UniProt UniProt http://www.uniprot.org jaspersen-2006-2 The Sad1-UNC-84 homology domain in Mps3 interacts with Mps2 to connect the spindle pole body with the nuclear envelope. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Jaspersen SL., Martin AE., Glazko G., Giddings TH., Morgan G., Mushegian A., Winey M. Competition experiment where the binding of Alexa-labelled, wild type MPS3 protein (457-617) was assessed in the presence of unlabelled wild type MPS3 protein or MPS3 protein mutants. slj@stowers-institute.org J. Cell Biol. (0021-9525) 2006 Only protein-protein interactions imex curation jaspersen-2006-1 The Sad1-UNC-84 homology domain in Mps3 interacts with Mps2 to connect the spindle pole body with the nuclear envelope. in vitro In vitro biochemical biochemical tag fluorescence tag visualisation by fluorescence tag fluorescence Jaspersen SL., Martin AE., Glazko G., Giddings TH., Morgan G., Mushegian A., Winey M. Competition experiment where the binding of Alexa-labelled, wild type MPS3 protein (457-617) was assessed in the presence of unlabelled wild type MPS3 protein or MPS3 protein mutants. slj@stowers-institute.org J. Cell Biol. (0021-9525) 2006 Only protein-protein interactions imex curation jaspersen-2006-3 The Sad1-UNC-84 homology domain in Mps3 interacts with Mps2 to connect the spindle pole body with the nuclear envelope. in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Jaspersen SL., Martin AE., Glazko G., Giddings TH., Morgan G., Mushegian A., Winey M. Competition experiment where the binding of Alexa-labelled, wild type MPS3 protein (457-617) was assessed in the presence of unlabelled wild type MPS3 protein or MPS3 protein mutants. slj@stowers-institute.org J. Cell Biol. (0021-9525) 2006 Only protein-protein interactions imex curation jaspersen-2006-4 The Sad1-UNC-84 homology domain in Mps3 interacts with Mps2 to connect the spindle pole body with the nuclear envelope. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Jaspersen SL., Martin AE., Glazko G., Giddings TH., Morgan G., Mushegian A., Winey M. Competition experiment where the binding of Alexa-labelled, wild type MPS3 protein (457-617) was assessed in the presence of unlabelled wild type MPS3 protein or MPS3 protein mutants. slj@stowers-institute.org J. Cell Biol. (0021-9525) 2006 Only protein-protein interactions imex curation cdc31_yeast Cell division control protein 31 YOR257W CDC31 DSK1 Nucleoporin CDC31 Nuclear pore protein CDC31 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSKNRSSLQSGPLNSELLEEQKQEIYEAFSLFDMNNDGFLDYHELKVAMKALGFELPKREILDLIDEYDSEGRHLMKYDDFYIVMGEKILKRDPLDEIKRAFQLFDDDHTGKISIKNLRRVAKELGETLTDEELRAMIEEFDLDGDGEINENEFIAICTDS 25A546E76E1EF520 mps3_yeast Spindle pole body assembly component MPS3 J1310 J1315 YJL018W YJL019W MPS3 NEP98 Monopolar spindle protein 3 98 kDa nuclear envelope protein protein protein yeast Saccharomyces cerevisiae Baker's yeast MNNSNEHRREEAGAANEQMPYNKAVKSAYADVLKDKMNREQEISLRAIKKGIYTDGGETDNYDMDKENDSAYEMFKKNLDFPLDQHNDDDDDDPYIEDNGQETDGYSDEDYTDEADKSFIEDSDSDSYDLESNSDFEENLESSGEAKKLKWRTYIFYGGLFFVFYFFGSFLMTTVKNNDLESHSSGATSSPGKSFSNLQKQVNHLYSELSKRDEKHSSELDKTVKIIVSQFEKNIKRLLPSNLVNFENDINSLTKQVETISTSMSELQRRNHKFTVENVTQWQDQLVKQLDTHLPQEIPVVINNSSSLLIIPELHNYLSALISDVIESPGIGTAGSAESRWEYDLNRYVKEILSNELQYIDKDYFIQEMNRRLQSNKQEIWEEITNRLETQQQQQQQQVQQDYSNVPQQYSSILMKRLIHQIYNSNQHQWEDDLDFATYVQGTKLLNHLTSPTWRQGSGVQPIELLTDSKQSSSTYWQCENEPGCSWAIRFKTPLYLTKISYMHGRFTNNLHIMNSAPRLISLYVKLSQTKEIKALQTLANQYGFGQHHKRDRNYIKIAKFEYRLTDSRIRQQMYLPPWFIQLKPLVRSIVFQVDENYGNKKFISLRKFIINGVTPQDLQIIENNEFPVLLGDTPEYGVTQNTDEGKRKVLLSKPPYASSSTSTKFHPASNVPSFGQDELDQ mps2_yeast Monopolar spindle protein 2 YGL075C MPS2 MMC1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSNGAFDAIFEYAWGQIDKPISGDFIYGKDLPKLIEIIENIFQKAQKSGSYELRLPLFSEINKDLFRTFSNTKTFFKIHKEEFDDIFFNLVNHPLREILENAFIGVDSIPSDFIVSMNLNSPSKFLVENKNKNTEGAGISTPRKKLTESPIKLLSRNNIGKALEVQVEELKRELTAKQSLLQENERQVSELKIRLETYQEKYASIQQRFSDLQKARQVEDNQNSSRTSDPGSPLVTGIDQKAILEEFRRRLQRQTDTISFLKDQIRRERGLNCSNDKVSHSKRKHATTDGDGTFKNFISAVPSNIWVKATIRIIVCFALLAGVLPYIRKYVYAHDTPSQNSRLQLSWWENSGILSKIVWFFEDQTDLETEYRSNANVDDAYSRVFGI mps3-cdc31-2 794164 n/a 794168 unspecified role unspecified role prey prey n/a 794169 unspecified role unspecified role bait bait c-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Fig 4C mps2-mps3-2 794164 n/a 794169 unspecified role unspecified role prey prey n/a 794170 unspecified role unspecified role bait bait n-terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Fig 4D mps2-mps3-1 794165 n/a 794170 unspecified role unspecified role bait bait n-terminus mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) n/a 794169 unspecified role unspecified role prey prey P47069:p.Trp477Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region alexa label alexa label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P47069:p.Trp487Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P47069:p.[Gln572Leu;Gln573Leu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) physical association physical association false false false Fig 4A, 4B mps2-mps3-3 794166 n/a 794169 unspecified role unspecified role prey prey n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) n/a 794170 unspecified role unspecified role bait bait region binding region binding-associated region binding region mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false n-terminus mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) physical association physical association false false false Fig 5D mps3-mps2-1 794167 n/a 794170 unspecified role unspecified role prey prey c-terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 794169 unspecified role unspecified role bait bait c-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Fig 4C