unspecified role bait bait physical association physical association false false false su48-nde1-2 Colocalisation analysis reveals interaction between Su48 protein and NDe1 protein 377364 N/A 377368 unspecified role unspecified role neutral component neutral component region red florescent protein tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Q9NXR1:p.[Thr191Glu;Thr215Glu;Thr228Glu;Thr243Glu;Thr246Glu;Ser282Glu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false N/A 377366 unspecified role unspecified role neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false The proteins colocalised to the centrosomal regions The authors state "It is noteworthy that GFP-Nde1 is targeted to the centrosomal dots during the interphase or S-phase, but not in the M-phase". The region 184 to 335 amino acid residues was required for Nde1 localisation to the centrosome. Additionally it was demonstrated that colocalisation was abolished when putative phosphorylation sites at amino acid residues 191, 215, 228, 243, 246 and 282 were phosphorylated.