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/ miranda-2005-1 The yeast DASH complex forms closed rings on microtubules. ecoli Escherichia coli (strain K12) chromatography chromatography technology column chromatography chromatography weight by comassie molecular weight estimation by coomasie staining weight by comassie Miranda JJ., De Wulf P., Sorger PK., Harrison SC. All ten subunits were coexpressed from one vector and purifed using a HiPrep 16/60 Sephacryl S-500 HR column. Identity of some participants confirmed by western blot harrison@crystal.harvard.edu Nat. Struct. Mol. Biol. (1545-9993) 2005 Only protein-protein interactions imex curation miranda-2005-2 The yeast DASH complex forms closed rings on microtubules. ecoli Escherichia coli (strain K12) molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight identificat confirmation by molecular weight weight identificat Miranda JJ., De Wulf P., Sorger PK., Harrison SC. All ten subunits were coexpressed from one vector and purifed using a HiPrep 16/60 Sephacryl S-500 HR column. Identity of some participants confirmed by western blot harrison@crystal.harvard.edu Nat. Struct. Mol. Biol. (1545-9993) 2005 Only protein-protein interactions imex curation miranda-2005-3 The yeast DASH complex forms closed rings on microtubules. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight identificat confirmation by molecular weight weight identificat Miranda JJ., De Wulf P., Sorger PK., Harrison SC. All ten subunits were coexpressed from one vector and purifed using a HiPrep 16/60 Sephacryl S-500 HR column. Identity of some participants confirmed by western blot harrison@crystal.harvard.edu Nat. Struct. Mol. Biol. (1545-9993) 2005 Only protein-protein interactions imex curation miranda-2005-4 The yeast DASH complex forms closed rings on microtubules. ecoli Escherichia coli (strain K12) cosedimentation cosedimentation weight identificat confirmation by molecular weight weight identificat Miranda JJ., De Wulf P., Sorger PK., Harrison SC. All ten subunits were coexpressed from one vector and purifed using a HiPrep 16/60 Sephacryl S-500 HR column. Identity of some participants confirmed by western blot harrison@crystal.harvard.edu Nat. Struct. Mol. Biol. (1545-9993) 2005 Only protein-protein interactions imex curation miranda-2005-5 The yeast DASH complex forms closed rings on microtubules. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast cosedimentation cosedimentation weight identificat confirmation by molecular weight weight identificat Miranda JJ., De Wulf P., Sorger PK., Harrison SC. All ten subunits were coexpressed from one vector and purifed using a HiPrep 16/60 Sephacryl S-500 HR column. Identity of some participants confirmed by western blot harrison@crystal.harvard.edu Nat. Struct. Mol. Biol. (1545-9993) 2005 Only protein-protein interactions imex curation dad4_yeast DASH complex subunit DAD4 Outer kinetochore protein DAD4 DUO1 and DAM1-interacting protein 4 Helper of ASK1 protein 2 DAD4 HSK2 YDR320C-A protein protein yeast Saccharomyces cerevisiae Baker's yeast MENPHEQVQANILSRIIGNVKRLNESVAILNQELVTINNRNKNLEIMGAICDNYHSSVQFNLEATNNKKPPL dad3_yeast DASH complex subunit DAD3 Outer kinetochore protein DAD3 DUO1 and DAM1-interacting protein 3 DAD3 YBR233W-A protein protein yeast Saccharomyces cerevisiae Baker's yeast MEHNLSPLQQEVLDKYKQLSLDLKALDETIKELNYSQHRQQHSQQETVSPDEILQEMRDIEVKIGLVGTLLKGSVYSLILQRKQEQESLGSNSK duo1_yeast DASH complex subunit DUO1 YGL061C DUO1 Outer kinetochore protein DUO1 Death upon overproduction protein 1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSEQSQLDDSTIDKLIPQIFNEMRSNLNNTTNKFPKSTGGGASDNISANSNSIRSFNSITTQSLLKESESLDKITAMIKNVTAALKNNLPVYVNQVHEVCKSTNSILDSWINIHSQAGYIHKLMSDQTYLKLINDRLHNENVNTNDEDGSTLHNVIALKKKEILDLRQKLENRKGEKDAAPAKPPNQGLNPRYGVQSGRRPVPSAGISNNGRVRKTHVPASKRPSGIPRVTNRWTKPTASSSRKMFR 787F8AF869E3C978 spc34_yeast DASH complex subunit SPC34 YKR037C SPC34 34 kDa spindle pole component protein Outer kinetochore protein SPC34 protein protein yeast Saccharomyces cerevisiae Baker's yeast MGESLDRCIDDINRAVDSMSTLYFKPPGIFHNAILQGASNKASIRKDITRLIKDCNHDEAYLLFKVNPEKQSVSRRDGKEGVFDYVIKRDTDMKRNRRLGRPGEKPIIHVPKEVYLNKDRLDLNNKRRRTATTSGGGLNGFIFDTDLIGSSVISNSSSGTFKALSAVFKDDPQIQRLLYALENGSVLMEEESNNQRRKTIFVEDFPTDLILKVMAEVTDLWPLTEFKQDYDQLYHNYEQLSSKLRFIKKEVLLQDDRLKTMSQYHPSSSHDVAKIIRKEKDEIRRLEMEIANLQE C1061B49F6D4D308 dad1_yeast DASH complex subunit DAD1 DAD1 Outer kinetochore protein DAD1 DUO1 and DAM1-interacting protein 1 YDR016C protein protein yeast Saccharomyces cerevisiae Baker's yeast MMASTSNDEEKLISTTDKYFIEQRNIVLQEINETMNSILNGLNGLNISLESSIAVGREFQSVSDLWKTLYDGLESLSDEAPIDEQPTLSQSKTK dad2_yeast DASH complex subunit DAD2 YKR083C YKR403 HSK1 DAD2 Outer kinetochore protein DAD2 DUO1 and DAM1-interacting protein 2 Helper of ASK1 protein 1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MDSIDEQIAIKRKELQSLQKITSLTDGLKIQLTELNEQIKEMGMNADSVAQLMNNWDSIINNISQASLGLLQYAEGDYEIGPWKDSKKKESEQSNETGLEAQENDKNDEDNDEDEDLVPLPETMVRIRVDGNE 82730D4AA5B66951 spc19_yeast DASH complex subunit SPC19 SPC19 19 kDa spindle pole component protein Outer kinetochore protein SPC19 YDR201W protein protein yeast Saccharomyces cerevisiae Baker's yeast MTDALEQSVLALEGTVSVLKDSVESLKCANEPSTNLASTMLQTKRVFRLVPEYDVERSKLDLIEEVEPLVRTLGDKLRKSMGRMQRELDTLQQTYELNDLRLKKNISMDDDDALNSPDMGQEYEGRDADDVVMMASSTNEELEELKKLKEKKKQLENKLEILKQK 74615FE2186DAFE6 ask1_yeast DASH complex subunit ASK1 YKL052C YKL306 ASK1 Outer kinetochore protein ASK1 Associated with spindles and kinetochores protein 1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MDSASKEETLEKLDQEITVNLQKIDSNLSFCFHKITQDIIPHVATYSEICERIMDSTEWLGTMFQETGLVNLQANAAAPVGNAPVKSLVSNNVGIFPTSAEEASRQSQTDNGPNEADSAVHVNRDVHSMFNNDSIDDFHTANITSTGQILKLPDSSDEDTGSEAVPSREQTDLTGEGHGGADDEQDESTIQRQSRKRKISLLLQQQYGSSSSMVPSPIVPNKMRKQLAHEEHINNDGDNDDENSNNIESSPLKQGHHHPKGQADDNNEGPDEEESTKEVPKPGTIIHFSTNR dam1_yeast DASH complex subunit DAM1 YGR113W G6153 DAM1 Outer kinetochore protein DAM1 DUO1 and MPS1-interacting protein 1 Kinetochore assembly protein DAM1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSEDKAKLGTTRSATEYRLSIGSAPTSRRSSMGESSSLMKFADQEGLTSSVGEYNENTIQQLLLPKIRELSDSIITLDSNFTRLNFIHESLADLNESLGSLLYGIMSNSWCVEFSQAPHDIQDDLIAIKQLKSLEDEKNNLVMELSNMERGIKRKKDEQGENDLAKASQNKQFNQPLFPSSQVRKYRSYDNRDKRKPSKIGNNLQVENEEDYEDDTSSEASFVLNPTNIGMSKSSQGHVTKTTRLNNNTNSKLRRKSILHTIRNSIASGADLPIENDNVVNLGDLHPNNRISLGSGAARVVNGPVTKNRNSMFSGRAERKPTESRHSVAKKTEKKINTRPPFR F02DB6EE09BC2284 hsk3_yeast DASH complex subunit HSK3 Outer kinetochore protein HSK3 Helper of ASK1 protein 3 HSK3 YKL138C-A protein protein yeast Saccharomyces cerevisiae Baker's yeast MNANKQRQYNQLAHELRELQTNLQETTKQLDIMSKQCNENLVGQLGKVHGSWLIGSYIYYMEQMLGKTQ ask1-dad1-1 Components of the DASH complex identified by chromatography 659371 N/A 659376 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659377 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659378 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659379 unspecified role unspecified role neutral component neutral component c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false Stoichiometry: 1.0 N/A 659380 unspecified role unspecified role neutral component neutral component c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false Stoichiometry: 1.0 N/A 659381 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659382 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659383 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659384 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659385 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 physical association physical association false false false Supp. Figure 1 ask1-dad1-2 Components of the DASH complex identified by chromatography 659372 N/A 659377 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659376 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659383 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659381 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659385 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659378 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659379 unspecified role unspecified role neutral component neutral component c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false Stoichiometry: 1.0 N/A 659384 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659382 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659380 unspecified role unspecified role neutral component neutral component c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false Stoichiometry: 1.0 physical association physical association false false false Figure 1a M.W.= 211 +/- 16 kDa, ask1-dad1-3 Components of the DASH complex identified by chromatography 659373 N/A 659376 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659377 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659382 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659383 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659379 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659384 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659378 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659380 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659381 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659385 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 physical association physical association false false false Figure 1a M.W.= 211 +/- 16 kDa, ask1-dad1-4 Components of the DASH complex identified by sedimentation velocity ultracentrifugation 659374 N/A 659376 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659377 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659378 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659379 unspecified role unspecified role neutral component neutral component c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false Stoichiometry: 1.0 N/A 659380 unspecified role unspecified role neutral component neutral component c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false Stoichiometry: 1.0 N/A 659381 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659384 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659385 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659382 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659383 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 physical association physical association false false false Figure 1b M.W.= 211 +/- 16 kDa, ask1-dad1-5 Components of the DASH complex identified by sedimentation velocity ultracentrifugation 659375 N/A 659377 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659376 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659379 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659378 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659381 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659384 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659382 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659383 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659385 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 659380 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 physical association physical association false false false Figure 1b M.W.= 211 +/- 16 kDa,