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/ nishino-2012-2 CENP-T-W-S-X Forms a Unique Centromeric Chromatin Structure with a Histone-like Fold. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Nishino T., Takeuchi K., Gascoigne K.E., Suzuki A., Hori T., Oyama T., Morikawa K., Cheeseman I.M., Fukagawa T. Selenomethionine derivatives of the proteins were crystallised. tfukagaw@lab.nig.ac.jp Cell (0092-8674) 2012 Only protein-protein interactions imex curation nishino-2012-1 CENP-T-W-S-X Forms a Unique Centromeric Chromatin Structure with a Histone-like Fold. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Nishino T., Takeuchi K., Gascoigne K.E., Suzuki A., Hori T., Oyama T., Morikawa K., Cheeseman I.M., Fukagawa T. Selenomethionine derivatives of the proteins were crystallised. tfukagaw@lab.nig.ac.jp Cell (0092-8674) 2012 Only protein-protein interactions imex curation nishino-2012-3 CENP-T-W-S-X Forms a Unique Centromeric Chromatin Structure with a Histone-like Fold. chick-dt40 Gallus gallus bursa lymphoma dt40 chicken bursa lymphoma http://www.lgcpromochem-atcc.com/common/catalog/numSearch/numResults.cfm ATCC Number:CRL-2111 chicken bursa lymphoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Nishino T., Takeuchi K., Gascoigne K.E., Suzuki A., Hori T., Oyama T., Morikawa K., Cheeseman I.M., Fukagawa T. Selenomethionine derivatives of the proteins were crystallised. tfukagaw@lab.nig.ac.jp Cell (0092-8674) 2012 Only protein-protein interactions imex curation cenpw_chick Centromere protein W CENPW protein protein chick Gallus gallus (Chicken) Chicken MRRTVPRGTLRKIIKKHKPHLRLAANTDLLVHLSFLLFLHRLAEEARTNAFENKSKIIKPEHTIAAAKVILKKSRG cenpt_chick Centromere protein T CENPT protein protein chick Gallus gallus (Chicken) Chicken MDGRVGLRASRRAAPTPRVAVRSSPRHRARVREQEPVVSAMSGSGLGKENKAGSSTPLRHRPNAFSELDNATPRVMLRKIIQNQPQVSPLALQTVQLEETEDARPEPPSQRTSSTVELQLPDLVPEDASVTTFRMTRKRKKLSISEFERAADKRLPQNQAHSTLDSTLVRSLRMSVGSVMAPDTVEKRGLLRRPQNHKAIDIAAFEGGVEQNMLQIKAQDYLVDLQTSSMTGTTTIRTDAEVVLNNTELFVEPQLGEQNLLAVEPQLSDSKTSAQRSNTSYPAHEKARLEGLVSRVSTDERRTLRFSEKDLITDHEHVDGITQKTPAKQGEEEQDHSQQNDPMEQFSESEEMAGTTEHHADAEYSEHSEKKLSRKAVSQLTAAQDAGVEMEMTPSEGGVAEGTEHQDSPKAELQMAGSPGGHSPASYSLEKPGAKPLKEAVEQTGEIERGTITGVLDAAEEEATDDESDKEDHESEEISMKTPMFVHAAAYRPQPVLSPPHPVKSASPELPPQPVRAKPVPKSSGAAQRKTREPEIASSLIKQIFSHYVKTPVTRDAYKIVEKCSERYFKQISSDLEAYSQHAGRKTVEMADVELLMRRQGLVTDKMPLHVLVERHLPLEYRKLLIPIAVSGNKVIPCK cenpx_chick Centromere protein X CENPX STRA13 protein protein chick Gallus gallus (Chicken) Chicken MEEREGGFRKETVERLLRLHFRDGRTRVNGDALLLMAELLKVFVREAAARAARQAQAEDLEKVDIEHVEKVLPQLLLDFV cenps_chick Centromere protein S CENPS APITD1 protein protein chick Gallus gallus (Chicken) Chicken MEAAGGEQRELLIQRLRAAVHYTTGCLCQDVAEDKGVLFSKQTVAAISEITFRQCENFARDLEMFARHAKRSTITSEDVKLLARRSNSLLKYITQKSDELASSNMEQKEKKKKKSSAAKGRKTEENETPVTESEDSNMA Similar to Q8C4X1 cenpt_human Centromere protein T Interphase centromere complex protein 22 CENPT C16orf56 ICEN22 protein protein human Homo sapiens Human MADHNPDSDSTPRTLLRRVLDTADPRTPRRPRSARAGARRALLETASPRKLSGQTRTIARGRSHGARSVGRSAHIQASGHLEEQTPRTLLKNILLTAPESSILMPESVVKPVPAPQAVQPSRQESSCGSLELQLPELEPPTTLAPGLLAPGRRKQRLRLSVFQQGVDQGLSLSQEPQGNADASSLTRSLNLTFATPLQPQSVQRPGLARRPPARRAVDVGAFLRDLRDTSLAPPNIVLEDTQPFSQPMVGSPNVYHSLPCTPHTGAEDAEQAAGRKTQSSGPGLQKNSPGKPAQFLAGEAEEVNAFALGFLSTSSGVSGEDEVEPLHDGVEEAEKKMEEEGVSVSEMEATGAQGPSRVEEAEGHTEVTEAEGSQGTAEADGPGASSGDEDASGRAASPESASSTPESLQARRHHQFLEPAPAPGAAVLSSEPAEPLLVRHPPRPRTTGPRPRQDPHKAGLSHYVKLFSFYAKMPMERKALEMVEKCLDKYFQHLCDDLEVFAAHAGRKTVKPEDLELLMRRQGLVTDQVSLHVLVERHLPLEYRQLLIPCAYSGNSVFPAQ cenpw_human Centromere protein W CENPW C6orf173 CUG2 Cancer-up-regulated gene 2 protein protein protein human Homo sapiens Human MALSTIVSQRKQIKRKAPRGFLKRVFKRKKPQLRLEKSGDLLVHLNCLLFVHRLAEESRTNACASKCRVINKEHVLAAAKVILKKSRG cenps_human Centromere protein S CENPS APITD1 Apoptosis-inducing TAF9-like domain-containing protein 1 FAAP16 FANCM-associated histone fold protein 1 FANCM-interacting histone fold protein 1 Fanconi anemia-associated polypeptide of 16 kDa MHF1 protein protein human Homo sapiens Human MEEEAETEEQQRFSYQQRLKAAVHYTVGCLCEEVALDKEMQFSKQTIAAISELTFRQCENFAKDLEMFARHAKRTTINTEDVKLLARRSNSLLKYITDKSEEIAQINLERKAQKKKKSEDGSKNSRQPAEAGVVESEN cenpx_human Centromere protein X CENPX FAAP10 FANCM-associated histone fold protein 2 FANCM-interacting histone fold protein 2 Fanconi anemia-associated polypeptide of 10 kDa MHF2 Retinoic acid-inducible gene D9 protein homolog STRA13 Stimulated by retinoic acid gene 13 protein homolog protein protein human Homo sapiens Human MEGAGAGSGFRKELVSRLLHLHFKDDKTKVSGDALQLMVELLKVFVVEAAVRGVRQAQAEDALRVDVDQLEKVLPQLLLDF cenpw-cenpt-1 1268646 n/a 1268649 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1268650 unspecified role unspecified role neutral component neutral component purified purified F1NPG5:p.[Met590Leu;Met607Leu] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 direct interaction direct interaction false false false Fig. 4B, 4E, Supp. Fig. 4A apitd1-cenpt-2 1268646 n/a 1268651 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1268652 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1268649 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1268650 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false F1NPG5:p.[Met590Leu;Met607Leu] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 physical association physical association false false false Fig. 4B,Supp. Fig. 4A CenpS lacking the C-terminal 33 amino acids also bound in the complex. cenps-cenpx 1268646 n/a 1268651 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 n/a 1268652 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Fig. 4B, 4E, Supp. Fig. 2B, 3A, 4A Mutant CENP-S-X complexes (containing CENP-S [R15A, K41A, K70A] or CENP-X [R9A, R27A, K62A]; bind but are defective in DNA binding. cenps-cenpx 1268646 n/a 1268651 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1268652 unspecified role unspecified role neutral component neutral component purified purified E1BSW7:p.[Phe65Glu;His68Glu;Leu81Arg;Arg84Leu] mutation mutation 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 certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 direct interaction direct interaction false false false Supp Fig 2B, 2C Mutations cause protein to dimerize rather than tetramize. Loss of the C-terminal 33 amino acids also prevented tetramerization. Tetramerization of the CENP-S-X complex is also required for full DNA binding activity cenpt-cenpw 1268646 n/a 1268653 unspecified role unspecified role neutral component neutral component purified purified region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 1268654 unspecified role unspecified role neutral component neutral component purified purified region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Supp. Fig. 4B apitd1-cenpw-1 1268646 n/a 1268655 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1268656 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 direct interaction direct interaction false false false Supp. Fig. 4B apitd1-cenpt-1 1268646 n/a 1268655 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1268654 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1268653 unspecified role unspecified role neutral component neutral component purified purified region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1268656 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 physical association physical association false false false Supp. Fig. 4B The N-terminus of CENPT is not required for bindingto the complex. cenpt-cenpw-1 1268647 n/a 1268649 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1268650 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false F1NPG5:p.[Met590Leu;Met607Leu] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 direct interaction direct interaction false false false Rworking =22.0%, Rfree = 26.9% 2.2A Fig. 1A, Supp. Table 1 Selenomethionine derivatives of the proteins were utilised. cenps-cenpx-1 1268647 n/a 1268651 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 n/a 1268652 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false 2.15A Rworking 18.6%, Rfree 21.8% Fig. 2A, Supp. Table 2 Selenomethionine derivatives of the proteins were utilized. gga_2263-gga_2856-1 1268648 n/a 1268650 unspecified role unspecified role bait bait n/a 1268652 unspecified role unspecified role prey prey E1BSW7:p.[Phe65Glu;His68Glu;Leu81Arg;Arg84Glu] mutation mutation 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 certain certain sequence position certain certain certain sequence position certain false region rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Supp. 3B CENPS mutant defective in DNA binding still bound CENPT. cenpt-apitd1 1268648 n/a 1268650 unspecified role unspecified role bait bait n/a 1268652 unspecified role unspecified role prey prey physical association physical association false false false Supp. 3B CENPS mutant defective in DNA binding still bound CENPT.