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/ Regulation of MBK-2/Dyrk kinase by dynamic cortical anchoring during the oocyte-to-zygote transition. Regulation of MBK-2/Dyrk kinase by dynamic cortical anchoring during the oocyte-to-zygote transition. Curr. Biol. (0960-9822) 2007 imex curation Stitzel ML., Cheng KC., Seydoux G. gseydoux@jhmi.edu 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Stitzel ML., Cheng KC., Seydoux G. gseydoux@jhmi.edu Accepted 2010-FEB-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Curr. Biol. (0960-9822) 2007 jyoti Only protein-protein interactions imex curation Regulation of MBK-2/Dyrk kinase by dynamic cortical anchoring during the oocyte-to-zygote transition. Regulation of MBK-2/Dyrk kinase by dynamic cortical anchoring during the oocyte-to-zygote transition. Curr. Biol. (0960-9822) 2007 imex curation Stitzel ML., Cheng KC., Seydoux G. gseydoux@jhmi.edu 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions caeel-oocyte Caenorhabditis elegans oocyte oocyte Oocyte fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Stitzel ML., Cheng KC., Seydoux G. gseydoux@jhmi.edu Accepted 2010-FEB-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Curr. Biol. (0960-9822) 2007 jyoti Only protein-protein interactions imex curation Regulation of MBK-2/Dyrk kinase by dynamic cortical anchoring during the oocyte-to-zygote transition. Regulation of MBK-2/Dyrk kinase by dynamic cortical anchoring during the oocyte-to-zygote transition. Curr. Biol. (0960-9822) 2007 imex curation Stitzel ML., Cheng KC., Seydoux G. gseydoux@jhmi.edu 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions caeel Caenorhabditis elegans anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Stitzel ML., Cheng KC., Seydoux G. gseydoux@jhmi.edu Accepted 2010-FEB-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Curr. Biol. (0960-9822) 2007 jyoti Only protein-protein interactions imex curation Regulation of MBK-2/Dyrk kinase by dynamic cortical anchoring during the oocyte-to-zygote transition. Regulation of MBK-2/Dyrk kinase by dynamic cortical anchoring during the oocyte-to-zygote transition. Curr. Biol. (0960-9822) 2007 imex curation Stitzel ML., Cheng KC., Seydoux G. gseydoux@jhmi.edu 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions caeel-embryo Caenorhabditis elegans embryo embryo Embryo Defined as the development stage prior to the initiation of tissue differentiation. fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Stitzel ML., Cheng KC., Seydoux G. Tagged proteins were immunostained with an anti-tag antibody. gseydoux@jhmi.edu Accepted 2010-FEB-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Curr. Biol. (0960-9822) 2007 jyoti Only protein-protein interactions imex curation Regulation of MBK-2/Dyrk kinase by dynamic cortical anchoring during the oocyte-to-zygote transition. Regulation of MBK-2/Dyrk kinase by dynamic cortical anchoring during the oocyte-to-zygote transition. Curr. Biol. (0960-9822) 2007 imex curation Stitzel ML., Cheng KC., Seydoux G. gseydoux@jhmi.edu 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions caeel Caenorhabditis elegans anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Stitzel ML., Cheng KC., Seydoux G. gseydoux@jhmi.edu Accepted 2010-FEB-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Curr. Biol. (0960-9822) 2007 jyoti Only protein-protein interactions imex curation Regulation of MBK-2/Dyrk kinase by dynamic cortical anchoring during the oocyte-to-zygote transition. Regulation of MBK-2/Dyrk kinase by dynamic cortical anchoring during the oocyte-to-zygote transition. Curr. Biol. (0960-9822) 2007 imex curation Stitzel ML., Cheng KC., Seydoux G. gseydoux@jhmi.edu 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Stitzel ML., Cheng KC., Seydoux G. gseydoux@jhmi.edu Accepted 2010-FEB-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Curr. Biol. (0960-9822) 2007 jyoti Only protein-protein interactions imex curation mbk2_caeel Dual specificity tyrosine-phosphorylation-regulated kinase mbk-2 mbk-2 F49E11.1 Minibrain Kinase 2 Dual specificity Yak1-related kinase mbk-2 protein protein caeel Caenorhabditis elegans MAALASFTRNSRSYGQQPIDVTQQGQRDRSVMSLDAQGRSVSHECPTSTTLVRQLYLPQIPQSASFAAAPTSFSGASSSSSNHHHPVYHSQNSLPPNLLGSSQNSASSNSLVQGHRNPALGSGNTLTRSYHQPSSTNSSTNNLYGPLGTISRDLKQSIRDISPPVINSSANPHLVNYVQTSSFDNGSYEFPSGQAQQQRRLGGSQQHLAPLQQTASSLYSNPQSSSSQLLGQQQAVRPNYAYQQSLPRQQHINSHQTQAFFGTVRGPTNSTNIVTPLRASKTMIDVLAPVRDTVAAQATGLPNVGTSSSNGSSNSSSGVGSGGSGSLMTQSIGGPNKHLSASHSTLNTASTHDMMHSKIPKSPSNESLSRSHTSSSGGSQGGHNSNSGSNSGFRPEDAVQTFGAKLVPFEKNEIYNYTRVFFVGSHAKKQAGVIGGANNGGYDDENGSYQLVVHDHIAYRYEVLKVIGKGSFGQVIKAFDHKYQQYVALKLVRNEKRFHRQADEEIRILDHLRRQDSDGTHNIIHMLDYFNFRNHKCITFELLSINLYELIKRNKFQGFSLMLVRKFAYSMLLCLDLLQKNRLIHCDLKPENVLLKQQGRSGIKVIDFGSSCFDDQRIYTYIQSRFYRAPEVILGTKYGMPIDMWSLGCILAELLTGYPLLPGEDENDQLALIIELLGMPPPKSLETAKRARTFITSKGYPRYCTATSMPDGSVVLAGARSKRGKMRGPPASRSWSTALKNMGDELFVDFLKRCLDWDPETRMTPAQALKHKWLRRRLPNPPRDGLESMGGLADHEKKTETLPNIDSNANILMRKKF 74903C85CB68C428 egg3_caeel Protein tyrosine phosphatase-like protein egg-3 egg-3 Egg sterile protein 3 F44F4.2 protein protein caeel Caenorhabditis elegans MRTSDSHLPLSNLARSDSIEFKDAVINEKWVHSANRRKGHLTPKAPEKKSGWFGGQKSEEELLMERVEQHELEELQTFIAQKLFRVDGIICDEETRILLVEKLMNAKEYPTINHDELAHRYGSSMAGWLRDRLVPSMSDCSSVLQRAAAEFYQNKMSDPLCNWGQLNPEHVSMVAARIAKFSEEMSSKVKWSLLVEPGKFSCHLTEFVQEFNRLDRMFVSNELSDEESLQTAFNANYLTKARSKMVPCAEFSRVKLNDGLGRLDDRNELRNGMFSDEHEFLQEEGYTAKSTYGTTDFIHANYVKGGPLLNTFICAQAPLKNTQEDFWRMVFQEKCQFIVMLNSAVDSSTLGPLDSANRNHCPYYWPRAENESLRFGSFHITCMKVDSKADPLFTITKLKVQKVGGNLLDAEFDEELFLEHWQWDWQYLGDVHWPFRVLRKARQLSTPTIVQCIDGCSKSGTLVSIETALMHFIRGSPITKSLILQSCVFVRLQRRLSVSSVLLYLFIYRVILRWIEPYVNKWYHRAALGLRFKSIGFIQKYNAMIQEFSRITPAY 587DA01F0CC92E94 dyrktide_peptide Dyrktide peptide protein protein chemical synthesis Chemical synthesis (Chemical synthesis) RRRFRPASPLRGPPK An amino acid sequence which is not represented in UniProt 9C6720A4FDDAEAEF mbk2-egg3-1 2 7 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization As predicted from the GFP fusions, we found that endogenous MBK-2 and EGG-3 colocalize on the cortex of growing oocytes (Figure 2A). Fig. 2A mbk2-egg3-2 4 7 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 8 unspecified role unspecified role neutral component neutral component colocalization colocalization We found, however, that two MBK-2 fusions (GFP:MBK-2 and MBK-2:HIS) colocalize with endogenous EGG-3 on the cortex in metaphase I embryos (Figure 2B; Figure S1C) and in discrete cytoplasmic puncta during the second meiotic division (Figure 2C; Figure S1C) Fig. 2B, 2C MBK-2 colocalizes with endogenous EGG-3 on the cortex in metaphase I embryos and in discrete cytoplasmic puncta during the second meiotic division. mbk2-egg3-3 4 7 unspecified role unspecified role neutral component neutral component 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 8 unspecified role unspecified role neutral component neutral component colocalization colocalization We found, however, that two MBK-2 fusions (GFP:MBK-2 and MBK-2:HIS) colocalize with endogenous EGG-3 on the cortex in metaphase I embryos (Figure 2B; Figure S1C) and in discrete cytoplasmic puncta during the second meiotic division (Figure 2C; Figure S1C) Supp. Fig. 1C MBK-2 colocalizes with endogenous EGG-3 on the cortex in metaphase I embryos and in discrete cytoplasmic puncta during the second meiotic division. mbk2-egg3-4 5 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association We detected EGG-3 in MBK-2 immunoprecipitates and MBK-2 in EGG-3 immunoprecipitates Fig. 3A mbk2-egg3-5 5 7 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association We detected EGG-3 in MBK-2 immunoprecipitates and MBK-2 in EGG-3 immunoprecipitates Fig. 3B mbk2-egg3-6 3 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait 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 physical association physical association GFP antibodies could immunoprecipitate MBK-2 in extracts from worms expressing GFP:EGG-3 but not in extracts from worms expressing GFP alone Fig. 3C mbk2-egg3-7 6 8 unspecified role unspecified role bait bait n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association We found that GST:EGG-3-coupled beads could pull down FLAG:MBK-2 from E. coli extracts and, conversely, that GST:MBK-2 could pull down FLAG:EGG-3 Fig. 3D mbk2-egg3-8 6 8 unspecified role unspecified role prey prey n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role bait bait n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association We found that GST:EGG-3-coupled beads could pull down FLAG:MBK-2 from E. coli extracts and, conversely, that GST:MBK-2 could pull down FLAG:EGG-3 Fig. 3D mbk2-dyrktide-1 1 7 enzyme enzyme 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 Q9XTF3:p.Lys490Arg Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K R Wed May 02 16:03:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys490arg' Tue Jun 30 18:35:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys490Arg' caeel Caenorhabditis elegans 9 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) phosphorylation phosphorylation reaction phosphorylation Kinase assay comparing the kinase activity of GFP:MBK-2 immunoprecipitated from wild-type or mat-1(ax227) worms against a synthetic peptide (DYRKtide). Fig. 5H Phosphorylation of Dyrktide