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/ Interaction of the transforming acidic coiled-coil 1 (TACC1) protein with ch-TOG and GAS41/NuBI1 suggests multiple TACC1-containing protein complexes in human cells. Interaction of the transforming acidic coiled-coil 1 (TACC1) protein with ch-TOG and GAS41/NuBI1 suggests multiple TACC1-containing protein complexes in human cells. Biochem. J. (0264-6021) 2002 Lauffart B., Howell SJ., Tasch JE., Cowell JK., Still IH. Cancer - Interactions investigated in the context of cancer Full-length GAS41 fused to GFP was expressed in stably transfected cells and indirect immunofluoresence with the TACC1 antibody and a rhodamine-labelled secondary antibody was used. Brenda.Laufert@roswellpark.org 01-DEC-2006: Contacted by JYOTI. A human mammary epithelial cDNA library from ClonTech Laboratories (Palo Alto, CA, U.S.A.) was used. human-mda_mb_468 Homo sapiens MDA-MB-468 mda_mb_468 MDA-MB-468 breast adenocarcinoma Species: human, Black; Tissue: breast; Tumor: adenocarcinoma; Derived from: pleural effusion fluorescence imaging fluorescence microscopy fluorescence imaging predetermined predetermined participant predetermined Lauffart B., Howell SJ., Tasch JE., Cowell JK., Still IH. Cancer - Interactions investigated in the context of cancer Full-length GAS41 fused to GFP was expressed in stably transfected cells and indirect immunofluoresence with the TACC1 antibody and a rhodamine-labelled secondary antibody was used. Brenda.Laufert@roswellpark.org Accepted 2006-NOV-9 by JYOTI. 01-DEC-2006: Contacted by JYOTI. A human mammary epithelial cDNA library from ClonTech Laboratories (Palo Alto, CA, U.S.A.) was used. Biochem. J. (0264-6021) 2002 Interaction of the transforming acidic coiled-coil 1 (TACC1) protein with ch-TOG and GAS41/NuBI1 suggests multiple TACC1-containing protein complexes in human cells. Interaction of the transforming acidic coiled-coil 1 (TACC1) protein with ch-TOG and GAS41/NuBI1 suggests multiple TACC1-containing protein complexes in human cells. Biochem. J. (0264-6021) 2002 Lauffart B., Howell SJ., Tasch JE., Cowell JK., Still IH. Cancer - Interactions investigated in the context of cancer Full-length GAS41 fused to GFP was expressed in stably transfected cells and indirect immunofluoresence with the TACC1 antibody and a rhodamine-labelled secondary antibody was used. Brenda.Laufert@roswellpark.org 01-DEC-2006: Contacted by JYOTI. A human mammary epithelial cDNA library from ClonTech Laboratories (Palo Alto, CA, U.S.A.) was used. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Lauffart B., Howell SJ., Tasch JE., Cowell JK., Still IH. Cancer - Interactions investigated in the context of cancer Full-length GAS41 fused to GFP was expressed in stably transfected cells and indirect immunofluoresence with the TACC1 antibody and a rhodamine-labelled secondary antibody was used. Brenda.Laufert@roswellpark.org Accepted 2006-NOV-9 by JYOTI. 01-DEC-2006: Contacted by JYOTI. A human mammary epithelial cDNA library from ClonTech Laboratories (Palo Alto, CA, U.S.A.) was used. Biochem. J. (0264-6021) 2002 Interaction of the transforming acidic coiled-coil 1 (TACC1) protein with ch-TOG and GAS41/NuBI1 suggests multiple TACC1-containing protein complexes in human cells. Interaction of the transforming acidic coiled-coil 1 (TACC1) protein with ch-TOG and GAS41/NuBI1 suggests multiple TACC1-containing protein complexes in human cells. Biochem. J. (0264-6021) 2002 Lauffart B., Howell SJ., Tasch JE., Cowell JK., Still IH. Cancer - Interactions investigated in the context of cancer Full-length GAS41 fused to GFP was expressed in stably transfected cells and indirect immunofluoresence with the TACC1 antibody and a rhodamine-labelled secondary antibody was used. Brenda.Laufert@roswellpark.org 01-DEC-2006: Contacted by JYOTI. A human mammary epithelial cDNA library from ClonTech Laboratories (Palo Alto, CA, U.S.A.) was used. 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 Lauffart B., Howell SJ., Tasch JE., Cowell JK., Still IH. Cancer - Interactions investigated in the context of cancer Full-length GAS41 fused to GFP was expressed in stably transfected cells and indirect immunofluoresence with the TACC1 antibody and a rhodamine-labelled secondary antibody was used. Brenda.Laufert@roswellpark.org Accepted 2006-NOV-9 by JYOTI. 01-DEC-2006: Contacted by JYOTI. A human mammary epithelial cDNA library from ClonTech Laboratories (Palo Alto, CA, U.S.A.) was used. Biochem. J. (0264-6021) 2002 Interaction of the transforming acidic coiled-coil 1 (TACC1) protein with ch-TOG and GAS41/NuBI1 suggests multiple TACC1-containing protein complexes in human cells. Interaction of the transforming acidic coiled-coil 1 (TACC1) protein with ch-TOG and GAS41/NuBI1 suggests multiple TACC1-containing protein complexes in human cells. Biochem. J. (0264-6021) 2002 Lauffart B., Howell SJ., Tasch JE., Cowell JK., Still IH. Cancer - Interactions investigated in the context of cancer Full-length GAS41 fused to GFP was expressed in stably transfected cells and indirect immunofluoresence with the TACC1 antibody and a rhodamine-labelled secondary antibody was used. Brenda.Laufert@roswellpark.org 01-DEC-2006: Contacted by JYOTI. A human mammary epithelial cDNA library from ClonTech Laboratories (Palo Alto, CA, U.S.A.) was used. 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 Lauffart B., Howell SJ., Tasch JE., Cowell JK., Still IH. Cancer - Interactions investigated in the context of cancer Full-length GAS41 fused to GFP was expressed in stably transfected cells and indirect immunofluoresence with the TACC1 antibody and a rhodamine-labelled secondary antibody was used. Brenda.Laufert@roswellpark.org Accepted 2006-NOV-9 by JYOTI. 01-DEC-2006: Contacted by JYOTI. A human mammary epithelial cDNA library from ClonTech Laboratories (Palo Alto, CA, U.S.A.) was used. Biochem. J. (0264-6021) 2002 Interaction of the transforming acidic coiled-coil 1 (TACC1) protein with ch-TOG and GAS41/NuBI1 suggests multiple TACC1-containing protein complexes in human cells. Interaction of the transforming acidic coiled-coil 1 (TACC1) protein with ch-TOG and GAS41/NuBI1 suggests multiple TACC1-containing protein complexes in human cells. Biochem. J. (0264-6021) 2002 Lauffart B., Howell SJ., Tasch JE., Cowell JK., Still IH. Cancer - Interactions investigated in the context of cancer Full-length GAS41 fused to GFP was expressed in stably transfected cells and indirect immunofluoresence with the TACC1 antibody and a rhodamine-labelled secondary antibody was used. Brenda.Laufert@roswellpark.org 01-DEC-2006: Contacted by JYOTI. A human mammary epithelial cDNA library from ClonTech Laboratories (Palo Alto, CA, U.S.A.) was used. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. fluorescence imaging fluorescence microscopy fluorescence imaging predetermined predetermined participant predetermined Lauffart B., Howell SJ., Tasch JE., Cowell JK., Still IH. Cancer - Interactions investigated in the context of cancer Full-length GAS41 fused to GFP was expressed in stably transfected cells and indirect immunofluoresence with the TACC1 antibody and a rhodamine-labelled secondary antibody was used. Brenda.Laufert@roswellpark.org Accepted 2006-NOV-9 by JYOTI. 01-DEC-2006: Contacted by JYOTI. A human mammary epithelial cDNA library from ClonTech Laboratories (Palo Alto, CA, U.S.A.) was used. Biochem. J. (0264-6021) 2002 yets4_human YEATS domain-containing protein 4 YEATS4 Glioma-amplified sequence 41 NuMA-binding protein 1 GAS41 protein protein human Homo sapiens Human MFKRMAEFGPDSGGRVKGVTIVKPIVYGNVARYFGKKREEDGHTHQWTVYVKPYRNEDMSAYVKKIQFKLHESYGNPLRVVTKPPYEITETGWGEFEIIIKIFFIDPNERPVTLYHLLKLFQSDTNAMLGKKTVVSEFYDEMIFQDPTAMMQQLLTTSRQLTLGAYKHETEFAELEVKTREKLEAAKKKTSFEIAELKERLKASRETINCLKNEIRKLEEDDQAKDI 34C4692B227A7B85 tacc1_human Transforming acidic coiled-coil-containing protein 1 Taxin-1 Gastric cancer antigen Ga55 TACC1 KIAA1103 protein protein human Homo sapiens Human MAFSPWQILSPVQWAKWTWSAVRGGAAGEDEAGGPEGDPEEEDSQAETKSLSFSSDSEGNFETPEAETPIRSPFKESCDPSLGLAGPGAKSQESQEADEQLVAEVVEKCSSKTCSKPSENEVPQQAIDSHSVKNFREEPEHDFSKISIVRPFSIETKDSTDISAVLGTKAAHGCVTAVSGKALPSSPPDALQDEAMTEGSMGVTLEASAEADLKAGNSCPELVPSRRSKLRKPKPVPLRKKAIGGEFSDTNAAVEGTPLPKASYHFSPEELDENTSPLLGDARFQKSPPDLKETPGTLSSDTNDSGVELGEESRSSPLKLEFDFTEDTGNIEARKALPRKLGRKLGSTLTPKIQKDGISKSAGLEQPTDPVARDGPLSQTSSKPDPSQWESPSFNPFGSHSVLQNSPPLSSEGSYHFDPDNFDESMDPFKPTTTLTSSDFCSPTGNHVNEILESPKKAKSRLITSGCKVKKHETQSLALDACSRDEGAVISQISDISNRDGHATDEEKLASTSCGQKSAGAEVKGEPEEDLEYFECSNVPVSTINHAFSSSEAGIEKETCQKMEEDGSTVLGLLESSAEKAPVSVSCGGESPLDGICLSESDKTAVLTLIREEIITKEIEANEWKKKYEETRQEVLEMRKIVAEYEKTIAQMIEDEQRTSMTSQKSFQQLTMEKEQALADLNSVERSLSDLFRRYENLKGVLEGFKKNEEALKKCAQDYLARVKQEEQRYQALKIHAEEKLDKANEEIAQVRTKAKAESAALHAGLRKEQMKVESLERALQQKNQEIEELTKICDELIAKLGKTD 3A261EF58C165107 ckap5_human Cytoskeleton-associated protein 5 CKAP5 KIAA0097 Colonic and hepatic tumor overexpressed gene protein protein protein human Homo sapiens Human MGDDSEWLKLPVDQKCEHKLWKARLSGYEEALKIFQKIKDEKSPEWSKFLGLIKKFVTDSNAVVQLKGLEAALVYVENAHVAGKTTGEVVSGVVSKVFNQPKAKAKELGIEICLMYIEIEKGEAVQEELLKGLDNKNPKIIVACIETLRKALSEFGSKIILLKPIIKVLPKLFESREKAVRDEAKLIAVEIYRWIRDALRPPLQNINSVQLKELEEEWVKLPTSAPRPTRFLRSQQELEAKLEQQQSAGGDAEGGGDDGDEVPQIDAYELLEAVEILSKLPKDFYDKIEAKKWQERKEALESVEVLIKNPKLEAGDYADLVKALKKVVGKDTNVMLVALAAKCLTGLAVGLRKKFGQYAGHVVPTILEKFKEKKPQVVQALQEAIDAIFLTTTLQNISEDVLAVMDNKNPTIKQQTSLFIARSFRHCTASTLPKSLLKPFCAALLKHINDSAPEVRDAAFEALGTALKVVGEKAVNPFLADVDKLKLDKIKECSEKVELIHGKKAGLAADKKEFKPLPGRTAASGAAGDKDTKDISAPKPGPLKKAPAAKAGGPPKKGKPAAPGGAGNTGTKNKKGLETKEIVEPELSIEVCEEKASAVLPPTCIQLLDSSNWKERLACMEEFQKAVELMDRTEMPCQALVRMLAKKPGWKETNFQVMQMKLHIVALIAQKGNFSKTSAQVVLDGLVDKIGDVKCGNNAKEAMTAIAEACMLPWTAEQVVSMAFSQKNPKNQSETLNWLSNAIKEFGFSGLNVKAFISNVKTALAATNPAVRTAAITLLGVMYLYVGPSLRMFFEDEKPALLSQIDAEFEKMQGQSPPAPTRGISKHSTSGTDEGEDGDEPDDGSNDVVDLLPRTEISDKITSELVSKIGDKNWKIRKEGLDEVAGIINDAKFIQPNIGELPTALKGRLNDSNKILVQQTLNILQQLAVAMGPNIKQHVKNLGIPIITVLGDSKNNVRAAALATVNAWAEQTGMKEWLEGEDLSEELKKENPFLRQELLGWLAEKLPTLRSTPTDLILCVPHLYSCLEDRNGDVRKKAQDALPFFMMHLGYEKMAKATGKLKPTSKDQVLAMLEKAKVNMPAKPAPPTKATSKPMGGSAPAKFQPASAPAEDCISSSTEPKPDPKKAKAPGLSSKAKSAQGKKMPSKTSLKEDEDKSGPIFIVVPNGKEQRMKDEKGLKVLKWNFTTPRDEYIEQLKTQMSSCVAKWLQDEMFHSDFQHHNKALAVMVDHLESEKEGVIGCLDLILKWLTLRFFDTNTSVLMKALEYLKLLFTLLSEEEYHLTENEASSFIPYLVVKVGEPKDVIRKDVRAILNRMCLVYPASKMFPFIMEGTKSKNSKQRAECLEELGCLVESYGMNVCQPTPGKALKEIAVHIGDRDNAVRNAALNTIVTVYNVHGDQVFKLIGNLSEKDMSMLEERIKRSAKRPSAAPIKQVEEKPQRAQNISSNANMLRKGPAEDMSSKLNQARSMSGHPEAAQMVRREFQLDLDEIENDNGTVRCEMPELVQHKLDDIFEPVLIPEPKIRAVSPHFDDMHSNTASTINFIISQVASGDINTSIQALTQIDEVLRQEDKAEAMSGHIDQFLIATFMQLRLIYNTHMADEKLEKDEIIKLYSCIIGNMISLFQIESLAREASTGVLKDLMHGLITLMLDSRIEDLEEGQQVIRSVNLLVVKVLEKSDQTNILSALLVLLQDSLLATASSPKFSELVMKCLWRMVRLLPDTINSINLDRILLDIHIFMKVFPKEKLKQCKSEFPIRTLKTLLHTLCKLKGPKILDHLTMIDNKNESELEAHLCRMMKHSMDQTGSKSDKETEKGASRIDEKSSKAKVNDFLAEIFKKIGSKENTKEGLAELYEYKKKYSDADIEPFLKNSSQFFQSYVERGLRVIEMEREGKGRISTSTGISPQMEVTCVPTPTSTVSSIGNTNGEEVGPSVYLERLKILRQRCGLDNTKQDDRPPLTSLLSKPAVPTVASSTDMLHSKLSQLRESREQHQHSDLDSNQTHSSGTVTSSSSTANIDDLKKRLERIKSSRK B2BBFB1CF2ED688B tacc1-ckap5 4 8 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind range range range range 7 unspecified role unspecified role bait bait physical association physical association Subsequent isolation and sequence analysis revealed that four of these clones corresponded to the C-terminal 529 amino acids of a previously identi�ed TACC binding protein, the human ch-TOG protein 1 A The authors state TACC1 interacted with the protein corresponding to the C-terminal 529 amino acids of ch-TOG and that this starts at residue 1443. However, this would make the protein 1972 residues in length whereas the UniprotKB entry is 2032 aa in length. The ch-TOG interactor has a feature describing the C-terminal 529 amino acids (which does contain the KXGS motif described) but starts at residue 1704. The sequence comparison of SDP repeats in Figure 2 has slightly different residue positions for TACC1 than the UniprotKb entry for TACC1. tacc1-yeats4 4 6 unspecified role unspecified role prey prey c-terminal TACIP 38 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role bait bait physical association physical association 1 B Subsequent isolation and sequence analysis revealed that four of these clones corresponded to the C-terminal 529 amino acids of a previously identi�ed TACC binding protein, the human ch-TOG protein (Figure 1A), and that �ve corresponded to the C-terminal region of the putative oncogenic transcription factor GAS41}NuBI1 The sequence comparison of SDP repeats in Figure 2 has slightly different residue positions for TACC1 than the UniprotKb entry for TACC1. tacc1-yeats4-1 3 6 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role bait bait physical association physical association 1 B To determine whether the TACC1 binding site on GAS41}NuBI1 overlapped with that for NuMA, we generated an additional smaller construct containing the C-terminal amino acids 168�227. This construct activated the His3 and LacZ reporter genes in the diploid strain only in the presence of the pASTACC1 construct The sequence comparison of SDP repeats in Figure 2 has slightly different residue positions for TACC1 than the UniprotKb entry for TACC1. tacc1-yeats4-2 3 7 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 6 unspecified role unspecified role prey prey c-terminal TACIP 38 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 2 A a series of smaller TACC constructs were constructed and expressed as GAL4 DNA-binding domain constructs in yeast strain Y187. TACIP38 (in yeast strain CG1945) was mated to each construct and assayed for speci�c interaction with each region of TACC1. Interaction was only obtained with constructs containing amino acids 206�427 The sequence comparison of SDP repeats in Figure 2 has slightly different residue positions for TACC1 than the UniprotKb entry for TACC1. yeats4-tacc1 2 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait c-terminus GFP tag fluorescent prot tag fluorescent protein tag fluorescent prot tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association 3 TACC1 speci�cally co-immunoprecipitated with the EGGAS41 fusion protein, using an antibody raised against the GFP moiety The sequence comparison of SDP repeats in Figure 2 has slightly different residue positions for TACC1 than the UniprotKb entry for TACC1. tacc1-yeats4-3 5 6 unspecified role unspecified role neutral component neutral component c-terminus GFP tag fluorescent prot tag fluorescent protein tag fluorescent prot tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 7 unspecified role unspecified role neutral component neutral component colocalization colocalization 4 Indirect immuno�uorescence microscopy using the TACC1 antibody revealed a similar staining pattern to the EGGAS41 fusion protein, This experiment suggests that the primary site of interaction between GAS41 and TACC1 is the nuceus. The sequence comparison of SDP repeats in Figure 2 has slightly different residue positions for TACC1 than the UniprotKb entry for TACC1. tacc1-yeats4-4 1 6 unspecified role unspecified role neutral component neutral component c-terminus GFP tag fluorescent prot tag fluorescent protein tag fluorescent prot tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 7 unspecified role unspecified role neutral component neutral component colocalization colocalization 4 Indirect immuno�uorescence microscopy using the TACC1 antibody revealed a similar staining pattern to the EGGAS41 fusion protein, This experiment suggests that the primary site of interaction between GAS41 and TACC1 is the nuceus. The sequence comparison of SDP repeats in Figure 2 has slightly different residue positions for TACC1 than the UniprotKb entry for TACC1.