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/ CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore. CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore. Cell (0092-8674) 2008 imex curation Hori T., Amano M., Suzuki A., Backer CB., Welburn JP., Dong Y., McEwen BF., Shang WH., Suzuki E., Okawa K., Cheeseman IM., Fukagawa T. I have not captured the information in Fig. 3 as H3 cannot be ascribed to a single gene product and the interaction in Fig 3G is not seen in in 3D when DNA has been digested suggesting proteins are both separately interacting with chromatin, not forming a complex. tfukagaw@lab.nig.ac.jp 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 9 chick-dt40 Gallus gallus bursa lymphoma dt40 chicken bursa lymphoma http://www.lgcpromochem-atcc.com/common/catalog/numSearch/numResults.cfm chicken bursa lymphoma fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Hori T., Amano M., Suzuki A., Backer CB., Welburn JP., Dong Y., McEwen BF., Shang WH., Suzuki E., Okawa K., Cheeseman IM., Fukagawa T. I have not captured the information in Fig. 3 as H3 cannot be ascribed to a single gene product and the interaction in Fig 3G is not seen in in 3D when DNA has been digested suggesting proteins are both separately interacting with chromatin, not forming a complex. tfukagaw@lab.nig.ac.jp Accepted 2009-FEB-14 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 9 imex curation CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore. CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore. Cell (0092-8674) 2008 imex curation Hori T., Amano M., Suzuki A., Backer CB., Welburn JP., Dong Y., McEwen BF., Shang WH., Suzuki E., Okawa K., Cheeseman IM., Fukagawa T. I have not captured the information in Fig. 3 as H3 cannot be ascribed to a single gene product and the interaction in Fig 3G is not seen in in 3D when DNA has been digested suggesting proteins are both separately interacting with chromatin, not forming a complex. tfukagaw@lab.nig.ac.jp 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 9 chick-dt40 Gallus gallus bursa lymphoma dt40 chicken bursa lymphoma http://www.lgcpromochem-atcc.com/common/catalog/numSearch/numResults.cfm chicken bursa lymphoma molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Hori T., Amano M., Suzuki A., Backer CB., Welburn JP., Dong Y., McEwen BF., Shang WH., Suzuki E., Okawa K., Cheeseman IM., Fukagawa T. I have not captured the information in Fig. 3 as H3 cannot be ascribed to a single gene product and the interaction in Fig 3G is not seen in in 3D when DNA has been digested suggesting proteins are both separately interacting with chromatin, not forming a complex. tfukagaw@lab.nig.ac.jp Accepted 2009-FEB-14 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 9 imex curation CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore. CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore. Cell (0092-8674) 2008 imex curation Hori T., Amano M., Suzuki A., Backer CB., Welburn JP., Dong Y., McEwen BF., Shang WH., Suzuki E., Okawa K., Cheeseman IM., Fukagawa T. I have not captured the information in Fig. 3 as H3 cannot be ascribed to a single gene product and the interaction in Fig 3G is not seen in in 3D when DNA has been digested suggesting proteins are both separately interacting with chromatin, not forming a complex. tfukagaw@lab.nig.ac.jp 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 9 chick-dt40 Gallus gallus bursa lymphoma dt40 chicken bursa lymphoma http://www.lgcpromochem-atcc.com/common/catalog/numSearch/numResults.cfm chicken bursa lymphoma anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Hori T., Amano M., Suzuki A., Backer CB., Welburn JP., Dong Y., McEwen BF., Shang WH., Suzuki E., Okawa K., Cheeseman IM., Fukagawa T. I have not captured the information in Fig. 3 as H3 cannot be ascribed to a single gene product and the interaction in Fig 3G is not seen in in 3D when DNA has been digested suggesting proteins are both separately interacting with chromatin, not forming a complex. tfukagaw@lab.nig.ac.jp Accepted 2009-FEB-14 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 9 imex curation CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore. CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore. Cell (0092-8674) 2008 imex curation Hori T., Amano M., Suzuki A., Backer CB., Welburn JP., Dong Y., McEwen BF., Shang WH., Suzuki E., Okawa K., Cheeseman IM., Fukagawa T. I have not captured the information in Fig. 3 as H3 cannot be ascribed to a single gene product and the interaction in Fig 3G is not seen in in 3D when DNA has been digested suggesting proteins are both separately interacting with chromatin, not forming a complex. tfukagaw@lab.nig.ac.jp 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 9 chick-dt40 Gallus gallus bursa lymphoma dt40 chicken bursa lymphoma http://www.lgcpromochem-atcc.com/common/catalog/numSearch/numResults.cfm chicken bursa lymphoma anti tag coip anti tag coimmunoprecipitation anti tag coip ms participant Identification by mass spectrometry ms participant Hori T., Amano M., Suzuki A., Backer CB., Welburn JP., Dong Y., McEwen BF., Shang WH., Suzuki E., Okawa K., Cheeseman IM., Fukagawa T. I have not captured the information in Fig. 3 as H3 cannot be ascribed to a single gene product and the interaction in Fig 3G is not seen in in 3D when DNA has been digested suggesting proteins are both separately interacting with chromatin, not forming a complex. Source of bait proteins supplied by the author tfukagaw@lab.nig.ac.jp Accepted 2009-FEB-14 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 9 imex curation cenpm_chick Centromere protein M CENPM protein protein chick Gallus gallus (Chicken) Chicken MAVLRPFDKLPELNSATILLVGAEGGPQQQLAEAMLREKKDFNINIHLASSLPLPSERDHLRPRIDLIAFVIDIKSKYSLKNVQASLAYVDVRFFLGKVCFLVTGVGRANNCSVEMNAIWKLGEDYCSPVIFCELELEGIRVATAQRLLRMLQICAGRVPGVSALYFSLLMRNSAGD 577DFD82451816C1 o57392_chick centoromere protein C CENP-C protein protein chick Gallus gallus (Chicken) Chicken MAERLDHLKKYYRARFCGQGGKKINIQPGQNVLKYIQDCFECDDTDVTINSPNATSSSTPLVRNQERTSTQSPRPDAGLTIVVGNRTCHSTVSLPATPAETVRSKGEPCVFCPEPGRNSRIADSIKTDSPVHRGETQVLTFKDVEGLCESPAKLSDPEDDPGVVGSPLLVEEAKTPVQTRTVSIAPSSPSPVKDHNVEIESDCEFLIDESGCLSFKSWFSIPRKNRKSQKDGSAAPVLKSQSSDKEKTADKKGKNMNVQAEVHSEQKTKNVRVQRDLKRTSESHPVSSDSEGKVLKSKMQSSTHAGKSKKDALKGSSKHRKKMSYESEAEQLMLSGLDSETSDGEQKIRRVMPREDLAVPSAEHQQEQMLSSKKNLTPCKYQQSASKAYQRSVHKKQTKQTLANDTVLKSQRKKLKKSGKRNGNKNPEPRRWKSSDSEPGEEGLEREPLESNEMFTSPPHQELQTSVLQKLAKSEKAEKVLHALESVTDVNNKTPLKAVELLQHLVDSMKNSEKKRSSVTSSGKMLKKVHHRAHKRACSITRDSEPQNTTDSDSSSAPEVARKKRRQSAVRASKKPKSNRHQRLQHSPAAEKAMGPESGLVLEHSEKYTSPSKHCEENNESSDNSEDLHCQIKNLLSDEIGRQKIVLPSNTPNVRRTKRIRLKPLEYWRGERVTYTLKPSGRLLISGIAGAEAEPHMKNKLKSRHKKKRAKTRSEVVKCLDIPLADASKPTSIVDPVTNQEVLLECINSGSSHSCFFEDESIKVYKSLNTSDFAAGKLILKPLKEKGHQFVHMDTIAFYVIRGQIITTLHKTSYYLTSGDYFYVPAGNGYNIRNLLNEESVLHFTQLKNDRAPVGAELCSVTNENWSLEGKS D345F53B628C63F6 cenpt_chick Centromere protein T CENPT protein protein chick Gallus gallus (Chicken) Chicken MDGRVGLRASRRAAPTPRVAVRSSPRHRARVREQEPVVSAMSGSGLGKENKAGSSTPLRHRPNAFSELDNATPRVMLRKIIQNQPQVSPLALQTVQLEETEDARPEPPSQRTSSTVELQLPDLVPEDASVTTFRMTRKRKKLSISEFERAADKRLPQNQAHSTLDSTLVRSLRMSVGSVMAPDTVEKRGLLRRPQNHKAIDIAAFEGGVEQNMLQIKAQDYLVDLQTSSMTGTTTIRTDAEVVLNNTELFVEPQLGEQNLLAVEPQLSDSKTSAQRSNTSYPAHEKARLEGLVSRVSTDERRTLRFSEKDLITDHEHVDGITQKTPAKQGEEEQDHSQQNDPMEQFSESEEMAGTTEHHADAEYSEHSEKKLSRKAVSQLTAAQDAGVEMEMTPSEGGVAEGTEHQDSPKAELQMAGSPGGHSPASYSLEKPGAKPLKEAVEQTGEIERGTITGVLDAAEEEATDDESDKEDHESEEISMKTPMFVHAAAYRPQPVLSPPHPVKSASPELPPQPVRAKPVPKSSGAAQRKTREPEIASSLIKQIFSHYVKTPVTRDAYKIVEKCSERYFKQISSDLEAYSQHAGRKTVEMADVELLMRRQGLVTDKMPLHVLVERHLPLEYRKLLIPIAVSGNKVIPCK 9D314920A56ADC11 cenpa_chick Histone H3-like centromeric protein A CENPA Centromere protein A protein protein chick Gallus gallus (Chicken) Chicken MPRPKPRSPRRRGRPPPAAPPPPPARPRARRYRPGQRALREIRRYQSSTALLLRRQPFARVVREICLLFTRGVDYRWQAMALLALQEAAEAFLVHLLEDAYLCSLHARRVTLYPKDLQLARRLRGLQGEGF 4D0F421A4CEAD0EF cenph_chick Centromere protein H CENPH protein protein chick Gallus gallus (Chicken) Chicken MAGRLSESVGSGPGAEAETAADPDAKRDVLEHLCARTHLKQLVMEFDTACPPDEGCNSGAEVNFIESAKETLEEVGKVKSAFESKALVIKRIQLMDALRKRVKENDGCARLIVETMRDIIKLNWEIIQAHQQARVIRENLNDIRRKRYFLKQAEGEKALRIFTTVRKKKEVVRMKIAEKLKFIHRNVQYERKVTTLVQNILQNIIVGCQINWAKDPSLRAIILQLEKDISIQNLL 26808B7C1C8A4AC3 cenpi_chick Centromere protein I CENPI protein protein chick Gallus gallus (Chicken) Chicken MQRRQSSKHSKRPLQVHHSNQTDLSAWRKGGTVDTEKSAQNRQSLSDQKNDNEQDSLEQALSYFEKIQDRVSLKKSEVLQKHLSTMESIALKRGLPPEGFDVLLDVALSGKLADTVNTRLLKSLIPASAIPESSIVSSVSWFCVSKCSSNIQLLFLRWLITMFDFIDHKEQVHALYGIFFSFLNDEKLCPYICHVLYLLTRKENVKPFRVRRLLDLQSKMGMQPHLQALLSLYKLFCPELVSITLPQKMKTYFKNADGPWKAAINAVRQRNQANSTVPQPLLLGTAQPHSRKRKWNTQLIVPASSANAQNLVVGGKMSRADSYSANESFPVEQLRTFPQLLQNIHRLEFPSQMGSVLTNPLLLHYMNCSKDESVYLRLYYWMGQTLQEECTWCVVDNNQYEEEFRGFLETVYKAECFLQEGFPSCEEFLYRSLPLWDGVSCRSQILQLVSWIPLSTFSEMKSQLCDPLAQLFFTSSLYFKCSVLESLKELLQNWLNWHVVQLDSESDSQFSSLNTTLSGLVNGVAELINFVGRISTAALHLEKSHTFLLYFILDFYETVCDIYLKYKLPLLIMPPAGVFYPALLSMDSVNLNQLCYIMYRYRTNLIAAKENEMSKKKIQQFKFSSQTYQEYNQYIIAMVGCLWTSSAFQKDNHPEGIRLDDELLKKTGVREYKNSFNIVYHPALMCYAVDFLQQAWPDDTTFNFNLIKGKKWNWYLRYLYGQGLEGLKLFIESSINRVSKASQSKAEDEDEKV 2DD5A09D9C48FC2D cenpn_chick Centromere protein N CENPN protein protein chick Gallus gallus (Chicken) Chicken MDEVIVEYIRRTVLKIPRDEIMAVLQKWGFLSEAQLQTINFRQTKEGISHSVAQLCEESSADLKQAALLDIIYNHIYPNKRVWSVYHMNKTGEETDFFDFRDFKKKFRRQIQSALINVTINFREYEDNAIWIRIAWGTPYTKPNQYKTSYVVYHSQTPYVFISASVLRSNLPLLCQAMVVASNYHDIHEMELRSHCLNSLKDIVFKRYSQNFQTNYPLQERNVLTENVDLRINDENRSEKERIYRLNQESFGNGPQPKLDFAQYKLETMFKSDPKWDVLEKKEPFRCLVKFSSPHLLESLKSLAPAGLADAPLSPLLTCIPQKARNYFKIREKKSLHPGSFVSP EE922893170EDB74 cenpw_chick Centromere protein W CENPW protein protein chick Gallus gallus (Chicken) Chicken MRRTVPRGTLRKIIKKHKPHLRLAANTDLLVHLSFLLFLHRLAEEARTNAFENKSKIIKPEHTIAAAKVILKKSRG B34E201BEBDE72F7 cenpl_chick Centromere protein L CENPL RCJMB04_9b13 protein protein chick Gallus gallus (Chicken) Chicken MAGGRPAGSAIEMEGAMRTLPSSGRPSGTGWQSRGPLFGRNYGLSPGSRLLPPRCQENADPQKMAFLLRKQWTLYSVSPLYKFSSADLKDYARMLGVFIAAEKQKGLAVDVDGELGIKVAMSGLPELRGSEQDPAAVLVQLSLRSSVSPKNSEEKLIWSGWFCCVAGDDLLENLPENFTCLPLFLVNGAESYTAIVGSWFQKTFDCCFRRLAISPLNLTWMAAMWTGCKVDKNTAATELLFSVPHLPQPLDISYAIHPEDAKALWDTVQKTPGEITQEEVDVFMECLYSHFHRHFKIHLSATKLVKVSTGIASAHCDGIIKFLQSQYLIGVLMLLTELAISQIQ AF3BA832A0DFD1BD cenpk_chick Centromere protein K CENPK protein protein chick Gallus gallus (Chicken) Chicken MSDSVCPIDAKEELLRECENIWKEMEKCQSKLTLLAAEPVPESDAKVSLLLTRMQALRAEYHQWQKRNPELISTNPEVLLLLGEEELQKVKKELEMVLSAVQLKNEKLKEDLEREQQWHDEQVQILNAFKEIEEEMKKEVVTDSEKRVFQELKNQMLELKEYKKKLMNALGEFLEEHFPLPEKNGNAKKKRYSEEPPEQVITIHEILEILLNQLMSTPHEPYVTVDDSFWPPYLELLLRSGIVLRHPEDPNRIRLEAFHE 3E297EF9D1500495 cenph-cenpt-1 2 9 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component physical association physical association Profiles did not fully overlap, suggesting that CENP-H and CENP-T may belong to separate sub-complexes Immunoblots of DT40 cell extracts fractionated on a Superose 6 gel-filtration column and probed using antibodies against anti-CENP-H or CENP-T. Fig. 1A cenpt-cug2-1 4 12 unspecified role unspecified role prey prey 7 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 7 unspecified role unspecified role prey prey association association Histones H3, H2A/H2B and H4 were also present but could not be unambiguously assigned as a single gene product Fig. 1B cenph-cenpt-2 4 9 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 11 unspecified role unspecified role prey prey 14 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 13 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey association association Profiles did not fully overlap, suggesting that CENP-H and CENP-T may belong to separate sub-complexes Fig. 1B Indeed, SDS-PAGE and mass-spectrometry analysis indicated that CENP-H IPs primarily contained CENP-H, -I, -K, -L, -M, and -N, but not CENP-T. cenpt-cug2-2 3 7 unspecified role unspecified role prey prey 12 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 physical association physical association Fig. 1C Silver staining and western blot analysis with antibodies against CENP- T confirmed clear coprecipitation of CENP-T with CUG2 cenpc-cug2-3 1 6 unspecified role unspecified role neutral component neutral component 12 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 colocalization colocalization Fig. 1D CUG2-GFP colocalized with CENP-C throughout the cell cycle (Figure 1D), demonstrating that CUG2 localizes constitutively to centromeres. cenpc-cenpa-1 1 6 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig. 5A Simultaneous detection of CENP-A and CENP-C this interaction occurred in CENP-A-deficient cells. cenpt-cenpa-1 1 8 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig. 5A Simultaneous detection of CENP-A and CENP-C or CENP-A and CENP-T cenpt-cenpa-2 3 7 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 8 unspecified role unspecified role prey prey physical association physical association Fig. 3G However, CENP-T and CENP-C coprecipitated with CENP-A when the partially digested chromatin was used. Interaction only observed when cellular chromatin is partially digested cenpc-cenpa-2 3 6 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 8 unspecified role unspecified role prey prey physical association physical association Fig. 3G However, CENP-T and CENP-C coprecipitated with CENP-A when the partially digested chromatin was used. Interaction only observed when cellular chromatin is partially digested