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/ miao-2007-2 Arabidopsis MEKK1 can take a short cut: it can directly interact with senescence-related WRKY53 transcription factor on the protein level and can bind to its promoter 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 Miao Y., Laun TM., Smykowski A., Zentgraf U. ulrike.zentgraf@uni-tuebingen.de Arabidopsis 7 week old seedling cDNA library Plant Mol. Biol. (0167-4412) 2007 miao-2007-1 Arabidopsis MEKK1 can take a short cut: it can directly interact with senescence-related WRKY53 transcription factor on the protein level and can bind to its promoter yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 1 hybrid one hybrid yeast one-hybrid yeast one hybrid one-hybrid 1 hybrid yeast one-hybrid one-hybrid yeast one hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Miao Y., Laun TM., Smykowski A., Zentgraf U. ulrike.zentgraf@uni-tuebingen.de Arabidopsis 7 week old seedling cDNA library Plant Mol. Biol. (0167-4412) 2007 miao-2007-3 Arabidopsis MEKK1 can take a short cut: it can directly interact with senescence-related WRKY53 transcription factor on the protein level and can bind to its promoter in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa predetermined predetermined participant predetermined Miao Y., Laun TM., Smykowski A., Zentgraf U. ulrike.zentgraf@uni-tuebingen.de Arabidopsis 7 week old seedling cDNA library Plant Mol. Biol. (0167-4412) 2007 miao-2007-4 Arabidopsis MEKK1 can take a short cut: it can directly interact with senescence-related WRKY53 transcription factor on the protein level and can bind to its promoter in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Miao Y., Laun TM., Smykowski A., Zentgraf U. ulrike.zentgraf@uni-tuebingen.de Arabidopsis 7 week old seedling cDNA library Plant Mol. Biol. (0167-4412) 2007 miao-2007-5 Arabidopsis MEKK1 can take a short cut: it can directly interact with senescence-related WRKY53 transcription factor on the protein level and can bind to its promoter arath-leaf Arabidopsis thaliana leaf leaf leaf bifc bimolecular fluorescence complementation bifc predetermined predetermined participant predetermined Miao Y., Laun TM., Smykowski A., Zentgraf U. ulrike.zentgraf@uni-tuebingen.de Arabidopsis 7 week old seedling cDNA library Plant Mol. Biol. (0167-4412) 2007 miao-2007-6 Arabidopsis MEKK1 can take a short cut: it can directly interact with senescence-related WRKY53 transcription factor on the protein level and can bind to its promoter arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress bifc bimolecular fluorescence complementation bifc predetermined predetermined participant predetermined Miao Y., Laun TM., Smykowski A., Zentgraf U. This study used Arabidopsis protoplasts ulrike.zentgraf@uni-tuebingen.de Arabidopsis 7 week old seedling cDNA library Plant Mol. Biol. (0167-4412) 2007 miao-2007-7 Arabidopsis MEKK1 can take a short cut: it can directly interact with senescence-related WRKY53 transcription factor on the protein level and can bind to its promoter in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Miao Y., Laun TM., Smykowski A., Zentgraf U. ulrike.zentgraf@uni-tuebingen.de Arabidopsis 7 week old seedling cDNA library Plant Mol. Biol. (0167-4412) 2007 miao-2007-8 Arabidopsis MEKK1 can take a short cut: it can directly interact with senescence-related WRKY53 transcription factor on the protein level and can bind to its promoter yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 1 hybrid one hybrid yeast one-hybrid yeast one hybrid one-hybrid 1 hybrid yeast one-hybrid one-hybrid yeast one hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Miao Y., Laun TM., Smykowski A., Zentgraf U. ulrike.zentgraf@uni-tuebingen.de Arabidopsis 7 week old seedling cDNA library Plant Mol. Biol. (0167-4412) 2007 wrk53_arath Probable WRKY transcription factor 53 WRKY53 At4g23810 F9D16.280 WRKY DNA-binding protein 53 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEGRDMLSWEQKTLLSELINGFDAAKKLQARLREAPSPSSSFSSPATAVAETNEILVKQIVSSYERSLLLLNWSSSPSVQLIPTPVTVVPVANPGSVPESPASINGSPRSEEFADGGGSSESHHRQDYIFNSKKRKMLPKWSEKVRISPERGLEGPQDDVFSWRKYGQKDILGAKFPRSYYRCTHRSTQNCWATKQVQRSDGDATVFEVTYRGTHTCSQAITRTPPLASPEKRQDTRVKPAITQKPKDILESLKSNLTVRTDGLDDGKDVFSFPDTPPFYNYGTINGEFGHVESSPIFDVVDWFNPTVEIDTTFPAFLHESIYY m3k1_arath Mitogen-activated protein kinase kinase kinase 1 MEKK1 At4g08500 T15F16.5 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDRILARMKKSTGRRGGDKNITPVRRLERRDAARNINYDAASCSSSSAEDLSVSTSSLMTRSLEFPEPTSFRIGGGVGEMDRIYRSLGVSGPDDLAISFDAWEACKKRSSSDVVNRFKSFDLDKVRDQDLSEEGPSGVVVGSDSMNHKVQGQDLSEAGPSGGIVTELSEIGNLITPVDRLVADGVVENRRVMERTPTIVKSKGYLVPNNVVAVGVGVGGGIKGLRPPVLKPPPAMKRPPIDHRGSSWDFLTHFAPSETVKRPSSSSSSSEDGCDEEEGKEEEAEAEEMGARFIQLGDTADETCSFTTNEGDSSSTVSNTSPIYPDGGAIITSWQKGQLLGRGSFGSVYEGISGDGDFFAVKEVSLLDQGSQAQECIQQLEGEIKLLSQLQHQNIVRYRGTAKDGSNLYIFLELVTQGSLLKLYQRYQLRDSVVSLYTRQILDGLKYLHDKGFIHRDIKCANILVDANGAVKLADFGLAKVSKFNDIKSCKGTPFWMAPEVINRKDSDGYGSPADIWSLGCTVLEMCTGQIPYSDLEPVQALFRIGRGTLPEVPDTLSLDARLFILKCLKVNPEERPTAAELLNHPFVRRPLPSVGSGGSGSASPLLRR wrky53-arath-dna WRKY53 promoter sequence -926 to -597 dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress WRKY53 promoter sequence -926 to -597 wrky53-mekk1-4 The MEKK1 protein binds the WRKY53 protein 671661 N/A 671669 unspecified role unspecified role bait bait region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false N/A 671670 unspecified role unspecified role prey prey physical association physical association false false false 6 mekk1-wrky53 The WRKY53 protein binds the MEKK1 protein 671661 N/A 671669 unspecified role unspecified role prey prey region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false N/A 671670 unspecified role unspecified role bait bait physical association physical association false false false 6 wrky53-mekk1 The MEKK1 protein binds a region of the WRKY53 promoter 671662 N/A 671671 unspecified role unspecified role bait bait N/A 671670 unspecified role unspecified role prey prey direct interaction direct interaction false false false 2 wrky53-mekk1-2 The MEKK1 protein binds a region of the WRKY53 promoter 671663 N/A 671670 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) N/A 671671 unspecified role unspecified role bait bait direct interaction direct interaction false false false 1B 2B 8B MEKK1 (Q39008) protein bound the CATGTG motif in the wild type WRKY53 promoter , the authors demonstrated that alteration of this motif to CTGTTG increased binding of MEKK1 but AATGTG or CATGTC motifs did not bind MEKK1. It was demonstrated that phosphorylation of WRKY53 protein positively influences the DNA-protein complex formation wrky53-mekk1-3 The MEKK1 protein binds a region of the WRKY53 promoter 671664 N/A 671671 unspecified role unspecified role bait bait N/A 671670 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) physical association physical association false false false 1C The nucleic acid was radio-labelled wrky53-mekk1-5 The MEKK1 protein binds the WRKY53 protein 671665 N/A 671669 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n-terminal N terminal region of GFP protein gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 671670 unspecified role unspecified role 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 c-terminal C terminal region of the GFP protein gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag 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 7 wrky53-mekk1-6 The MEKK1 protein binds the WRKY53 protein 671666 N/A 671669 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n-terminal N terminal region of GFP protein gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 671670 unspecified role unspecified role 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 c-terminal C terminal region of the GFP protein gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag 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 7 wrky53-mekk1-7 The MEKK1 protein phosphorylates the WRKY53 promoter 671667 N/A 671670 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) N/A 671669 enzyme target enzyme target substrate substrate neutral component neutral component ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false 8A wrky53-mekk1-1 The MEKK1 protein binds a region of the WRKY53 promoter 671668 N/A 671671 unspecified role unspecified role bait bait N/A 671670 unspecified role unspecified role prey prey direct interaction direct interaction false false false 1A The promoter sequence was from -926bp to -597bp from the transcription start site.