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/ tsang-2006-1 CP110 cooperates with two calcium-binding proteins to regulate cytokinesis and genome stability. 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 Tsang WY., Spektor A., Luciano DJ., Indjeian VB., Chen Z., Salisbury JL., Sanchez I., Dynlacht BD. A unique UniProtKB entry could not be associated with human alpha-actinin, beta-spectrin, beta-tubulin or DnaJ/Hsp40 homolog since there are several different types of each protein present within the UniProtKB database and the authors have not been specific enough in order for the exact gene names of interactor to be ascertained. brian.dynlacht@med.nyu.edu A human fetal brain cDNA library was used. Mol. Biol. Cell (1059-1524) 2006 tsang-2006-2 CP110 cooperates with two calcium-binding proteins to regulate cytokinesis and genome stability. 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 bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Tsang WY., Spektor A., Luciano DJ., Indjeian VB., Chen Z., Salisbury JL., Sanchez I., Dynlacht BD. A unique UniProtKB entry could not be associated with human alpha-actinin, beta-spectrin, beta-tubulin or DnaJ/Hsp40 homolog since there are several different types of each protein present within the UniProtKB database and the authors have not been specific enough in order for the exact gene names of interactor to be ascertained. brian.dynlacht@med.nyu.edu A human fetal brain cDNA library was used. Mol. Biol. Cell (1059-1524) 2006 tsang-2006-3 CP110 cooperates with two calcium-binding proteins to regulate cytokinesis and genome stability. human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Tsang WY., Spektor A., Luciano DJ., Indjeian VB., Chen Z., Salisbury JL., Sanchez I., Dynlacht BD. A unique UniProtKB entry could not be associated with human alpha-actinin, beta-spectrin, beta-tubulin or DnaJ/Hsp40 homolog since there are several different types of each protein present within the UniProtKB database and the authors have not been specific enough in order for the exact gene names of interactor to be ascertained. brian.dynlacht@med.nyu.edu A human fetal brain cDNA library was used. Mol. Biol. Cell (1059-1524) 2006 tsang-2006-5 CP110 cooperates with two calcium-binding proteins to regulate cytokinesis and genome stability. 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 immunostaining immunostaining Tsang WY., Spektor A., Luciano DJ., Indjeian VB., Chen Z., Salisbury JL., Sanchez I., Dynlacht BD. A unique UniProtKB entry could not be associated with human alpha-actinin, beta-spectrin, beta-tubulin or DnaJ/Hsp40 homolog since there are several different types of each protein present within the UniProtKB database and the authors have not been specific enough in order for the exact gene names of interactor to be ascertained. brian.dynlacht@med.nyu.edu A human fetal brain cDNA library was used. Mol. Biol. Cell (1059-1524) 2006 tsang-2006-6 CP110 cooperates with two calcium-binding proteins to regulate cytokinesis and genome stability. human-t98g Human caucasian glioblastoma line T98G t98g Human Caucasian glioblastoma T98G coip coimmunoprecipitation co-immunoprecipitation CoIp immunoprecipitation Co-IP co-immunoprecipitation CoIp immunoprecipitation Co-IP coip western blot western blot Immuno blot Immuno blot Tsang WY., Spektor A., Luciano DJ., Indjeian VB., Chen Z., Salisbury JL., Sanchez I., Dynlacht BD. To determine whether CP110 and CaM interact before mitosis, the cells were synchronized by serum deprivation and immunoprecipitated CP110 from extracts of cells synchronously traversing the cell cycle. A unique UniProtKB entry could not be associated with human alpha-actinin, beta-spectrin, beta-tubulin or DnaJ/Hsp40 homolog since there are several different types of each protein present within the UniProtKB database and the authors have not been specific enough in order for the exact gene names of interactor to be ascertained. brian.dynlacht@med.nyu.edu A human fetal brain cDNA library was used. Mol. Biol. Cell (1059-1524) 2006 tsang-2006-7 CP110 cooperates with two calcium-binding proteins to regulate cytokinesis and genome stability. in vitro In vitro pull down pull down autoradiography autoradiography Tsang WY., Spektor A., Luciano DJ., Indjeian VB., Chen Z., Salisbury JL., Sanchez I., Dynlacht BD. A unique UniProtKB entry could not be associated with human alpha-actinin, beta-spectrin, beta-tubulin or DnaJ/Hsp40 homolog since there are several different types of each protein present within the UniProtKB database and the authors have not been specific enough in order for the exact gene names of interactor to be ascertained. brian.dynlacht@med.nyu.edu A human fetal brain cDNA library was used. Mol. Biol. Cell (1059-1524) 2006 tsang-2006-9 CP110 cooperates with two calcium-binding proteins to regulate cytokinesis and genome stability. 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. pull down pull down western blot western blot Immuno blot Immuno blot Tsang WY., Spektor A., Luciano DJ., Indjeian VB., Chen Z., Salisbury JL., Sanchez I., Dynlacht BD. A unique UniProtKB entry could not be associated with human alpha-actinin, beta-spectrin, beta-tubulin or DnaJ/Hsp40 homolog since there are several different types of each protein present within the UniProtKB database and the authors have not been specific enough in order for the exact gene names of interactor to be ascertained. brian.dynlacht@med.nyu.edu A human fetal brain cDNA library was used. Mol. Biol. Cell (1059-1524) 2006 cp110_human Centriolar coiled-coil protein of 110 kDa CP110 CCP110 CEP110 KIAA0419 Centrosomal protein of 110 kDa protein protein human Homo sapiens Human MEEYEKFCEKSLARIQEASLSTESFLPAQSESISLIRFHGVAILSPLLNIEKRKEMQQEKQKALDVEARKQVNRKKALLTRVQEILDNVQVRKAPNASDFDQWEMETVYSNSEVRNLNVPATFPNSFPSHTEHSTAAKLDKIAGILPLDNEDQCKTDGIDLARDSEGFNSPKQCDSSNISHVENEAFPKTSSATPQETLISDGPFSVNEQQDLPLLAEVIPDPYVMSLQNLMKKSKEYIEREQSRRSLRGSINRIVNESHLDKEHDAVEVADCVKEKGQLTGKHCVSVIPDKPSLNKSNVLLQGASTQASSMSMPVLASFSKVDIPIRTGHPTVLESNSDFKVIPTFVTENNVIKSLTGSYAKLPSPEPSMSPKMHRRRSRTSSACHILINNPINACELSPKGKEQAMDLIIQDTDENTNVPEIMPKLPTDLAGVCSSKVYVGKNTSEVKEDVVLGKSNQVCQSSGNHLENKVTHGLVTVEGQLTSDERGAHIMNSTCAAMPKLHEPYASSQCIASPNFGTVSGLKPASMLEKNCSLQTELNKSYDVKNPSPLLMQNQNTRQQMDTPMVSCGNEQFLDNSFEKVKRRLDLDIDGLQKENCPYVITSGITEQERQHLPEKRYPKGSGFVNKNKMLGTSSKESEELLKSKMLAFEEMRKRLEEQHAQQLSLLIAEQEREQERLQKEIEEQEKMLKEKKAMTAEASELDINNAVELEWRKISDSSLLETMLSQADSLHTSNSNSSGFTNSAMQYSFVSANEAPFYLWGSSTSGLTKLSVTRPFGRAKTRWSQVFSLEIQAKFNKITAVAKGFLTRRLMQTDKLKQLRQTVKDTMEFIRSFQSEAPLKRGIVSAQDASLQERVLAQLRAALYGIHDIFFVMDAAERMSILHHDREVRKEKMLRQMDKMKSPRVALSAATQKSLDRKKYMKAAEMGMPNKKFLVKQNPSETRVLQPNQGQNAPVHRLLSRQGTPKTSVKGVVQNRQKPSQSRVPNRVPVSGVYAGKIQRKRPNVATI calm_human Calmodulin CALM1 CALM CAM CAM1 CALM2 CAMB CALM3 CAM3 CAMC CAMIII CAM2 CALML2 protein protein human Homo sapiens Human MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK pnma1_human Paraneoplastic antigen Ma1 PNMA1 MA1 Neuron- and testis-specific protein 1 37 kDa neuronal protein protein protein human Homo sapiens Human MAMTLLEDWCRGMDVNSQRALLVWGIPVNCDEAEIEETLQAAMPQVSYRMLGRMFWREENAKAALLELTGAVDYAAIPREMPGKGGVWKVLFKPPTSDAEFLERLHLFLAREGWTVQDVARVLGFQNPTPTPGPEMPAEMLNYILDNVIQPLVESIWYKRLTLFSGRDIPGPGEETFDPWLEHTNEVLEEWQVSDVEKRRRLMESLRGPAADVIRILKSNNPAITTAECLKALEQVFGSVESSRDAQIKFLNTYQNPGEKLSAYVIRLEPLLQKVVEKGAIDKDNVNQARLEQVIAGANHSGAIRRQLWLTGAGEGPAPNLFQLLVQIREEEAKEEEEEAEATLLQLGLEGHF tfp11_human Tuftelin-interacting protein 11 TFIP11 HSPC006 STIP Septin and tuftelin-interacting protein 1 protein protein human Homo sapiens Human MSLSHLYRDGEGRIDDDDDERENFEITDWDLQNEFNPNRQRHWQTKEEATYGVWAERDSDDERPSFGGKRARDYSAPVNFISAGLKKGAAEEAELEDSDDEEKPVKQDDFPKDFGPRKLKTGGNFKPSQKGFAGGTKSFMDFGSWERHTKGIGQKLLQKMGYVPGRGLGKNAQGIINPIEAKQRKGKGAVGAYGSERTTQSMQDFPVVDSEEEAEEEFQKELSQWRKDPSGSKKKPKYSYKTVEELKAKGRISKKLTAPQKELSQVKVIDMTGREQKVYYSYSQISHKHNVPDDGLPLQSQQLPQSGKEAKAPGFALPELEHNLQLLIDLTEQEIIQNDRQLQYERDMVVNLFHELEKMTEVLDHEERVISNLSKVLEMVEECERRMQPDCSNPLTLDECARIFETLQDKYYEEYRMSDRVDLAVAIVYPLMKEYFKEWDPLKDCTYGTEIISKWKSLLENDQLLSHGGQDLSADAFHRLIWEVWMPFVRNIVTQWQPRNCDPMVDFLDSWVHIIPVWILDNILDQLIFPKLQKEVENWNPLTDTVPIHSWIHPWLPLMQARLEPLYSPIRSKLSSALQKWHPSDSSAKLILQPWKDVFTPGSWEAFMVKNIVPKLGMCLGELVINPHQQHMDAFYWVIDWEGMISVSSLVGLLEKHFFPKWLQVLCSWLSNSPNYEEITKWYLGWKSMFSDQVLAHPSVKDKFNEALDIMNRAVSSNVGAYMQPGARENIAYLTHTERRKDFQYEAMQERREAENMAQRGIGVAASSVPMNFKDLIETKAEEHNIVFMPVIGKRHEGKQLYTFGRIVIYIDRGVVFVQGEKTWVPTSLQSLIDMAK myp0_human Myelin protein P0 Myelin protein zero Myelin peripheral protein MPZ protein protein human Homo sapiens Human MAPGAPSSSPSPILAVLLFSSLVLSPAQAIVVYTDREVHGAVGSRVTLHCSFWSSEWVSDDISFTWRYQPEGGRDAISIFHYAKGQPYIDEVGTFKERIQWVGDPRWKDGSIVIHNLDYSDNGTFTCDVKNPPDIVGKTSQVTLYVFEKVPTRYGVVLGAVIGGVLGVVLLLLLLFYVVRYCWLRRQAALQRRLSAMEKGKLHKPGKDASKRGRQTPVLYAMLDHSRSTKAVSEKKAKGLGESRKDKK cetn2_human Centrin-2 CETN2 CALT CEN2 Caltractin isoform 1 protein protein human Homo sapiens Human MASNFKKANMASSSQRKRMSPKPELTEEQKQEIREAFDLFDADGTGTIDVKELKVAMRALGFEPKKEEIKKMISEIDKEGTGKMNFGDFLTVMTQKMSEKDTKEEILKAFKLFDDDETGKISFKNLKRVAKELGENLTDEELQEMIDEADRDGDGEVSEQEFLRIMKKTSLY cep110-cam Y2H showing interaction between Cep110 and Calmodulin 695539 N/A 695546 unspecified role unspecified role bait bait N/A 695547 unspecified role unspecified role prey prey physical association physical association false false false Figure 1A. cep110-pnma Y2H showing interaction between Cep110 and PNMA1 (paraneoplastic antigen MA1) 695539 N/A 695546 unspecified role unspecified role bait bait N/A 695548 unspecified role unspecified role prey prey physical association physical association false false false Figure 1A. cep110-tfip Y2H showing interaction between Cep110 and tfip (Tuftelin-interacting protein) 695539 N/A 695549 unspecified role unspecified role prey prey N/A 695546 unspecified role unspecified role bait bait physical association physical association false false false Figure 1A. cep110-mpz Y2H showing interaction between Cep110 and mpz (peripheral myelin protein) 695539 N/A 695550 unspecified role unspecified role prey prey N/A 695546 unspecified role unspecified role bait bait physical association physical association false false false Figure 1A. cep110-cam-1 CoIP showing interaction between endogenous Cep110 and Calmodulin 695540 N/A 695547 unspecified role unspecified role prey prey N/A 695546 unspecified role unspecified role bait bait physical association physical association false false false Figure 1B, 1D, 1E. Endogenous CP110 binds specifically to CaM in the presence or absence of Ca2+ suggesting that CP110 may contain both Ca2+-dependent and Ca2+-independent CaM-binding domains. cep110-centrin CoIP showing interaction between endogenous Cep110 and Centrin 695540 N/A 695546 unspecified role unspecified role prey prey N/A 695551 unspecified role unspecified role bait bait physical association physical association false false false Figure 5C and 5E. Immunoprecipitation experiments indicated that CP110 associated with centrin in a subset of the high-molecular-weight fractions in which the proteins coeluted (fractions 16-18). cep110-centrin-2 CoIP showing interaction of mutant CEP110 with endogenous Centrin 695540 N/A 695551 unspecified role unspecified role bait bait N/A 695546 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged western blot western blot Immuno blot Immuno blot undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false 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 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 Figure 9C. Both the full-length CEP110 and mutant CEP110 proteins (missing regions 67-82, 781-820 and 914-933) interacted with endogenous Centrin as detected using anti-centrin antibody. cep110-cam-8 CoIP showing interaction of mutant and full-length CEP110 with endogenous Calmodulin 695540 N/A 695547 unspecified role unspecified role prey prey 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. N/A 695546 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged anti tag western anti tag western blot anti tag western undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region required to bind necessary binding region required to bind deletion analysis deletion 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 in vitro In vitro physical association physical association false false false Figure 9D. Only the full-length CEP110 interacted with endogenous Calmodulin as detected using anti-FLAG antibody suggesting that the regions that were deleted were important for this interaction. The deletion mutant of CEP110 did display some residual binding. cep110-cam-2 CoIP showing interaction between endogenous Cep110 and Calmodulin 695541 N/A 695546 unspecified role unspecified role prey prey N/A 695547 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 physical association physical association false false false Figure 1C. cep110-cam-4 Colocalization of endogenous Cep110 and Calmodulin during mitosis. 695542 N/A 695546 unspecified role unspecified role neutral component neutral component N/A 695547 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Figure 2A. CP110 and a portion of CaM colocalized with substantial overlapping during the metaphase, anaphase and telophase stages of mitosis. cep110-cam-5 CoIP showing interaction of endogenous Cep110 and Calmodulin throughout the cell cycle. 695543 N/A 695547 unspecified role unspecified role prey prey N/A 695546 unspecified role unspecified role bait bait physical association physical association false false false Figure 2B. CP110 and CaM associate at the centrosome throughout the cell cycle, although the interaction is most easily visualized at the cellular level during mitosis. cep110-cam-7 In vitro binding assay showing interaction regions of Cep110 for binding with Calmodulin. 695544 N/A 695546 unspecified role unspecified role prey prey n-terminus sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false c-terminus 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- and c- termini required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region required to bind necessary binding region required to bind deletion analysis deletion 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 region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 autoradiography autoradiography undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 695547 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 false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 3A, 3B, 3C, 4A, 4B. This experiment showed that the cep110 protein contains independent n-terminal and c-terminal CaM-binding domains (3 CaM-binding domains were predicted by the Calmodulin Target Database at residues 64-82, 781-821 and 909-924) that are Ca2+-dependent and are required for the interaction. Additional Ca2+-independent CaM binding sites, not revealed by the database, may be present at the carboxy terminus as shown when weak residual interaction occurs between CaM and full-length CP110 or carboxy-terminal fragments when calcium is absent and even when the putative CaM-binding domains were all deleted together. cep110-centrin-1 Pull-down showing interaction between endogenous Cep110 and Centrin 695545 N/A 695551 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 false ecoli Escherichia coli (strain K12) N/A 695546 unspecified role unspecified role prey prey 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. physical association physical association false false false Figure 5D.