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/ The ZIM domain mediates homo- and heteromeric interactions between Arabidopsis JAZ proteins. The ZIM domain mediates homo- and heteromeric interactions between Arabidopsis JAZ proteins. Plant J. (0960-7412) 2009 imex curation Chini A., Fonseca S., Chico JM., Fernandez-Calvo P., Solano R. rsolano@cnb.csic.es 21-Dec-2010: Contacted by JYOTI. orchard Only protein-protein interactions nicbe Nicotiana benthamiana bifc bimolecular fluorescence complementation bifc tag fluorescence tag visualisation by fluorescence tag fluorescence Chini A., Fonseca S., Chico JM., Fernandez-Calvo P., Solano R. rsolano@cnb.csic.es Accepted 2009-JUL-8 by ORCHARD. 21-Dec-2010: Contacted by JYOTI. Plant J. (0960-7412) 2009 orchard Only protein-protein interactions imex curation The ZIM domain mediates homo- and heteromeric interactions between Arabidopsis JAZ proteins. The ZIM domain mediates homo- and heteromeric interactions between Arabidopsis JAZ proteins. Plant J. (0960-7412) 2009 imex curation Chini A., Fonseca S., Chico JM., Fernandez-Calvo P., Solano R. rsolano@cnb.csic.es 21-Dec-2010: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Chini A., Fonseca S., Chico JM., Fernandez-Calvo P., Solano R. rsolano@cnb.csic.es Accepted 2009-JUL-8 by ORCHARD. 21-Dec-2010: Contacted by JYOTI. Plant J. (0960-7412) 2009 orchard Only protein-protein interactions imex curation The ZIM domain mediates homo- and heteromeric interactions between Arabidopsis JAZ proteins. The ZIM domain mediates homo- and heteromeric interactions between Arabidopsis JAZ proteins. Plant J. (0960-7412) 2009 imex curation Chini A., Fonseca S., Chico JM., Fernandez-Calvo P., Solano R. rsolano@cnb.csic.es 21-Dec-2010: Contacted by JYOTI. orchard 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 Chini A., Fonseca S., Chico JM., Fernandez-Calvo P., Solano R. rsolano@cnb.csic.es Accepted 2009-JUL-8 by ORCHARD. 21-Dec-2010: Contacted by JYOTI. Plant J. (0960-7412) 2009 orchard Only protein-protein interactions imex curation The ZIM domain mediates homo- and heteromeric interactions between Arabidopsis JAZ proteins. The ZIM domain mediates homo- and heteromeric interactions between Arabidopsis JAZ proteins. Plant J. (0960-7412) 2009 imex curation Chini A., Fonseca S., Chico JM., Fernandez-Calvo P., Solano R. rsolano@cnb.csic.es 21-Dec-2010: Contacted by JYOTI. orchard 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 Chini A., Fonseca S., Chico JM., Fernandez-Calvo P., Solano R. rsolano@cnb.csic.es Accepted 2009-JUL-8 by ORCHARD. Plant J. (0960-7412) 2009 orchard Only protein-protein interactions imex curation ti11a_arath Protein TIFY 11A TIFY11A JAZ5 Jasmonate ZIM domain-containing protein 5 F1L3.3 F28G4.16 At1g17380 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSSSNENAKAQAPEKSDFTRRCSLLSRYLKEKGSFGNIDLGLYRKPDSSLALPGKFDPPGKQNAMHKAGHSKGEPSTSSGGKVKDVADLSESQPGSSQLTIFFGGKVLVYNEFPVDKAKEIMEVAKQAKPVTEINIQTPINDENNNNKSSMVLPDLNEPTDNNHLTKEQQQQQEQNQIVERIARRASLHRFFAKRKDRAVARAPYQVNQNAGHHRYPPKPEIVTGQPLEAGQSSQRPPDNAIGQTMAHIKSDGDKDDIMKIEEGQSSKDLDLRL DIP protein Q9LDU5 original sequence version: 50 DIP protein Q9LDU5 original sequence version: 78 DIP protein Q9LDU5 original sequence version: 87 54A55E02C18E5BAF ti10b_arath Protein TIFY 10B TIFY10B JAZ2 F9E10.20 F25A4.8 At1g74950 Jasmonate ZIM domain-containing protein 2 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSSFSAECWDFSGRKPSFSQTCTRLSRYLKEKGSFGDLSLGMTCKPDVNGGSRQPTMMNLFPCEASGMDSSAGQEDIKPKTMFPRQSSFSSSSSSGTKEDVQMIKETTKSVKPESQSAPLTIFYGGRVMVFDDFSAEKAKEVIDLANKGSAKSFTCFTAEVNNNHSAYSQKEIASSPNPVCSPAKTAAQEPIQPNPASLACELPIARRASLHRFLEKRKDRITSKAPYQIDGSAEASSKPTNPAWLSSR DIP protein Q9S7M2 original sequence version: 51 DIP protein Q9S7M2 original sequence version: 81 DIP protein Q9S7M2 original sequence version: 90 7410B2A50505A543 ti10a_arath Protein TIFY 10A TIFY10A JAZ1 At1g19180 T29M8.5 Jasmonate ZIM domain-containing protein 1 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSSSMECSEFVGSRRFTGKKPSFSQTCSRLSQYLKENGSFGDLSLGMACKPDVNGTLGNSRQPTTTMSLFPCEASNMDSMVQDVKPTNLFPRQPSFSSSSSSLPKEDVLKMTQTTRSVKPESQTAPLTIFYAGQVIVFNDFSAEKAKEVINLASKGTANSLAKNQTDIRSNIATIANQVPHPRKTTTQEPIQSSPTPLTELPIARRASLHRFLEKRKDRVTSKAPYQLCDPAKASSNPQTTGNMSWLGLAAEI DIP protein Q9LMA8 original sequence version: 76 DIP protein Q9LMA8 original sequence version: 72 DIP protein Q9LMA8 original sequence version: 51 DIP protein Q9LMA8 original sequence version: 57 DIP protein Q9LMA8 original sequence version: 74 DIP protein Q9LMA8 original sequence version: 81 DIP protein Q9LMA8 original sequence version: 92 EF2DAAC0CEF2DBBB tif6a_arath Protein TIFY 6A TIFY6A JAZ4 Jasmonate ZIM domain-containing protein 4 T1N15.11 At1g48500 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MERDFLGLGSKLSPITVKEETNEDSAPSRGMMDWSFSSKVGSGPQFLSFGTSQQETRVNTVNDHLLSSAAMDQNQRTYFSSLQEDRVFPGSSQQDQTTITVSMSEPNYINSFINHQHLGGSPIMAPPVSVFPAPTTIRSSSKPLPPQLTIFYAGSVLVYQDIAPEKAQAIMLLAGNGPHAKPVSQPKPQKLVHHSLPTTDPPTMPPSFLPSISYIVSETRSSGSNGVTGLGPTKTKASLASTRNNQTAAFSMAPTVGLPQTRKASLARFLEKRKERVINVSPYYVDNKSSIDCRTLMSECVSCPPAHHLH DIP protein Q58G47 original sequence version: 31 DIP protein Q58G47 original sequence version: 58 DIP protein Q58G47 original sequence version: 68 83E08ACF3E387ADC coi1_arath Coronatine-insensitive protein 1 FBL2 At2g39940 COI1 COI-1 F-box/LRR-repeat protein 2 T28M21.10 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEDPDIKRCKLSCVATVDDVIEQVMTYITDPKDRDSASLVCRRWFKIDSETREHVTMALCYTATPDRLSRRFPNLRSLKLKGKPRAAMFNLIPENWGGYVTPWVTEISNNLRQLKSVHFRRMIVSDLDLDRLAKARADDLETLKLDKCSGFTTDGLLSIVTHCRKIKTLLMEESSFSEKDGKWLHELAQHNTSLEVLNFYMTEFAKISPKDLETIARNCRSLVSVKVGDFEILELVGFFKAAANLEEFCGGSLNEDIGMPEKYMNLVFPRKLCRLGLSYMGPNEMPILFPFAAQIRKLDLLYALLETEDHCTLIQKCPNLEVLETRNVIGDRGLEVLAQYCKQLKRLRIERGADEQGMEDEEGLVSQRGLIALAQGCQELEYMAVYVSDITNESLESIGTYLKNLCDFRLVLLDREERITDLPLDNGVRSLLIGCKKLRRFAFYLRQGGLTDLGLSYIGQYSPNVRWMLLGYVGESDEGLMEFSRGCPNLQKLEMRGCCFSERAIAAAVTKLPSLRYLWVQGYRASMTGQDLMQMARPYWNIELIPSRRVPEVNQQGEIREMEHPAHILAYYSLAGQRTDCPTTVRVLKEPI DIP protein O04197 original sequence version: 103 DIP protein O04197 original sequence version: 98 DIP protein O04197 original sequence version: 80 DIP protein O04197 original sequence version: 84 DIP protein O04197 original sequence version: 101 DIP protein O04197 original sequence version: 119 1DDCF04990144C06 ti11b_arath Protein TIFY 11B TIFY11B JAZ6 Jasmonate ZIM domain-containing protein 6 T10D10.8 At1g72450 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSTGQAPEKSNFSQRCSLLSRYLKEKGSFGNINMGLARKSDLELAGKFDLKGQQNVIKKVETSETRPFKLIQKFSIGEASTSTEDKAIYIDLSEPAKVAPESGNSQLTIFFGGKVMVFNEFPEDKAKEIMEVAKEANHVAVDSKNSQSHMNLDKSNVVIPDLNEPTSSGNNEDQETGQQHQVVERIARRASLHRFFAKRKDRAVARAPYQVNQHGSHLPPKPEMVAPSIKSGQSSQHIATPPKPKAHNHMPMEVDKKEGQSSKNLELKL DIP protein Q9C9E3 original sequence version: 44 DIP protein Q9C9E3 original sequence version: 70 DIP protein Q9C9E3 original sequence version: 78 E67821C53F6B138F tif5a_arath Protein TIFY 5A TIFY5A JAZ8 Jasmonate ZIM domain-containing protein 8 T2H7 At1g30135 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MKLQQNCDLELRLFPTSYDSDSSDTTSVVESTSSGNPQPNEESQRITIFYNGKMCFSSDVTHLQARSIISIASREMKTKSSSNGSDPPNKSTSFHHNQLPNPKASMKKSLQSFLQKRKIRIQATSPYHSRR DIP protein Q8LBM2 original sequence version: 35 DIP protein Q8LBM2 original sequence version: 62 DIP protein Q8LBM2 original sequence version: 71 4C4F6E24842E5B2B tif3a_arath Protein TIFY 3A TIFY3A JAZ11 Jasmonate ZIM domain-containing protein 11 T18D12.10 At3g43440 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAEVNGDFPVPSFADGTGSVSAGLDLLVERSIHEARSTEPDASTQLTIIFGGSCRVFNGVPAQKVQEIIRIAFAGKQTKNVTGINPALNRALSFSTVADLPIARRRSLQRFLEKRRDRSTKPDGSMILPSQLTIIFGGSFSVFDGIPAEKVQEILHIAAAAKATETINLTSINPALKRAISFSNASTVACVSTADVPIARRRSLQRFFEKRRHRFVHTKPYSATTSEADKNETSPIVT DIP protein Q9M246 original sequence version: 49 DIP protein Q9M246 original sequence version: 85 841F602E9BE9EE9E tif7_arath Protein TIFY 7 TIFY7 JAZ9 F5A18.12 At1g70700 Jasmonate ZIM domain-containing protein 9 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MERDFLGLSDKQYLSNNVKHEVNDDAVEERGLSTKAAREWGKSKVFATSSFMPSSDFQEAKAFPGAYQWGSVSAANVFRRCQFGGAFQNATPLLLGGSVPLPTHPSLVPRVASSGSSPQLTIFYGGTISVFNDISPDKAQAIMLCAGNGLKGETGDSKPVREAERMYGKQIHNTAATSSSSATHTDNFSRCRDTPVAATNAMSMIESFNAAPRNMIPSVPQARKASLARFLEKRKERLMSAMPYKKMLLDLSTGESSGMNYSSTSPT DIP protein Q8W4J8 original sequence version: 38 DIP protein Q8W4J8 original sequence version: 58 DIP protein Q8W4J8-1 original sequence version: 59 DIP protein Q8W4J8 original sequence version: 68 DIP protein Q8W4J8-1 original sequence version: 78 855ACCF25C47B650 tif3b_arath Protein TIFY 3B TIFY3B JAZ12 Jasmonate ZIM domain-containing protein 12 F22D1.70 At5g20900 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MTKVKDEPRASVEGGCGVADGDGGAAEIGGTGSVEKSINEVRSTEIQTAEPTVPPNQLTIFFGGSVTVFDGLPSEKVQEILRIAAKAMETKNSTSISPVSSPALNRAPSFSSTSNVASPAAQPFPIQPISFCRSTADLPIARRHSLQRFLEKRRDRLVNKNPYPTSDFKKTDVPTGNVSIKEEFPTA DIP protein Q9C5K8 original sequence version: 48 DIP protein Q9C5K8 original sequence version: 86 4F0C66DECBB4F0E1 myc2_arath Transcription factor MYC2 MYC2 BHLH6 Basic helix-loop-helix protein 6 EN38 JAI1 JIN1 Protein JASMONATE INSENSITIVE 1 R-homologous Arabidopsis protein 1 RAP1 RD22BP1 Transcription factor EN 38 Z-box binding factor 1 protein ZBF1 bHLH transcription factor bHLH006 rd22BP1 F6N18.4 At1g32640 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MTDYRLQPTMNLWTTDDNASMMEAFMSSSDISTLWPPASTTTTTATTETTPTPAMEIPAQAGFNQETLQQRLQALIEGTHEGWTYAIFWQPSYDFSGASVLGWGDGYYKGEEDKANPRRRSSSPPFSTPADQEYRKKVLRELNSLISGGVAPSDDAVDEEVTDTEWFFLVSMTQSFACGAGLAGKAFATGNAVWVSGSDQLSGSGCERAKQGGVFGMHTIACIPSANGVVEVGSTEPIRQSSDLINKVRILFNFDGGAGDLSGLNWNLDPDQGENDPSMWINDPIGTPGSNEPGNGAPSSSSQLFSKSIQFENGSSSTITENPNLDPTPSPVHSQTQNPKFNNTFSRELNFSTSSSTLVKPRSGEILNFGDEGKRSSGNPDPSSYSGQTQFENKRKRSMVLNEDKVLSFGDKTAGESDHSDLEASVVKEVAVEKRPKKRGRKPANGREEPLNHVEAERQRREKLNQRFYALRAVVPNVSKMDKASLLGDAIAYINELKSKVVKTESEKLQIKNQLEEVKLELAGRKASASGGDMSSSCSSIKPVGMEIEVKIIGWDAMIRVESSKRNHPAARLMSALMDLELEVNHASMSVVNDLMIQQATVKMGFRIYTQEQLRASLISKIG DIP protein Q39204 original sequence version: 109 DIP protein Q39204 original sequence version: 104 DIP protein Q39204 original sequence version: 107 DIP protein Q39204 original sequence version: 126 4A8D4DE34C5928F6 tif9_arath Protein TIFY 9 TIFY9 JAS1 JAZ10 Protein JASMONATE-ASSOCIATED 1 Jasmonate ZIM domain-containing protein 10 Protein JAZ10 T31B5.40 At5g13220 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSKATIELDFLGLEKKQTNNAPKPKFQKFLDRRRSFRDIQGAISKIDPEIIKSLLASTGNNSDSSAKSRSVPSTPREDQPQIPISPVHASLARSSTELVSGTVPMTIFYNGSVSVFQVSRNKAGEIMKVANEAASKKDESSMETDLSVILPTTLRPKLFGQNLEGDLPIARRKSLQRFLEKRKERLVSTSPYYPTSA DIP protein Q93ZM9 original sequence version: 44 DIP protein Q93ZM9 original sequence version: 70 DIP protein Q93ZM9 original sequence version: 79 475DADBCEDC540E1 tif6b_arath Protein TIFY 6B TIFY6B JAI3 JAZ3 MEB5.8 At3g17860 Jasmonate ZIM domain-containing protein 3 Protein JASMONATE INSENSITIVE 3 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MERDFLGLGSKNSPITVKEETSESSRDSAPNRGMNWSFSNKVSASSSQFLSFRPTQEDRHRKSGNYHLPHSGSFMPSSVADVYDSTRKAPYSSVQGVRMFPNSNQHEETNAVSMSMPGFQSHHYAPGGRSFMNNNNNSQPLVGVPIMAPPISILPPPGSIVGTTDIRSSSKPIGSPAQLTIFYAGSVCVYDDISPEKAKAIMLLAGNGSSMPQVFSPPQTHQQVVHHTRASVDSSAMPPSFMPTISYLSPEAGSSTNGLGATKATRGLTSTYHNNQANGSNINCPVPVSCSTNVMAPTVALPLARKASLARFLEKRKERVTSVSPYCLDKKSSTDCRRSMSECISSSLSSAT DIP protein Q9LVI4 original sequence version: 75 DIP protein Q9LVI4 original sequence version: 51 DIP protein Q9LVI4 original sequence version: 72 DIP protein Q9LVI4 original sequence version: 81 DIP protein Q9LVI4 original sequence version: 91 D65DEEA2F04C58E4 col1-tify6b-2 2 17 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 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 arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress physical association physical association Figure 1a Interaction of COI1-flag with MBP-JAZ3 and truncated versions. Immunoblot (anti-flag antibody) of recovered COI1-flag 50 micro molar coronatine col1-tify6b-3 4 17 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 9 unspecified role unspecified role prey prey physical association physical association Figure 1b As shown in Figure 1a, full-length MBP�JAZ3 and MBP�JAZ3ΔZIM proteins, but not MBP�JAZ3ΔJas, were able to pull-down COI1-flag from the extracts in the presence of COR (a hormone agonist), indicating that the Jas motif is required for the hormone-dependent interaction between JAZ3 and COI1. Yeast two-hybrid analysis consistently confirmed these results (Figure 1b). 50 micro molar coronatine tify6b-myc2 2 17 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 2c Interaction between MYC2-GFP and MBP-JAZ3 full-length or truncated protein versions. Immunoblot (anti-GFP antibody) of recovered MYC2-GFP (from 35S:MYC2-GFP;jin1-2 plant extracts) after pull-down reactions using recombinant MBP-JAZ3 protein and truncated versions tify6b-myc2-1 4 17 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 15 unspecified role unspecified role prey prey physical association physical association Figure 2d To further define a shorter domain in the JAZ proteins required for MYC2 interaction, we performed yeast two-hybrid assays with additional JAZ3 derivatives. As shown in Figure 2d, in addition to JAZ3 and JAZ3ΔZIM proteins (previously shown to interact with MYC2; Chini et al., 2007), the Jas motif alone (Jas) was sufficient for interaction tify10a-myc2-1 2 7 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 3a The authors concluded that the interaction was strong in the case of JAZ1. tify10b-myc2 2 6 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 15 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 3a The authors concluded that the interaction was strong in the case of JAZ2. tify6a-myc2-1 2 8 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 3a The authors concluded that the interaction was weak in the case of JAZ4. tify11a-myc2-2 2 5 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 3a As shown in Figure 3a, MYC2-GFP was recovered by all MBP-JAZ proteins, except for JAZ7, in PD experiments. The interaction was strong in the case of JAZ1, JAZ2, JAZ3, JAZ5, JAZ9, JAZ10 and JAZ11, and was weak in the case of JAZ4, JAZ6, JAZ8 and JAZ12. Consistently, most JAZ proteins, with the exceptions of JAZ4 and JAZ7, interacted with MYC2 in yeast two-hybrid assays, also with different affinitie (Figure 3b). We conclude that most JAZ proteins can interact with MYC2. The authors concluded that the interaction was strong in the case of JAZ5. tify11b-myc2-3 2 10 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 3a The authors concluded that the interaction was weak in the case of JAZ6. tify5a-myc2-4 2 11 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 3a The authors concluded that the interaction was weak in the case of JAZ8. tify7-myc2-1 2 13 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 3a The authors concluded that the interaction was strong in the case of JAZ9. tify9-myc2 2 16 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 3a The authors concluded that the interaction was strong in the case of JAZ10. tify3a-myc2 2 12 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 3a The authors concluded that the interaction was strong in the case of JAZ11. tify3b-myc2 2 14 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 3a The authors concluded that the interaction was weak in the case of JAZ12. tify10a-myc2-2 4 7 unspecified role unspecified role bait bait 15 unspecified role unspecified role prey prey physical association physical association Figure 3b Figure 3c tify10b-myc2-1 4 6 unspecified role unspecified role bait bait 15 unspecified role unspecified role prey prey physical association physical association Figure 3b Figure 3c tify11a-myc2-3 4 5 unspecified role unspecified role bait bait 15 unspecified role unspecified role prey prey physical association physical association Figure 3b Figure 3c tify5a-myc2-5 4 11 unspecified role unspecified role bait bait 15 unspecified role unspecified role prey prey physical association physical association Figure 3b Figure 3c tify7-myc2-2 4 13 unspecified role unspecified role bait bait 15 unspecified role unspecified role prey prey physical association physical association Figure 3b Figure 3c tify9-myc2-1 4 16 unspecified role unspecified role bait bait 15 unspecified role unspecified role prey prey physical association physical association Figure 3b Figure 3c tify3a-myc2-1 2 12 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 3a The authors concluded that the interaction was strong in the case of JAZ11. tify3b-myc2-1 4 14 unspecified role unspecified role bait bait 15 unspecified role unspecified role prey prey physical association physical association Figure 3b Figure 3c tify11b-myc2-4 4 10 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role prey prey physical association physical association Figure 3c tify10a-1 4 7 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey physical association physical association Figure 5 Figure 6 tify6b-2 4 17 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 17 unspecified role unspecified role prey prey physical association physical association Figure 4d Figure 5a tify6a-2 4 8 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association Figure 5a tify7-3 4 13 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait physical association physical association Figure 5a Figure 6 tify6b-3 2 17 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress 17 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Figure 4c To identify the domain responsible for dimerization, we used JAZ3 derivatives in PD experiments, defining the ZIM domain as the motif necessary and sufficient for homomeric interactions tify10a-tify6a 4 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Figure 6 tify10a-tify5a 4 7 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey physical association physical association Figure 6 tify10a-tify7 4 7 unspecified role unspecified role bait bait 13 unspecified role unspecified role prey prey physical association physical association Figure 6 tify6a-tify6b 4 8 unspecified role unspecified role bait bait 17 unspecified role unspecified role prey prey 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 Figure 6 Figure 7 tify6b-tify6a 4 17 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association Figure 6 tify6b-tify7 4 17 unspecified role unspecified role bait bait 13 unspecified role unspecified role prey prey physical association physical association Figure 6 tify6a-tify7 4 13 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association Figure 6 tiry5a-tify10a-1 4 7 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association Figure 6 tify7-tify6b 4 17 unspecified role unspecified role prey prey n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 13 unspecified role unspecified role bait bait physical association physical association Figure 6 Figure 7a tify7-tify6a 4 8 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait physical association physical association Figure 6 tify10a-tify6b 4 7 unspecified role unspecified role bait bait 17 unspecified role unspecified role prey prey 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 Figure 7a tify6b-1 2 17 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 17 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress physical association physical association Figure 7b Consistent with the yeast two-hybrid results, JAZ1, JAZ3, JAZ4 and JAZ9 were the proteins that most strongly pulled-down JAZ3ΔJas�GFP from the extracts. JAZ2, JAZ5, JAZ6 and JAZ10 only showed a weak interaction with JAZ3ΔJas�GFP, and no interaction over the background was observed for JAZ7, JAZ8, JAZ11 and JAZ12. These results further support the existence of heteromeric interactions among JAZ proteins involving the ZIM domain. The authors concluded the bait and prey interaction was 'strong'. tify6a-tify10a 2 7 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 17 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress physical association physical association Figure 7b The authors concluded the bait and prey interaction was 'strong'. tify6a-tify6b-1 2 8 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 17 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress physical association physical association Figure 7b The authors concluded the bait and prey interaction was 'strong'. tify6a-tify10b 2 6 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 17 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress physical association physical association Figure 7b The authors concluded the bait and prey interaction was 'weak'. tify6a-tify11a 2 5 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 17 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress physical association physical association Figure 7b The authors concluded the bait and prey interaction was 'weak'. tify6a-tify11b 2 17 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress 10 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Figure 7b The authors concluded the bait and prey interaction was 'weak'. tify6a-tify9 2 17 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress 16 unspecified role unspecified role bait bait n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Figure 7b The authors concluded the bait and prey interaction was 'weak'. tify6b-4 1 17 unspecified role unspecified role neutral component neutral component region N-terminal YFP 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 17 unspecified role unspecified role neutral component neutral component n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region C-terminal YFP 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 Figure 8 As shown in Figure 8a, fusion proteins of JAZ3ΔJas to the N-terminal and C-terminal halves of YFP (JAZ3ΔJas-NY and CY-JAZ3ΔJas, respectively) complemented the YFP, and produced fluorescent nuclei at the YFP wavelength tify6b-5 3 17 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain nicbe Nicotiana benthamiana 17 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 ecoli Escherichia coli (strain K12) physical association physical association Figure 8b tify6b-tify7-1 3 17 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 ecoli Escherichia coli (strain K12) 13 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined nicbe Nicotiana benthamiana physical association physical association Figure 8b