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/ Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. Cell (0092-8674) 2007 imex curation 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 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions 13 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 Accepted 2007-DEC-21 by DAVET. 22-Jan-2008: Contacted by JYOTI. Cell (0092-8674) 2007 DAVET Only protein-protein interactions 13 imex curation Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. Cell (0092-8674) 2007 imex curation 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 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions 13 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 Accepted 2007-DEC-21 by DAVET. 22-Jan-2008: Contacted by JYOTI. Cell (0092-8674) 2007 DAVET Only protein-protein interactions 13 imex curation Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. Cell (0092-8674) 2007 imex curation 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 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions 13 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 Accepted 2007-DEC-21 by DAVET. Cell (0092-8674) 2007 DAVET Only protein-protein interactions 13 imex curation Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. Cell (0092-8674) 2007 imex curation 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 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions 13 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 Accepted 2007-DEC-21 by DAVET. 22-Jan-2008: Contacted by JYOTI. Cell (0092-8674) 2007 DAVET Only protein-protein interactions 13 imex curation Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. Cell (0092-8674) 2007 imex curation 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 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions 13 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 Accepted 2007-DEC-21 by DAVET. 22-Jan-2008: Contacted by JYOTI. Cell (0092-8674) 2007 DAVET Only protein-protein interactions 13 imex curation Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. Structure of a Survivin-Borealin-INCENP core complex reveals how chromosomal passengers travel together. Cell (0092-8674) 2007 imex curation 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 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions 13 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 Accepted 2007-DEC-21 by DAVET. 22-Jan-2008: Contacted by JYOTI. Cell (0092-8674) 2007 DAVET Only protein-protein interactions 13 imex curation birc5_human Baculoviral IAP repeat-containing protein 5 Apoptosis inhibitor survivin Apoptosis inhibitor 4 BIRC5 API4 IAP4 protein protein human Homo sapiens Human MGAPTLPPAWQPFLKDHRISTFKNWPFLEGCACTPERMAEAGFIHCPTENEPDLAQCFFCFKELEGWEPDDDPIEEHKKHSSGCAFLSVKKQFEELTLGEFLKLDRERAKNKIAKETNNKKKEFEETAKKVRRAIEQLAAMD DIP protein O15392 original sequence version: 129 DIP protein O15392 original sequence version: 132 DIP protein O15392-1 original sequence version: 141 DIP protein O15392-1 original sequence version: 143 9E7CADCDF2822286 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 DIP protein Q96GD4 original sequence version: 88 DIP protein Q96GD4 original sequence version: 113 DIP protein Q96GD4 original sequence version: 116 DIP protein Q96GD4 original sequence version: 122 DIP protein Q96GD4 original sequence version: 177 DIP protein Q96GD4-1 original sequence version: 177 A5ED13EF5A1FAFBF 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 DIP protein Q53HL2 original sequence version: 57 519978A7C295C571 ince_human Inner centromere protein INCENP protein protein human Homo sapiens Human MGTTAPGPIHLLELCDQKLMEFLCNMDNKDLVWLEEIQEEAERMFTREFSKEPELMPKTPSQKNRRKKRRISYVQDENRDPIRRRLSRRKSRSSQLSSRRLRSKDSVEKLATVVGENGSVLRRVTRAAAAAAAATMALAAPSSPTPESPTMLTKKPEDNHTQCQLVPVVEIGISERQNAEQHVTQLMSTEPLPRTLSPTPASATAPTSQGIPTSDEESTPKKSKARILESITVSSLMATPQDPKGQGVGTGRSASKLRIAQVSPGPRDSPAFPDSPWRERVLAPILPDNFSTPTGSRTDSQSVRHSPIAPSSPSPQVLAQKYSLVAKQESVVRRASRRLAKKTAEEPAASGRIICHSYLERLLNVEVPQKVGSEQKEPPEEAEPVAAAEPEVPENNGNNSWPHNDTEIANSTPNPKPAASSPETPSAGQQEAKTDQADGPREPPQSARRKRSYKQAVSELDEEQHLEDEELQPPRSKTPSSPCPASKVVRPLRTFLHTVQRNQMLMTPTSAPRSVMKSFIKRNTPLRMDPKCSFVEKERQRLENLRRKEEAEQLRRQKVEEDKRRRLEEVKLKREERLRKVLQARERVEQMKEEKKKQIEQKFAQIDEKTEKAKEERLAEEKAKKKAAAKKMEEVEARRKQEEEARRLRWLQQEEEERRHQELLQKKKEEEQERLRKAAEAKRLAEQREQERREQERREQERREQERREQERREQERQLAEQERRREQERLQAERELQEREKALRLQKEQLQRELEEKKKKEEQQRLAERQLQEEQEKKAKEAAGASKALNVTVDVQSPACTSYQMTPQGHRAPPKINPDNYGMDLNSDDSTDDEAHPRKPIPTWARGTPLSQAIIHQYYHPPNLLELFGTILPLDLEDIFKKSKPRYHKRTSSAVWNSPPLQGARVPSSLAYSLKKH DIP protein Q9NQS7 original sequence version: 83 DIP protein Q9NQS7 original sequence version: 86 644D081DCC9035E8 birc5-cdca8-1 3 9 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 ecobb Escherichia coli (strain B / BL21) 7 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Upon trypsin treatment, Survivin remained intact while Borealin was digested to a stable fragment encompassing residues 10-109 (mass spectrometry and N-terminal sequencing; results not shown). birc5-cdca8-2 1 8 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 9 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 region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) association association Borealin10�109 (but not Borealin10�57) interacts with INCENP, Aurora B, and Survivin Figure 1F. 5D birc5-cdca8-3 4 7 unspecified role unspecified role bait bait 9 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 physical association physical association Yeast two-hybrid assays show the interaction of Borealin10�109 with Survivin 2A incenp-cdca8-1 4 10 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 9 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 28 30 physical association physical association Yeast two-hybrid assays show the interaction of Borealin10�109 with Survivin and INCENP (via its N-terminal 58 residues Figure 2A birc5-cdca8-4 2 9 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 ecobb Escherichia coli (strain B / BL21) 7 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Gel filtration profiles of Survivin�Borealin10�109 Figure 2C birc5-cdca8-5 2 9 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 ecobb Escherichia coli (strain B / BL21) 7 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) 10 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Gel filtration profiles of Survivin�Borealin10�109 (in red, peak 1) and of Survivin�Borealin10�109�INCENP1�58, (blue, peak 2) formed in the presence of an excess of INCENP1�58 (blue, peak 3). Figure 2C birc5-cdca8-6 6 7 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) 9 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Figure 2D 3.3 s dimer of molecular weight of 53 kDa birc5-cdca8-7 6 7 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) 10 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 ecobb Escherichia coli (strain B / BL21) 9 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction By analytical ultracentrifugation, Survivin�Borealin10�109�INCENP1�58 sedimented at a molecular mass of 35 kDa (Figure 2D), consistent with a complex of 1:1:1 stoichiometry (calculated molecular weight 34.7 kDa) Figure 2D 3.0 s monomer of 35 kDa birc5-cdca8-8 5 9 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 ecobb Escherichia coli (strain B / BL21) 7 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) 10 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 ecobb Escherichia coli (strain B / BL21) physical association physical association The Survivin�Borealin10�109�INCENP1�58 complex was crystallized and its structure determined by single-wavelength anomalous dispersion (SAD). 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. aurkb-cdca8-1 1 10 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 9 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 region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Borealin W70E,F74E mutant binds INCENP and Aurora B but not Survivin. Figure 1F, 5D Mutation of CDCA8 at W70E,F74E has no effect on this interaction but prevents binding of BIRC5 birc5-cdca8-9 1 7 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) 9 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 region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Borealin R35E,L46Y binds Survivin but not INCENP/Aurora B 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 1 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 10 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 association association Figure 6B birc5-cdca8-11 1 10 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 8 unspecified role unspecified role prey prey physical association physical association Figure 6B INCENP F22R,L34R failed to bind Borealin and Survivin but retained binding to Aurora B. Mutation of INCENP F22R,L34R had no effect on thi interaction but prevented binding to BIRC5 and CDCA8.