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/ Modulation of drought resistance by the abscisic acid-receptor PYL5 through inhibition of clade A PP2Cs. Modulation of drought resistance by the abscisic acid-receptor PYL5 through inhibition of clade A PP2Cs. Plant J. (0960-7412) 2009 imex curation Santiago J., Rodrigues A., Saez A., Rubio S., Antoni R., Dupeux F., Park SY., Marquez JA., Cutler SR., Rodriguez PL. prodiguez@ibmcp.upv.es 27-Oct-2007: 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 Santiago J., Rodrigues A., Saez A., Rubio S., Antoni R., Dupeux F., Park SY., Marquez JA., Cutler SR., Rodriguez PL. prodiguez@ibmcp.upv.es Accepted 2009-NOV-5 by ORCHARD. 27-Oct-2007: Contacted by JYOTI. Plant J. (0960-7412) 2009 orchard Only protein-protein interactions imex curation Modulation of drought resistance by the abscisic acid-receptor PYL5 through inhibition of clade A PP2Cs. Modulation of drought resistance by the abscisic acid-receptor PYL5 through inhibition of clade A PP2Cs. Plant J. (0960-7412) 2009 imex curation Santiago J., Rodrigues A., Saez A., Rubio S., Antoni R., Dupeux F., Park SY., Marquez JA., Cutler SR., Rodriguez PL. prodiguez@ibmcp.upv.es 27-Oct-2007: 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 Santiago J., Rodrigues A., Saez A., Rubio S., Antoni R., Dupeux F., Park SY., Marquez JA., Cutler SR., Rodriguez PL. prodiguez@ibmcp.upv.es Accepted 2009-NOV-5 by ORCHARD. Plant J. (0960-7412) 2009 orchard Only protein-protein interactions imex curation Modulation of drought resistance by the abscisic acid-receptor PYL5 through inhibition of clade A PP2Cs. Modulation of drought resistance by the abscisic acid-receptor PYL5 through inhibition of clade A PP2Cs. Plant J. (0960-7412) 2009 imex curation Santiago J., Rodrigues A., Saez A., Rubio S., Antoni R., Dupeux F., Park SY., Marquez JA., Cutler SR., Rodriguez PL. prodiguez@ibmcp.upv.es 27-Oct-2007: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Santiago J., Rodrigues A., Saez A., Rubio S., Antoni R., Dupeux F., Park SY., Marquez JA., Cutler SR., Rodriguez PL. Note I could not curate the phosphatase assay as the Promega kit used contains two peptides and it is not clear if either or both are phosphorylated and how to represent this clearly to the user. Note I have omitted to include the binding of protein to non-natural enaiomer, as this might confuse users. We represent this data very crudely in IntAct at present and cannot clearly present the findings in an easily understandable manner. prodiguez@ibmcp.upv.es Accepted 2009-NOV-5 by ORCHARD. 27-Oct-2007: Contacted by JYOTI. Plant J. (0960-7412) 2009 orchard Only protein-protein interactions imex curation Modulation of drought resistance by the abscisic acid-receptor PYL5 through inhibition of clade A PP2Cs. Modulation of drought resistance by the abscisic acid-receptor PYL5 through inhibition of clade A PP2Cs. Plant J. (0960-7412) 2009 imex curation Santiago J., Rodrigues A., Saez A., Rubio S., Antoni R., Dupeux F., Park SY., Marquez JA., Cutler SR., Rodriguez PL. prodiguez@ibmcp.upv.es 27-Oct-2007: Contacted by JYOTI. orchard Only protein-protein interactions nicbe Nicotiana benthamiana bifc bimolecular fluorescence complementation bifc tag fluorescence tag visualisation by fluorescence tag fluorescence Santiago J., Rodrigues A., Saez A., Rubio S., Antoni R., Dupeux F., Park SY., Marquez JA., Cutler SR., Rodriguez PL. prodiguez@ibmcp.upv.es Accepted 2009-NOV-5 by ORCHARD. 27-Oct-2007: Contacted by JYOTI. Plant J. (0960-7412) 2009 orchard Only protein-protein interactions imex curation pyl5_arath Abscisic acid receptor PYL5 At5g05440 PYL5 ABIP3 RCAR8 PYR1-like protein 5 ABI1-binding protein 3 Regulatory components of ABA receptor 8 K18I23.25 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MRSPVQLQHGSDATNGFHTLQPHDQTDGPIKRVCLTRGMHVPEHVAMHHTHDVGPDQCCSSVVQMIHAPPESVWALVRRFDNPKVYKNFIRQCRIVQGDGLHVGDLREVMVVSGLPAVSSTERLEILDEERHVISFSVVGGDHRLKNYRSVTTLHASDDEGTVVVESYIVDVPPGNTEEETLSFVDTIVRCNLQSLARSTNRQ DIP protein Q9FLB1 original sequence version: 46 DIP protein Q9FLB1 original sequence version: 51 5A446F5E40D5AF11 pyr1_arath Abscisic acid receptor PYR1 PYR1 ABIP6 At4g17870 RCAR11 Protein PYRABACTIN RESISTANCE 1 ABI1-binding protein 6 Regulatory components of ABA receptor 11 T6K21.50 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MPSELTPEERSELKNSIAEFHTYQLDPGSCSSLHAQRIHAPPELVWSIVRRFDKPQTYKHFIKSCSVEQNFEMRVGCTRDVIVISGLPANTSTERLDILDDERRVTGFSIIGGEHRLTNYKSVTTVHRFEKENRIWTVVLESYVVDMPEGNSEDDTRMFADTVVKLNLQKLATVAEAMARNSGDGSGSQVT DIP protein O49686 original sequence version: 50 DIP protein O49686 original sequence version: 63 DIP protein O49686 original sequence version: 54 DIP protein O49686 original sequence version: 92 AD35DA240E286B55 abscisic acid abscisic acid small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) abscisic acid (CHEBI:22152) is a apo carotenoid sesquiterpenoids (CHEBI:36758) , abscisic acid (CHEBI:22152) is a monocarboxylic acids (CHEBI:25384) IUPAC name (2Z,4E)-5-(1-hydroxy-2,6,6-trimethyl-4-oxocyclohex-2-en-1-yl)-3-methylpenta-2,4-dienoic acid abscisic acid (CHEBI:22152) is a apo carotenoid sesquiterpenoids (CHEBI:36758) , abscisic acid (CHEBI:22152) is a monocarboxylic acids (CHEBI:25384) IUPAC name (2Z,4E)-5-(1-hydroxy-2,6,6-trimethyl-4-oxocyclohex-2-en-1-yl)-3-methylpenta-2,4-dienoic acid pyl8_arath Abscisic acid receptor PYL8 PYL8 ABIP1 RCAR3 PYR1-like protein 8 ABI1-binding protein 1 Regulatory components of ABA receptor 3 MFH8.10 At5g53160 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEANGIENLTNPNQEREFIRRHHKHELVDNQCSSTLVKHINAPVHIVWSLVRRFDQPQKYKPFISRCVVKGNMEIGTVREVDVKSGLPATRSTERLELLDDNEHILSIRIVGGDHRLKNYSSIISLHPETIEGRIGTLVIESFVVDVPEGNTKDETCYFVEALIKCNLKSLADISERLAVQDTTESRV D3436ADAC890AEED pyl4_arath Abscisic acid receptor PYL4 At2g38310 PYL4 ABIP2 RCAR10 PYR1-like protein 4 ABI1-binding protein 2 Regulatory components of ABA receptor 10 T19C21.20 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MLAVHRPSSAVSDGDSVQIPMMIASFQKRFPSLSRDSTAARFHTHEVGPNQCCSAVIQEISAPISTVWSVVRRFDNPQAYKHFLKSCSVIGGDGDNVGSLRQVHVVSGLPAASSTERLDILDDERHVISFSVVGGDHRLSNYRSVTTLHPSPISGTVVVESYVVDVPPGNTKEETCDFVDVIVRCNLQSLAKIAENTAAESKKKMSL DIP protein O80920 original sequence version: 50 DIP protein O80920 original sequence version: 54 DIP protein O80920 original sequence version: 68 118160B35D7066BB p2c16_arath Protein phosphatase 2C 16 HAB1 P2C-HA Protein phosphatase 2C HAB1 AtP2C-HA Protein HYPERSENSITIVE TO ABA 1 F28P22.4 At1g72770 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEEMTPAVAMTLSLAANTMCESSPVEITQLKNVTDAADLLSDSENQSFCNGGTECTMEDVSELEEVGEQDLLKTLSDTRSGSSNVFDEDDVLSVVEDNSAVISEGLLVVDAGSELSLSNTAMEIDNGRVLATAIIVGESSIEQVPTAEVLIAGVNQDTNTSEVVIRLPDENSNHLVKGRSVYELDCIPLWGTVSIQGNRSEMEDAFAVSPHFLKLPIKMLMGDHEGMSPSLTHLTGHFFGVYDGHGGHKVADYCRDRLHFALAEEIERIKDELCKRNTGEGRQVQWDKVFTSCFLTVDGEIEGKIGRAVVGSSDKVLEAVASETVGSTAVVALVCSSHIVVSNCGDSRAVLFRGKEAMPLSVDHKPDREDEYARIENAGGKVIQWQGARVFGVLAMSRSIGDRYLKPYVIPEPEVTFMPRSREDECLILASDGLWDVMNNQEVCEIARRRILMWHKKNGAPPLAERGKGIDPACQAAADYLSMLALQKGSKDNISIIVIDLKAQRKFKTRT DIP protein Q9CAJ0 original sequence version: 64 DIP protein Q9CAJ0-1 original sequence version: 62 DIP protein Q9CAJ0-1 original sequence version: 77 DIP protein Q9CAJ0 original sequence version: 62 DIP protein Q9CAJ0 original sequence version: 65 DIP protein Q9CAJ0-1 original sequence version: 66 DIP protein Q9CAJ0 original sequence version: 81 DIP protein Q9CAJ0-1 original sequence version: 86 DIP protein Q9CAJ0-1 original sequence version: 108 DIP protein Q9CAJ0 original sequence version: 112 DIP protein Q9CAJ0-1 original sequence version: 81 B001BAF0DD8333E1 pyl6_arath Abscisic acid receptor PYL6 T3G21 At2g40330 PYL6 ABIP5 RCAR9 PYR1-like protein 6 ABI1-binding protein 5 Regulatory components of ABA receptor 9 T7M7.15 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MPTSIQFQRSSTAAEAANATVRNYPHHHQKQVQKVSLTRGMADVPEHVELSHTHVVGPSQCFSVVVQDVEAPVSTVWSILSRFEHPQAYKHFVKSCHVVIGDGREVGSVREVRVVSGLPAAFSLERLEIMDDDRHVISFSVVGGDHRLMNYKSVTTVHESEEDSDGKKRTRVVESYVVDVPAGNDKEETCSFADTIVRCNLQSLAKLAENTSKFS DIP protein Q8S8E3 original sequence version: 35 DIP protein Q8S8E3 original sequence version: 40 8B9D3F7BD9E2860D hab1-pyl8 2 10 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 Q9CAJ0:p.Gly246Asp mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain G D Wed May 02 20:32:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly246asp' Tue Jun 30 19:28:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly246Asp' 8 unspecified role unspecified role prey prey physical association physical association Figure 1 hab1-pyl6 2 11 unspecified role unspecified role prey prey 10 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 Q9CAJ0:p.Gly246Asp mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain G D Wed May 02 15:37:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly246asp' Tue Jun 30 16:19:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly246Asp' physical association physical association Figure 1 hab1-pyl5 2 10 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 Q9CAJ0:p.Gly246Asp mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain G D Wed May 02 15:26:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly246asp' Tue Jun 30 18:51:56 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly246Asp' 5 unspecified role unspecified role prey prey physical association physical association Figure 1 hab1-pyl6-1 2 10 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 11 unspecified role unspecified role bait bait physical association physical association Figure 1 hab1-pyl8-1 2 8 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey Q9CAJ0:p.Gly246Asp mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain G D Wed May 02 14:59:21 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly246asp' Tue Jun 30 19:34:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly246Asp' physical association physical association Figure 1 hab1-pyl5-1 2 10 unspecified role unspecified role prey prey Q9CAJ0:p.Gly246Asp mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain G D Wed May 02 17:45:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly246asp' Tue Jun 30 16:29:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly246Asp' 5 unspecified role unspecified role bait bait physical association physical association Figure 1 hab1-pyl4 2 10 unspecified role unspecified role prey prey Q9CAJ0:p.Gly246Asp mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain G D Wed May 02 15:31:26 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly246asp' Tue Jun 30 15:36:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly246Asp' 9 unspecified role unspecified role bait bait physical association physical association Figure 1 The interaction only occurred in the presence of 10 micro molar abscisic acid hab1-pyr1 2 6 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey Q9CAJ0:p.Gly246Asp mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain G D Wed May 02 14:49:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly246asp' Tue Jun 30 20:19:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly246Asp' physical association physical association Figure 1 The interaction only occurred in the presence of 10 micro molar abscisic acid hab1-pyl5-2 4 10 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 Q9CAJ0:p.Gly246Asp mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain G D Wed May 02 19:32:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly246asp' Tue Jun 30 16:36:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly246Asp' region N-terminal region of 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 5 unspecified role unspecified role neutral component neutral component region C-terminal region of 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 physical association physical association Figure 3d Bimolecular fluorescence complementation (BiFC) assays were used to test the interaction between HAB1 and PYL5 in plant cells. hab1-pyl5-3 1 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 region N-terminal region of 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 5 unspecified role unspecified role prey prey region C-terminal region of 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 physical association physical association Figure 3e We found that HAB1 and PYL5 co-immunoprecipitated, as YFPC-PYL5 was detected in the immunocomplex precipitated with an antibody to the myc epitope, which pulls down the HAB1-myc-YFPN fusion protein (Figure 3e). pyl5-aba 3 7 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 7 Dissociation constant 1.1 micro molar , enthalpy value -7.9Kcal -1 mol, entropy value 1.3 cal mol-1 K-1 addition of protein Q9CAJ0 C-terminal domain (amino acid region 179 to 511).