DIP Database of Interacting Proteins 611 Young Drive East; Los Angeles CA 90095; USA http://dip.doe-mbi.ucla.edu/ dip@mbi.ucla.edu 611 Young Drive East; Los Angeles CA 90095; USA http://dip.doe-mbi.ucla.edu/ bethke-2009-1 Flg22 regulates the release of an ethylene response factor substrate from MAP kinase 6 in Arabidopsis thaliana via ethylene signaling. arath-mesophyll Arabidopsis thaliana mesophyll mesophyll Mesophyll of plant leaf The photosynthetic parenchyma between the epidermal layers of a foliage leaf. fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Bethke G., Unthan T., Uhrig JF., Poschl Y., Gust AA., Scheel D., Lee J. The authors or references in the text do not supply enough information to unambiguously identify the myelin basic protein used in this study. Arabidopsis mesophyll protoplasts were prepared as described in Asai et al., 2002. jlee@ipb-halle.de, dscheel@ipb-halle.de Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2009 IMEx Only protein-protein interactions imex curation bethke-2009-2 Flg22 regulates the release of an ethylene response factor substrate from MAP kinase 6 in Arabidopsis thaliana via ethylene signaling. in vitro In vitro protein kinase assay protein kinase assay autoradiography autoradiography Bethke G., Unthan T., Uhrig JF., Poschl Y., Gust AA., Scheel D., Lee J. The authors or references in the text do not supply enough information to unambiguously identify the myelin basic protein used in this study. Arabidopsis mesophyll protoplasts were prepared as described in Asai et al., 2002. jlee@ipb-halle.de, dscheel@ipb-halle.de Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2009 IMEx Only protein-protein interactions imex curation bethke-2009-3 Flg22 regulates the release of an ethylene response factor substrate from MAP kinase 6 in Arabidopsis thaliana via ethylene signaling. arath-mesophyll Arabidopsis thaliana mesophyll mesophyll Mesophyll of plant leaf The photosynthetic parenchyma between the epidermal layers of a foliage leaf. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Bethke G., Unthan T., Uhrig JF., Poschl Y., Gust AA., Scheel D., Lee J. The authors or references in the text do not supply enough information to unambiguously identify the myelin basic protein used in this study. Arabidopsis mesophyll protoplasts were prepared as described in Asai et al., 2002. jlee@ipb-halle.de, dscheel@ipb-halle.de Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2009 IMEx Only protein-protein interactions imex curation bethke-2009-4 Flg22 regulates the release of an ethylene response factor substrate from MAP kinase 6 in Arabidopsis thaliana via ethylene signaling. arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Bethke G., Unthan T., Uhrig JF., Poschl Y., Gust AA., Scheel D., Lee J. The authors or references in the text do not supply enough information to unambiguously identify the myelin basic protein used in this study. Arabidopsis mesophyll protoplasts were prepared as described in Asai et al., 2002. jlee@ipb-halle.de, dscheel@ipb-halle.de Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2009 IMEx Only protein-protein interactions imex curation ef104_arath Ethylene-responsive transcription factor ERF104 ERF104 K11J9.13 At5g61600 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MATKQEALAIDFISQHLLTDFVSMETDHPSLFTNQLHNFHSETGPRTITNQSPKPNSTLNQRKPPLPNLSVSRTVSTKTEKEEEERHYRGVRRRPWGKYAAEIRDPNKKGCRIWLGTYDTAVEAGRAYDQAAFQLRGRKAILNFPLDVRVTSETCSGEGVIGLGKRKRDKGSPPEEEKAARVKVEEEESNTSETTEAEVEPVVPLTPSSWMGFWDVGAGDGIFSIPPLSPTSPNFSVISVT mpk6_arath Mitogen-activated protein kinase 6 MPK6 At2g43790 F18O19.10 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDGGSGQPAADTEMTEAPGGFPAAAPSPQMPGIENIPATLSHGGRFIQYNIFGNIFEVTAKYKPPIMPIGKGAYGIVCSAMNSETNESVAIKKIANAFDNKIDAKRTLREIKLLRHMDHENIVAIRDIIPPPLRNAFNDVYIAYELMDTDLHQIIRSNQALSEEHCQYFLYQILRGLKYIHSANVLHRDLKPSNLLLNANCDLKICDFGLARVTSESDFMTEYVVTRWYRAPELLLNSSDYTAAIDVWSVGCIFMELMDRKPLFPGRDHVHQLRLLMELIGTPSEEELEFLNENAKRYIRQLPPYPRQSITDKFPTVHPLAIDLIEKMLTFDPRRRITVLDALAHPYLNSLHDISDEPECTIPFNFDFENHALSEEQMKELIYREALAFNPEYQQ pdf1_2_arath_gene PDF1.2/ Defensin-like protein 16 Arabidopsis Gene gene gene arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress mpk6-erf104-3 830243 n/a 830247 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 830248 unspecified role unspecified role neutral component neutral component region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false Figure 1 flg22 peptide (QRLSTGSRINSAKDDAAGLQIA) Kinase inactive MPK6 ( Lys92Arg or Ser229Ala , Ser232Ala) interacted with ERF104 independently of flg22 treatment. Treatment with aminoethoxyvinylglycine or silver ions removed flg22 inhibition of the interaction. The authors concluded that inhibition of interaction between MPK6 and ERF104 by flg22 required ethylene bio-synthesis and signalling. mpk6-erf104-1 830244 n/a 830247 enzyme target enzyme target substrate substrate neutral component neutral component region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Q9FKG1:p.[Ser229Ala;Ser232Ala] Alters kinetics of interaction 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 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) n/a 830248 enzyme enzyme neutral component neutral component arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress phosphorylation phosphorylation reaction phosphorylation false false false MPK6 was immunoprecipitated from flg22 peptide (QRLSTGSRINSAKDDAAGLQIA) treated Arabidopsis plants. Figure 3C Figure 3E Phosphorylation of ERF104 (Q9FKG1) mpk6-erf104-2 830245 n/a 830247 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 830248 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false SFigure 1D flg22 erf104-pdf1 830246 n/a 830247 unspecified role unspecified role bait bait region Strep tag strep tag strep tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 830249 unspecified role unspecified role prey prey promoter_region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false association association false false false Figure 5A