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/ andreasson-2005-1 The MAP kinase substrate MKS1 is a regulator of plant defense responses. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Andreasson E., Jenkins T., Brodersen P., Thorgrimsen S., Petersen NH., Zhu S., Qiu JL., Micheelsen P., Rocher A., Petersen M., Newman MA., Bjorn Nielsen H., Hirt H., Somssich I., Mattsson O., Mundy J. mundy@biobase.dk Arabidopsis cDNA library EMBO J. (0261-4189) 2005 andreasson-2005-3 The MAP kinase substrate MKS1 is a regulator of plant defense responses. in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Andreasson E., Jenkins T., Brodersen P., Thorgrimsen S., Petersen NH., Zhu S., Qiu JL., Micheelsen P., Rocher A., Petersen M., Newman MA., Bjorn Nielsen H., Hirt H., Somssich I., Mattsson O., Mundy J. mundy@biobase.dk Arabidopsis cDNA library EMBO J. (0261-4189) 2005 andreasson-2005-4 The MAP kinase substrate MKS1 is a regulator of plant defense responses. in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Andreasson E., Jenkins T., Brodersen P., Thorgrimsen S., Petersen NH., Zhu S., Qiu JL., Micheelsen P., Rocher A., Petersen M., Newman MA., Bjorn Nielsen H., Hirt H., Somssich I., Mattsson O., Mundy J. mundy@biobase.dk Arabidopsis cDNA library EMBO J. (0261-4189) 2005 andreasson-2005-2 The MAP kinase substrate MKS1 is a regulator of plant defense responses. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Andreasson E., Jenkins T., Brodersen P., Thorgrimsen S., Petersen NH., Zhu S., Qiu JL., Micheelsen P., Rocher A., Petersen M., Newman MA., Bjorn Nielsen H., Hirt H., Somssich I., Mattsson O., Mundy J. mundy@biobase.dk Arabidopsis cDNA library EMBO J. (0261-4189) 2005 andreasson-2005-5 The MAP kinase substrate MKS1 is a regulator of plant defense responses. arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Andreasson E., Jenkins T., Brodersen P., Thorgrimsen S., Petersen NH., Zhu S., Qiu JL., Micheelsen P., Rocher A., Petersen M., Newman MA., Bjorn Nielsen H., Hirt H., Somssich I., Mattsson O., Mundy J. Transgenic arabidopsis plants expressing epitope tagged MAP4 were used in this study. mundy@biobase.dk Arabidopsis cDNA library EMBO J. (0261-4189) 2005 mpk4_arath Mitogen-activated protein kinase 4 MPK4 At4g01370 F2N1.1 F2N1_2-t protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSAESCFGSSGDQSSSKGVATHGGSYVQYNVYGNLFEVSRKYVPPLRPIGRGAYGIVCAATNSETGEEVAIKKIGNAFDNIIDAKRTLREIKLLKHMDHENVIAVKDIIKPPQRENFNDVYIVYELMDTDLHQIIRSNQPLTDDHCRFFLYQLLRGLKYVHSANVLHRDLKPSNLLLNANCDLKLGDFGLARTKSETDFMTEYVVTRWYRAPELLLNCSEYTAAIDIWSVGCILGETMTREPLFPGKDYVHQLRLITELIGSPDDSSLGFLRSDNARRYVRQLPQYPRQNFAARFPNMSAGAVDLLEKMLVFDPSRRITVDEALCHPYLAPLHDINEEPVCVRPFNFDFEQPTLTEENIKELIYRETVKFNPQDSV mks1_arath Protein MKS1 MKS1 Protein MAP kinase 4 substrate 1 VQ motif-containing protein 21 VQ21 MVE11.5 At3g18690 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDPSEYFAGGNPSDQQNQKRQLQICGPRPSPLSVHKDSHKIKKPPKHPAPPPNRDQPPPYIPREPVVIYAVSPKVVHATASEFMNVVQRLTGISSGVFLESGGGGDVSPAARLASTENASPRGGKEPAARDETVEINTAMEEAAEFGGYAPGILSPSPALLPTASTGIFSPMYHQGGMFSPAIPLGLFSPAGFMSPFRSPGFTSLVASPTFADFFSHIWDQD wrk33_arath Probable WRKY transcription factor 33 WRKY33 At2g38470 T19C21.4 WRKY DNA-binding protein 33 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAASFLTMDNSRTRQNMNGSANWSQQSGRTSTSSLEDLEIPKFRSFAPSSISISPSLVSPSTCFSPSLFLDSPAFVSSSANVLASPTTGALITNVTNQKGINEGDKSNNNNFNLFDFSFHTQSSGVSAPTTTTTTTTTTTTTNSSIFQSQEQQKKNQSEQWSQTETRPNNQAVSYNGREQRKGEDGYNWRKYGQKQVKGSENPRSYYKCTFPNCPTKKKVERSLEGQITEIVYKGSHNHPKPQSTRRSSSSSSTFHSAVYNASLDHNRQASSDQPNSNNSFHQSDSFGMQQEDNTTSDSVGDDEFEQGSSIVSRDEEDCGSEPEAKRWKGDNETNGGNGGGSKTVREPRIVVQTTSDIDILDDGYRWRKYGQKVVKGNPNPRSYYKCTTIGCPVRKHVERASHDMRAVITTYEGKHNHDVPAARGSGYATNRAPQDSSSVPIRPAAIAGHSNYTTSSQAPYTLQMLHNNNTNTGPFGYAMNNNNNNSNLQTQQNFVGGGFSRAKEEPNEETSFFDSFMP wrk25_arath Probable WRKY transcription factor 25 WRKY25 At2g30250 T9D9.6 WRKY DNA-binding protein 25 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSSTSFTDLLGSSGVDCYEDDEDLRVSGSSFGGYYPERTGSGLPKFKTAQPPPLPISQSSHNFTFSDYLDSPLLLSSSHSLISPTTGTFPLQGFNGTTNNHSDFPWQLQSQPSNASSALQETYGVQDHEKKQEMIPNEIATQNNNQSFGTERQIKIPAYMVSRNSNDGYGWRKYGQKQVKKSENPRSYFKCTYPDCVSKKIVETASDGQITEIIYKGGHNHPKPEFTKRPSQSSLPSSVNGRRLFNPASVVSEPHDQSENSSISFDYSDLEQKSFKSEYGEIDEEEEQPEMKRMKREGEDEGMSIEVSKGVKEPRVVVQTISDIDVLIDGFRWRKYGQKVVKGNTNPRSYYKCTFQGCGVKKQVERSAADERAVLTTYEGRHNHDIPTALRRS mpk6_arath Mitogen-activated protein kinase 6 MPK6 At2g43790 F18O19.10 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDGGSGQPAADTEMTEAPGGFPAAAPSPQMPGIENIPATLSHGGRFIQYNIFGNIFEVTAKYKPPIMPIGKGAYGIVCSAMNSETNESVAIKKIANAFDNKIDAKRTLREIKLLRHMDHENIVAIRDIIPPPLRNAFNDVYIAYELMDTDLHQIIRSNQALSEEHCQYFLYQILRGLKYIHSANVLHRDLKPSNLLLNANCDLKICDFGLARVTSESDFMTEYVVTRWYRAPELLLNSSDYTAAIDVWSVGCIFMELMDRKPLFPGRDHVHQLRLLMELIGTPSEEELEFLNENAKRYIRQLPPYPRQSITDKFPTVHPLAIDLIEKMLTFDPRRRITVLDALAHPYLNSLHDISDEPECTIPFNFDFENHALSEEQMKELIYREALAFNPEYQQ mpk4-msk1 Yeast 2 hybrid analysis reveals interaction between MPK4 protein and the MSK1 protein 664585 N/A 664590 unspecified role unspecified role bait bait N/A 664591 unspecified role unspecified role prey prey physical association physical association false false false msk1-wrky33 Yeast 2 hybrid analysis reveals interaction between MSK1 protein and the WRKY33 protein 664585 N/A 664592 unspecified role unspecified role prey prey N/A 664591 unspecified role unspecified role bait bait physical association physical association false false false msk1-wrky25 Yeast 2 hybrid analysis reveals interaction between MSK1 protein and the WRKY25 protein 664585 N/A 664591 unspecified role unspecified role bait bait N/A 664593 unspecified role unspecified role prey prey physical association physical association false false false msk1-mpk4-1 Pulldown analysis reveals interaction between MSK1 protein and the MPK4 protein 664586 N/A 664591 unspecified role unspecified role bait bait n-terminal 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 ecoli Escherichia coli (strain K12) N/A 664590 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro physical association physical association false false false 1 msk1-wrky33-1 Pulldown analysis reveals interaction between MSK1 protein and the WRKY33 protein 664586 N/A 664591 unspecified role unspecified role bait bait n-terminal 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 ecoli Escherichia coli (strain K12) N/A 664592 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro physical association physical association false false false 1 msk1-wrky25-1 Pulldown analysis reveals interaction between MSK1 protein and the WRKY25 protein 664586 N/A 664593 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 664591 unspecified role unspecified role bait bait n-terminal 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 ecoli Escherichia coli (strain K12) physical association physical association false false false 1 mpk4-msk1-1 In vitro phosphorylation analysis of MPK4 protein and the MSK1 protein 664587 N/A 664590 enzyme enzyme neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 664591 enzyme target enzyme target substrate substrate neutral component neutral component Q8LGD5:p.Ser30Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false 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 phosphorylation phosphorylation reaction phosphorylation false false false 1 The author's state that "This indicates that MKS1 is an in vitro substrate of MPK4, and that MKS1 Ser30 and/or Ser72, both contained in the C3 truncation, may be phosphorylation sites." phosphorylation of amino acid residue serine 30 mpk4-wrky25 In vitro phosphorylation analysis of MPK4 protein and the WRKY25 protein 664587 N/A 664590 enzyme enzyme neutral component neutral component N/A 664593 enzyme target enzyme target substrate substrate neutral component neutral component phosphorylation phosphorylation reaction phosphorylation false false false 5 flagellin Phosphorylation of WRKY25 protein. mpk6-wrky25 In vitro phosphorylation analysis of MPK6 protein and the WRKY25 protein 664587 N/A 664593 enzyme target enzyme target substrate substrate neutral component neutral component N/A 664594 enzyme enzyme neutral component neutral component arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress phosphorylation phosphorylation reaction phosphorylation false false false 5 flagellin Phosphorylation of WRKY25 protein. msk1-mpk4 Yeast 2 hybrid analysis reveals interaction between MSK1 protein and the MPK4 protein 664588 N/A 664590 unspecified role unspecified role prey prey N/A 664591 unspecified role unspecified role bait bait physical association physical association false false false mpk4-msk1-2 Yeast 2 hybrid analysis reveals interaction between MPK4 protein and the MSK1 protein 664588 N/A 664591 unspecified role unspecified role prey prey N/A 664590 unspecified role unspecified role bait bait physical association physical association false false false msk1-mpk4-2 Coip analysis reveals interaction between MSK1 protein and the MPK4 protein 664589 N/A 664590 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 664591 unspecified role unspecified role bait bait physical association physical association false false false 3 The authors state that "In addition, the levels of phosphorylated MKS1 detected with a phosphoserine/phosphothreonine-specific antibody were markedly higher in wild-type plants than in mpk4 mutants" msk1-wrky25-2 Coip analysis reveals interaction between MSK1 protein and the WRKY25 protein 664589 N/A 664590 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 664593 unspecified role unspecified role prey prey physical association physical association false false false 5D The authors state that "MPK4 that phosphorylated the N-terminal regions of W25 and W33". The assay used and coordinates of this region of the WRKY25 protein are not clearly set out in the text.