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/ Direct interaction between the Arabidopsis disease resistance signaling proteins, EDS1 and PAD4 Direct interaction between the Arabidopsis disease resistance signaling proteins, EDS1 and PAD4 EMBO J. (0261-4189) 2001 Feys BJ., Moisan LJ., Newman MA., Parker JE. parker@mpiz-koeln.mpg.de 13-Nov-2007: Contacted by JYOTI. orchard 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 Feys BJ., Moisan LJ., Newman MA., Parker JE. parker@mpiz-koeln.mpg.de Accepted 2007-NOV-19 by ORCHARD. EMBO J. (0261-4189) 2001 orchard Direct interaction between the Arabidopsis disease resistance signaling proteins, EDS1 and PAD4 Direct interaction between the Arabidopsis disease resistance signaling proteins, EDS1 and PAD4 EMBO J. (0261-4189) 2001 Feys BJ., Moisan LJ., Newman MA., Parker JE. parker@mpiz-koeln.mpg.de 13-Nov-2007: Contacted by JYOTI. orchard 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 Feys BJ., Moisan LJ., Newman MA., Parker JE. parker@mpiz-koeln.mpg.de Accepted 2007-NOV-19 by ORCHARD. 13-Nov-2007: Contacted by JYOTI. Arabidopsis cDNA library made from Peronospora parasitica pathogen challenged plants. EMBO J. (0261-4189) 2001 orchard Direct interaction between the Arabidopsis disease resistance signaling proteins, EDS1 and PAD4 Direct interaction between the Arabidopsis disease resistance signaling proteins, EDS1 and PAD4 EMBO J. (0261-4189) 2001 Feys BJ., Moisan LJ., Newman MA., Parker JE. parker@mpiz-koeln.mpg.de 13-Nov-2007: Contacted by JYOTI. orchard arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Feys BJ., Moisan LJ., Newman MA., Parker JE. A null mutant arabidopsis pad4 line was stably transformed with wild-type PAD4 containing an N-terminal c-Myc epitope tag expressed from the native PAD4 promoter (Myc::PAD4). parker@mpiz-koeln.mpg.de Accepted 2007-NOV-19 by ORCHARD. 13-Nov-2007: Contacted by JYOTI. EMBO J. (0261-4189) 2001 orchard eds1c_arath Protein EDS1 EDS1 EDS1-90 EDS1A Enhanced disease susceptibility 1 T17F15.40 At3g48090 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAFEALTGINGDLITRSWSASKQAYLTERYHKEEAGAVVIFAFQPSFSEKDFFDPDNKSSFGEIKLNRVQFPCMRKIGKGDVATVNEAFLKNLEAIIDPRTSFQASVEMAVRSRKQIVFTGHSSGGATAILATVWYLEKYFIRNPNVYLEPRCVTFGAPLVGDSIFSHALGREKWSRFFVNFVSRFDIVPRIMLARKASVEETLPHVLAQLDPRKSSVQESEQRITEFYTRVMRDTSTVANQAVCELTGSAEAFLETLSSFLELSPYRPAGTFVFSTEKRLVAVNNSDAILQMLFYTSQASDEQEWSLIPFRSIRDHHSYEELVQSMGKKLFNHLDGENSIESTLNDLGVSTRGRQYVQAALEEEKKRVENQKKIIQVIEQERFLKKLAWIEDEYKPKCQAHKNGYYDSFKVSNEENDFKANVKRAELAGVFDEVLGLMKKCQLPDEFEGDIDWIKLATRYRRLVEPLDIANYHRHLKNEDTGPYMKRGRPTRYIYAQRGYEHYILKPNGMIAEDVFWNKVNGLNLGLQLEEIQETLKNSGSECGSCFWAEVEELKGKPYEEVEVRVKTLEGMLGEWITDGEVDDKEIFLEGSTFRKWWITLPKNHKSHSPLRDYMMDEITDT F9B321D5DF77D96B pad4_arath Lipase-like PAD4 PAD4 EDS9 Protein ENHANCED DISEASE SUSCEPTIBILITY 9 Protein PHYTOALEXIN DEFICIENT 4 F22O6.190 At3g52430 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDDCRFETSELQASVMISTPLFTDSWSSCNTANCNGSIKIHDIAGITYVAIPAVSMIQLGNLVGLPVTGDVLFPGLSSDEPLPMVDAAILKLFLQLKIKEGLELELLGKKLVVITGHSTGGALAAFTALWLLSQSSPPSFRVFCITFGSPLLGNQSLSTSISRSRLAHNFCHVVSIHDLVPRSSNEQFWPFGTYLFCSDKGGVCLDNAGSVRLMFNILNTTATQNTEEHQRYGHYVFTLSHMFLKSRSFLGGSIPDNSYQAGVALAVEALGFSNDDTSGVLVKECIETATRIVRAPILRSAELANELASVLPARLEIQWYKDRCDASEEQLGYYDFFKRYSLKRDFKVNMSRIRLAKFWDTVIKMVETNELPFDFHLGKKWIYASQFYQLLAEPLDIANFYKNRDIKTGGHYLEGNRPKRYEVIDKWQKGVKVPEECVRSRYASTTQDTCFWAKLEQAKEWLDEARKESSDPQRRSLLREKIVPFESYANTLVTKKEVSLDVKAKNSSYSVWEANLKEFKCKMGYENEIEMVVDESDAMET D6D5D3EBB522C11A pad4-eds1 1 4 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 2 We found that PAD4 interacted with full-length EDS1 but not with any EDS1 subdomain tested, as shown in Figure 2B. PAD4 association with EDS1 was through its N-terminal region, comprising the predicted lipase domain. eds1-eds1 1 4 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait Q9SU72:p.Glu466Lys mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain E K Wed May 02 19:33:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu466lys' Tue Jun 30 19:10:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu466Lys' c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 2 The C-terminus of EDS1 was sufficient for interaction with full-length EDS1 (Figure 2B), suggesting that EDS1 dimerization occurs through the C-terminal end. eds1-pad4 2 5 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait physical association physical association fter transformation of the two-hybrid library in yeast strain EGY191(p8op- LacZ/pLex-EDS1), 6 000 000 primary transformants were obtained and 60 000 000 yeast clones were screened for potential EDS1 interactors. The dominant class of interactor (in 11 isolates) was identified as PAD4. Only full-length PAD4 inserts were recovered from the screen. EDS1�PAD4 interaction was also tested in the reciprocal combination with PAD4 fused to the LexA domain and was found to be stronger than the original interaction (data not shown). EDS1 pad4-eds1-1 2 5 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey physical association physical association fter transformation of the two-hybrid library in yeast strain EGY191(p8op- LacZ/pLex-EDS1), 6 000 000 primary transformants were obtained and 60 000 000 yeast clones were screened for potential EDS1 interactors. The dominant class of interactor (in 11 isolates) was identified as PAD4. Only full-length PAD4 inserts were recovered from the screen. EDS1�PAD4 interaction was also tested in the reciprocal combination with PAD4 fused to the LexA domain and was found to be stronger than the original interaction (data not shown). EDS1 eds1-pad4-1 3 4 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey n-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Immunoprecipitation of EDS1 protein from extracts of the Myc::PAD4 line using the anti-EDS1 antiserum, followed by detection on western blots using the anti-c-Myc antibody, showed that the EDS1 and PAD4 proteins interact in healthy plant tissue 4 The authors state that after 6 days of exposure to a virulent P.parasitica isolate, Emwa1, enhanced levels of both proteins correlated with increased detection of co-immunoprecipitable PAD4. Also in cases where leaves were sprayed with the bioactive SA analogue, benzothiadiazole (BTH), elevated EDS1 and PAD4 expression corresponded with higher levels of co-immunoprecipitable PAD4 24 h after treatment. Finally in leaves of mature plants were infiltrated with the virulent P.syringae pv. tomato strain, DC3000, or with avirulent DC3000 expressing avrRps4 (DC3000/avrRps4), enhanced levels of EDS1 and PAD4 protein were observed in tissues responding to DC3000/avrRps4 12 h after infiltration and these persisted for up to 48 h 6 days of exposure to a virulent P.parasitica isolate 24 hours after exposure to benzothiadiazole (BTH) 12 hours exposure to virulent P.syringae pv. tomato strain, DC3000 12 hours exposure to avirulent DC3000 expressing avrRps4 (DC3000/avrRps4)