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/ Arabidopsis WRKY38 and WRKY62 Transcription Factors Interact with Histone Deacetylase 19 in Basal Defense. Arabidopsis WRKY38 and WRKY62 Transcription Factors Interact with Histone Deacetylase 19 in Basal Defense. Plant Cell (1040-4651) 2008 imex curation Kim KC., Lai Z., Fan B., Chen Z. zhixiang@purdue.edu 16-Oct-2008: Contacted by JYOTI. jyoti Only protein-protein interactions nicbe Nicotiana benthamiana bifc bimolecular fluorescence complementation bifc tag fluorescence tag visualisation by fluorescence tag fluorescence Kim KC., Lai Z., Fan B., Chen Z. zhixiang@purdue.edu Accepted 2008-OCT-22 by JYOTI. 16-Oct-2008: Contacted by JYOTI. Plant Cell (1040-4651) 2008 jyoti Only protein-protein interactions imex curation Arabidopsis WRKY38 and WRKY62 Transcription Factors Interact with Histone Deacetylase 19 in Basal Defense. Arabidopsis WRKY38 and WRKY62 Transcription Factors Interact with Histone Deacetylase 19 in Basal Defense. Plant Cell (1040-4651) 2008 imex curation Kim KC., Lai Z., Fan B., Chen Z. zhixiang@purdue.edu 16-Oct-2008: Contacted by JYOTI. jyoti Only protein-protein interactions 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 Kim KC., Lai Z., Fan B., Chen Z. The yeast CytoTrap system based on interactiondependent activation of the Ras signaling pathway and cell growth in a temperature-sensitive yeast mutant. zhixiang@purdue.edu Accepted 2008-OCT-22 by JYOTI. 16-Oct-2008: Contacted by JYOTI. Plant Cell (1040-4651) 2008 jyoti Only protein-protein interactions imex curation Arabidopsis WRKY38 and WRKY62 Transcription Factors Interact with Histone Deacetylase 19 in Basal Defense. Arabidopsis WRKY38 and WRKY62 Transcription Factors Interact with Histone Deacetylase 19 in Basal Defense. Plant Cell (1040-4651) 2008 imex curation Kim KC., Lai Z., Fan B., Chen Z. zhixiang@purdue.edu 16-Oct-2008: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Kim KC., Lai Z., Fan B., Chen Z. zhixiang@purdue.edu Accepted 2008-OCT-22 by JYOTI. 16-Oct-2008: Contacted by JYOTI. Plant Cell (1040-4651) 2008 jyoti Only protein-protein interactions imex curation Arabidopsis WRKY38 and WRKY62 Transcription Factors Interact with Histone Deacetylase 19 in Basal Defense. Arabidopsis WRKY38 and WRKY62 Transcription Factors Interact with Histone Deacetylase 19 in Basal Defense. Plant Cell (1040-4651) 2008 imex curation Kim KC., Lai Z., Fan B., Chen Z. zhixiang@purdue.edu 16-Oct-2008: Contacted by JYOTI. jyoti Only protein-protein interactions 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 Kim KC., Lai Z., Fan B., Chen Z. The yeast CytoTrap system based on interactiondependent activation of the Ras signaling pathway and cell growth in a temperature-sensitive yeast mutant. zhixiang@purdue.edu Accepted 2008-OCT-22 by JYOTI. 16-Oct-2008: Contacted by JYOTI. Arabidopsis plants sprayed with 2mM salicyclic acid 4 hours before harvesting and cDNA library construction. Plant Cell (1040-4651) 2008 jyoti Only protein-protein interactions imex curation Arabidopsis WRKY38 and WRKY62 Transcription Factors Interact with Histone Deacetylase 19 in Basal Defense. Arabidopsis WRKY38 and WRKY62 Transcription Factors Interact with Histone Deacetylase 19 in Basal Defense. Plant Cell (1040-4651) 2008 imex curation Kim KC., Lai Z., Fan B., Chen Z. zhixiang@purdue.edu 16-Oct-2008: Contacted by JYOTI. jyoti Only protein-protein interactions nicbe Nicotiana benthamiana anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Kim KC., Lai Z., Fan B., Chen Z. Transgenic Arabidopsis plants expressing epitope tagged proteins of interest were used in this study. zhixiang@purdue.edu Accepted 2008-OCT-22 by JYOTI. 16-Oct-2008: Contacted by JYOTI. Plant Cell (1040-4651) 2008 jyoti Only protein-protein interactions imex curation wbox-arath-dna ds dna double stranded deoxyribonucleic acid ds DNA ds dna arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress AACTTTGACCAATCTTTCAAGTA The oligonucleotide contains a W box (TTGACC/T). AC575D833A8F9C19 wrk18_arath WRKY transcription factor 18 WRKY18 WRKY DNA-binding protein 18 F28M20.10 At4g31800 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDGSSFLDISLDLNTNPFSAKLPKKEVSVLASTHLKRKWLEQDESASELREELNRVNSENKKLTEMLARVCESYNELHNHLEKLQSRQSPEIEQTDIPIKKRKQDPDEFLGFPIGLSSGKTENSSSNEDHHHHHQQHEQKNQLLSCKRPVTDSFNKAKVSTVYVPTETSDTSLTVKDGFQWRKYGQKVTRDNPSPRAYFRCSFAPSCPVKKKVQRSAEDPSLLVATYEGTHNHLGPNASEGDATSQGGSSTVTLDLVNGCHRLALEKNERDNTMQEVLIQQMASSLTKDSKFTAALAAAISGRLMEQSRT 762E6B520AC3FF8D wrk38_arath Probable WRKY transcription factor 38 WRKY38 WRKY DNA-binding protein 38 MQJ16.11 At5g22570 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEMNSPHEKAVQAIRYGHSCAMRLKRRLNHPMADGGPLSSYDLAKSIVESFSNAISILSAKPETEDDQFSDLSSRDSSPPPQGSPSKKRKIDSTNSSENWRDDSPDPIYYDGYLWRKYGQKSIKKSNHQRSYYRCSYNKDHNCEARKHEQKIKDNPPVYRTTYFGHHTCKTEHNLDAIFIAGQDPLDDFKSTQMIRFGKDQDQEKESRSNGFSLSVKHEEDIIKEQAIDQYREITSNDQDCQDVIEEYLSSPSGSYPPSSSSGSESADFNSDLLFDNPDSWDRYDQFYF 2C9CFA6F3E0D7026 wrk62_arath Probable WRKY transcription factor 62 WRKY62 WRKY DNA-binding protein 62 T20L15.170 At5g01900 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MNSCQQKAMEKLLHGHGCANQLLIMDQTESDSSMEREDLAKSVLHCFSDALSILIDTNDHQDDQSNNSSPQDSSPVLESSRKPLHKRGRKTSMAESSDYHRHESSTPIYHDGFLWRKYGQKQIKESEYQRSYYKCAYTKDQNCEAKKQVQKIQHNPPLYSTTYFGQHICQLHQAYATFPIDTSDFEEHEGSHMIRFGHPNISFSSSTSNLRQHQNHQDRIKDEYMKPVIAEDWSPSQWMSSEVALAVEAFEFNPFWTSHDLSS E8348EF30EE96995 hda19_arath Histone deacetylase 19 HDA19 At4g38130 F20D10.250 HD1 HDA1 RPD3A protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDTGGNSLASGPDGVKRKVCYFYDPEVGNYYYGQGHPMKPHRIRMTHALLAHYGLLQHMQVLKPFPARDRDLCRFHADDYVSFLRSITPETQQDQIRQLKRFNVGEDCPVFDGLYSFCQTYAGGSVGGSVKLNHGLCDIAINWAGGLHHAKKCEASGFCYVNDIVLAILELLKQHERVLYVDIDIHHGDGVEEAFYATDRVMTVSFHKFGDYFPGTGHIQDIGYGSGKYYSLNVPLDDGIDDESYHLLFKPIMGKVMEIFRPGAVVLQCGADSLSGDRLGCFNLSIKGHAECVKFMRSFNVPLLLLGGGGYTIRNVARCWCYETGVALGVEVEDKMPEHEYYEYFGPDYTLHVAPSNMENKNSRQMLEEIRNDLLHNLSKLQHAPSVPFQERPPDTETPEVDEDQEDGDKRWDPDSDMDVDDDRKPIPSRVKREAVEPDTKDKDGLKGIMERGKGCEVEVDESGSTKVTGVNPVGVEEASVKMEEEGTNKGGAEQAFPPKT DIP protein O22446 original sequence version: 96 857D2E3D16B7CC1F wrk40_arath Probable WRKY transcription factor 40 WRKY40 WRKY DNA-binding protein 40 F23A5.19 At1g80840 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDQYSSSLVDTSLDLTIGVTRMRVEEDPPTSALVEELNRVSAENKKLSEMLTLMCDNYNVLRKQLMEYVNKSNITERDQISPPKKRKSPAREDAFSCAVIGGVSESSSTDQDEYLCKKQREETVVKEKVSRVYYKTEASDTTLVVKDGYQWRKYGQKVTRDNPSPRAYFKCACAPSCSVKKKVQRSVEDQSVLVATYEGEHNHPMPSQIDSNNGLNRHISHGGSASTPVAANRRSSLTVPVTTVDMIESKKVTSPTSRIDFPQVQKLLVEQMASSLTKDPNFTAALAAAVTGKLYQQNHTEK F8AFC891237CBA1D wrky62-dna 3 6 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) direct interaction direct interaction Supplementary Figure 1B A protein/DNA complex with reduced mobility was detected when purified recombinant WRKY38 or WRKY62 protein was incubated with the P12a probe The authors noted that "binding of the WRKY proteins to a mutant probe (mP12a) in which the TTGACC sequence was changed to TTGAAC was not detectable". The authors conclude "binding of WRKY38 and WRKY62 to the TTGACC W-box sequence is highly specific". wrky38-dna 3 8 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role bait bait direct interaction direct interaction Supplementary Figure 1B A protein/DNA complex with reduced mobility was detected when purified recombinant WRKY38 or WRKY62 protein was incubated with the P12a probe The authors noted that "binding of the WRKY proteins to a mutant probe (mP12a) in which the TTGACC sequence was changed to TTGAAC was not detectable". The authors conclude "binding of WRKY38 and WRKY62 to the TTGACC W-box sequence is highly specific". wrky62-hda19 4 9 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association In these screens, we used a fusion protein of WRKY62 with the human homolog of yeast cdc25, hSOS, as bait. The temperature-sensitive yeast strain transformed with the bait construct failed to grow at the restrictive temperature (378C). We screened >107 independent transformants of a cDNA prey library generated from SA-treated Arabidopsis plants. The screens yielded one group of cDNAs that all encoded HDA19 wrky62-hda19-1 2 9 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association WRKY62 interaction with HDA19 in yeast. Yeast cdc25H cells containing the indicated plasmids were grown on synthetic minimal medium containing glucose or galactose at 25�C or 37�C. The pictures were taken after 4 days of growth. Supplementary Figure 4A wrky62-hda19-2 1 10 unspecified role unspecified role neutral component neutral component region C-terminal region YFP protein 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 9 unspecified role unspecified role neutral component neutral component region N-terminal region YFP protein 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 direct interaction direct interaction WhenWRKY62-N-YFP was coexpressed with HDA19-C-YFP, a strong BiFC signal was observed predominantly in the nuclear compartment, based on staining with 4,6-diamidino-2-phenylindole (DAPI) Figure 5 wrky38-hda19-3 1 8 unspecified role unspecified role neutral component neutral component region N-terminal region YFP protein 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 10 unspecified role unspecified role neutral component neutral component region C-terminal region YFP protein 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 direct interaction direct interaction Indeed, whenWRKY38 was fused to the terminal YFP fragment and coexpressed with HDA19-C-YFP in tobacco leaves, a BiFC signal was also observed predominantly in the nuclear compartment Figure 5 wrky18-wrky40 1 7 unspecified role unspecified role neutral component neutral component 11 unspecified role unspecified role neutral component neutral component direct interaction direct interaction Supplementary Figure 5 when WRKY40 was fused to the C-terminal YFP fragment and coexpressed with WRKY18-NYFP in tobacco leaves, a strong BiFC signal was observed predominantly in the nuclear compartment wrky38-hda19-4 5 10 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association The protein complexes immunoprecipitated by the anti-MYC antibody from protein extracts of leaves coexpressing MYC-HDA19 and FLAG-WRKY38 or MYC-HDA19 and FLAG-WRKY62 generated positive interactions to the anti-FLAG antisera Supplementary Figure 4B wrky62-hda19-4 5 10 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction The protein complexes immunoprecipitated by the anti-MYC antibody from protein extracts of leaves coexpressing MYC-HDA19 and FLAG-WRKY38 or MYC-HDA19 and FLAG-WRKY62 generated positive interactions to the anti-FLAG antisera Supplementary Figure 4B