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/ jeyaprakash-2007-2 Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Jeyaprakash AA., Klein UR., Lindner D., Ebert J., Nigg EA., Conti E. Apoptosis - Interactions involving proteins with a function related to apoptosis conti@embl.de Cell (0092-8674) 2007 Only protein-protein interactions 13 imex curation The region of Borealin remaining bound to Survivin following tryptic digest was identified by mass spectrometry and N-terminal sequencing jeyaprakash-2007-3 Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. 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 Jeyaprakash AA., Klein UR., Lindner D., Ebert J., Nigg EA., Conti E. Apoptosis - Interactions involving proteins with a function related to apoptosis conti@embl.de Cell (0092-8674) 2007 Only protein-protein interactions 13 imex curation The region of Borealin remaining bound to Survivin following tryptic digest was identified by mass spectrometry and N-terminal sequencing jeyaprakash-2007-4 Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight identificat confirmation by molecular weight weight identificat Jeyaprakash AA., Klein UR., Lindner D., Ebert J., Nigg EA., Conti E. Apoptosis - Interactions involving proteins with a function related to apoptosis conti@embl.de Cell (0092-8674) 2007 Only protein-protein interactions 13 imex curation The region of Borealin remaining bound to Survivin following tryptic digest was identified by mass spectrometry and N-terminal sequencing jeyaprakash-2007-5 Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati weight identificat confirmation by molecular weight weight identificat Jeyaprakash AA., Klein UR., Lindner D., Ebert J., Nigg EA., Conti E. Apoptosis - Interactions involving proteins with a function related to apoptosis conti@embl.de Cell (0092-8674) 2007 Only protein-protein interactions 13 imex curation The region of Borealin remaining bound to Survivin following tryptic digest was identified by mass spectrometry and N-terminal sequencing jeyaprakash-2007-1 Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. in vitro In vitro footprinting footprinting ms participant Identification by mass spectrometry ms participant Jeyaprakash AA., Klein UR., Lindner D., Ebert J., Nigg EA., Conti E. Apoptosis - Interactions involving proteins with a function related to apoptosis The region of Borealin remaining bound to Survivin following tryptic digest was identified by mass spectrometry and N-terminal sequencing conti@embl.de Cell (0092-8674) 2007 Only protein-protein interactions 13 imex curation jeyaprakash-2007-6 Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Jeyaprakash AA., Klein UR., Lindner D., Ebert J., Nigg EA., Conti E. Apoptosis - Interactions involving proteins with a function related to apoptosis conti@embl.de Cell (0092-8674) 2007 Only protein-protein interactions 13 imex curation The region of Borealin remaining bound to Survivin following tryptic digest was identified by mass spectrometry and N-terminal sequencing aurkb_human Aurora kinase B AIRK2 Aurora- and IPL1-like midbody-associated protein 1 Aurora/IPL1-related kinase 2 STK-1 Serine/threonine-protein kinase 12 Serine/threonine-protein kinase 5 STK1 STK5 Aurora 1 AURKB AIK2 AIM1 ARK2 STK12 Serine/threonine-protein kinase aurora-B protein protein human Homo sapiens Human MAQKENSYPWPYGRQTAPSGLSTLPQRVLRKEPVTPSALVLMSRSNVQPTAAPGQKVMENSSGTPDILTRHFTIDDFEIGRPLGKGKFGNVYLAREKKSHFIVALKVLFKSQIEKEGVEHQLRREIEIQAHLHHPNILRLYNYFYDRRRIYLILEYAPRGELYKELQKSCTFDEQRTATIMEELADALMYCHGKKVIHRDIKPENLLLGLKGELKIADFGWSVHAPSLRRKTMCGTLDYLPPEMIEGRMHNEKVDLWCIGVLCYELLVGNPPFESASHNETYRRIVKVDLKFPASVPMGAQDLISKLLRHNPSERLPLAQVSAHPWVRANSRRVLPPSALQSVA ince_human Inner centromere protein INCENP protein protein human Homo sapiens Human MGTTAPGPIHLLELCDQKLMEFLCNMDNKDLVWLEEIQEEAERMFTREFSKEPELMPKTPSQKNRRKKRRISYVQDENRDPIRRRLSRRKSRSSQLSSRRLRSKDSVEKLATVVGENGSVLRRVTRAAAAAAAATMALAAPSSPTPESPTMLTKKPEDNHTQCQLVPVVEIGISERQNAEQHVTQLMSTEPLPRTLSPTPASATAPTSQGIPTSDEESTPKKSKARILESITVSSLMATPQDPKGQGVGTGRSASKLRIAQVSPGPRDSPAFPDSPWRERVLAPILPDNFSTPTGSRTDSQSVRHSPIAPSSPSPQVLAQKYSLVAKQESVVRRASRRLAKKTAEEPAASGRIICHSYLERLLNVEVPQKVGSEQKEPPEEAEPVAAAEPEVPENNGNNSWPHNDTEIANSTPNPKPAASSPETPSAGQQEAKTDQADGPREPPQSARRKRSYKQAVSELDEEQHLEDEELQPPRSKTPSSPCPASKVVRPLRTFLHTVQRNQMLMTPTSAPRSVMKSFIKRNTPLRMDPKCSFVEKERQRLENLRRKEEAEQLRRQKVEEDKRRRLEEVKLKREERLRKVLQARERVEQMKEEKKKQIEQKFAQIDEKTEKAKEERLAEEKAKKKAAAKKMEEVEARRKQEEEARRLRWLQQEEEERRHQELLQKKKEEEQERLRKAAEAKRLAEQREQERREQERREQERREQERREQERREQERQLAEQERRREQERLQAERELQEREKALRLQKEQLQRELEEKKKKEEQQRLAERQLQEEQEKKAKEAAGASKALNVTVDVQSPACTSYQMTPQGHRAPPKINPDNYGMDLNSDDSTDDEAHPRKPIPTWARGTPLSQAIIHQYYHPPNLLELFGTILPLDLEDIFKKSKPRYHKRTSSAVWNSPPLQGARVPSSLAYSLKKH borea_human Borealin Dasra-B Cell division cycle-associated protein 8 Pluripotent embryonic stem cell-related gene 3 protein CDCA8 PESCRG3 protein protein human Homo sapiens Human MAPRKGSSRVAKTNSLRRRKLASFLKDFDREVEIRIKQIESDRQNLLKEVDNLYNIEILRLPKALREMNWLDYFALGGNKQALEEAATADLDITEINKLTAEAIQTPLKSAKTRKVIQVDEMIVEEEEEEENERKNLQTARVKRCPPSKKRTQSIQGKGKGKRSSRANTVTPAVGRLEVSMVKPTPGLTPRFDSRVFKTPGLRTPAAGERIYNISGNGSPLADSKEIFLTVPVGGGESLRLLASDLQRHSIAQLDPEALGNIKKLSNRLAQICSSIRTHK birc5_human Baculoviral IAP repeat-containing protein 5 Apoptosis inhibitor survivin Apoptosis inhibitor 4 BIRC5 API4 IAP4 protein protein human Homo sapiens Human MGAPTLPPAWQPFLKDHRISTFKNWPFLEGCACTPERMAEAGFIHCPTENEPDLAQCFFCFKELEGWEPDDDPIEEHKKHSSGCAFLSVKKQFEELTLGEFLKLDRERAKNKIAKETNNKKKEFEETAKKVRRAIEQLAAMD birc5-cdca8-2 Interaction between BIRC5, INCENP, AURKB and CDCA8 demonstrated by coimmunoprecipitation 678936 N/A 678942 unspecified role unspecified role prey prey N/A 678943 unspecified role unspecified role prey prey N/A 678944 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 678945 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) association association false false false Figure 1F. 5D aurkb-cdca8-1 Interaction between AURKB, INCENP and CDCA8 demonstrated by coimmunoprecipitation 678936 N/A 678943 unspecified role unspecified role prey prey N/A 678942 unspecified role unspecified role prey prey N/A 678944 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false association association false false false Figure 1F, 5D Mutation of CDCA8 at W70E,F74E has no effect on this interaction but prevents binding of BIRC5 birc5-cdca8-9 Interaction between BIRC5 and CDCA8 demonstrated by coimmunoprecipitation 678936 N/A 678945 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) N/A 678944 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 5D The mutation of CDCA8 R17/19E,K20E had no effect on this interaction but prevents binding of AURKB and INCENP birc5-cdca8-10 Interaction between BIRC5, INCENP, AURKB and CDCA8 demonstrated by coimmunoprecipitation 678936 N/A 678944 unspecified role unspecified role prey prey N/A 678942 unspecified role unspecified role prey prey N/A 678945 unspecified role unspecified role prey prey N/A 678943 unspecified role unspecified role bait bait 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 association association false false false Figure 6B birc5-cdca8-11 Interaction between INCENPand AURKB demonstrated by coimmunoprecipitation 678936 N/A 678943 unspecified role unspecified role bait bait 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 678942 unspecified role unspecified role prey prey physical association physical association false false false Figure 6B Mutation of INCENP F22R,L34R had no effect on thi interaction but prevented binding to BIRC5 and CDCA8. birc5-cdca8-3 Interaction between BIRC5 and CDCA8 demonstrated by y2H 678937 N/A 678945 unspecified role unspecified role bait bait N/A 678944 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false 2A incenp-cdca8-1 Interaction between INCENP and CDCA8 demonstrated by y2H 678937 N/A 678943 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false N/A 678944 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false 678982 678980 physical association physical association false false false Figure 2A birc5-cdca8-4 Interaction between BIRC5 and CDCA8 demonstrated by size exclusion 678938 N/A 678944 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) N/A 678945 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false Figure 2C birc5-cdca8-5 Interaction between BIRC5, INCENP and CDCA8 demonstrated by size exclusion 678938 N/A 678944 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) N/A 678945 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) N/A 678943 unspecified role unspecified role neutral component neutral component n-terminal region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false Figure 2C birc5-cdca8-6 Interaction between BIRC5 and CDCA8 demonstrated by solution sedimentation 678939 N/A 678945 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) Stoichiometry: 2.0 N/A 678944 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) Stoichiometry: 2.0 direct interaction direct interaction false false false Figure 2D 3.3 s dimer of molecular weight of 53 kDa birc5-cdca8-7 Interaction between BIRC5, INCENP and CDCA8 demonstrated by size exclusion 678939 N/A 678945 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) Stoichiometry: 1.0 N/A 678943 unspecified role unspecified role neutral component neutral component n-terminal region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) Stoichiometry: 1.0 N/A 678944 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) Stoichiometry: 1.0 direct interaction direct interaction false false false Figure 2D 3.0 s monomer of 35 kDa birc5-cdca8-1 Interaction between BIRC5 and CDCA8 demonstrated by footprinting 678940 N/A 678944 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) N/A 678945 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false birc5-cdca8-8 Interaction between BIRC5, INCENP and CDCA8 demonstrated by x-ray crystallography 678941 N/A 678944 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) Stoichiometry: 1.0 N/A 678945 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) Stoichiometry: 1.0 N/A 678943 unspecified role unspecified role neutral component neutral component n-terminal region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) Stoichiometry: 1.0 physical association physical association false false false Figure 3A 1.4A 18.5% bound, 20.2% free, Borealin and INCENP bind Survivin to form a three-helix bundle. INCENP forms a single alpha-helix that is 10 turns long (residues 8-46) and stacks parallel to the Survivin C helix. Borealin is also characterized by a long alpha-helix (residues 15-60), which stacks with 12 turns against the Survivin C helix in an antiparallel fashion.