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/ Detection of protein-protein interactions in plants using bimolecular fluorescence complementation. Detection of protein-protein interactions in plants using bimolecular fluorescence complementation. Plant J. (0960-7412) 2004 imex curation Bracha-Drori K., Shichrur K., Katz A., Oliva M., Angelovici R., Yalovsky S., Ohad N. To allow expression of YN and YC fusion proteins in plants, corresponding genes were cloned between the CaMV 35S promoter and NOS transcriptional terminator. The entire YN and YC expression cassettes were in turn subcloned into T-DNA base plant binary vectors. Reconstitution of YFP fluorescence was examined by transient co-expression of the tested protein pairs. Plants were transformed by injecting Agrobacterium tumefaciens cells harboring the appropriate plasmid to the abaxial side of Nicotiana benthamiana leaves. shauly@tauex.tau.ac.il 16-April-2008: Contacted by JYOTI. Only protein-protein interactions tobac Nicotiana tabacum (Common tobacco) Common tobacco bifc bimolecular fluorescence complementation bifc predetermined predetermined participant predetermined Bracha-Drori K., Shichrur K., Katz A., Oliva M., Angelovici R., Yalovsky S., Ohad N. To allow expression of YN and YC fusion proteins in plants, corresponding genes were cloned between the CaMV 35S promoter and NOS transcriptional terminator. The entire YN and YC expression cassettes were in turn subcloned into T-DNA base plant binary vectors. Reconstitution of YFP fluorescence was examined by transient co-expression of the tested protein pairs. Plants were transformed by injecting Agrobacterium tumefaciens cells harboring the appropriate plasmid to the abaxial side of Nicotiana benthamiana leaves. shauly@tauex.tau.ac.il Accepted 2008-APR-7 by CDEROW. Plant J. (0960-7412) 2004 Only protein-protein interactions imex curation Detection of protein-protein interactions in plants using bimolecular fluorescence complementation. Detection of protein-protein interactions in plants using bimolecular fluorescence complementation. Plant J. (0960-7412) 2004 imex curation Bracha-Drori K., Shichrur K., Katz A., Oliva M., Angelovici R., Yalovsky S., Ohad N. To allow expression of YN and YC fusion proteins in plants, corresponding genes were cloned between the CaMV 35S promoter and NOS transcriptional terminator. The entire YN and YC expression cassettes were in turn subcloned into T-DNA base plant binary vectors. Reconstitution of YFP fluorescence was examined by transient co-expression of the tested protein pairs. Plants were transformed by injecting Agrobacterium tumefaciens cells harboring the appropriate plasmid to the abaxial side of Nicotiana benthamiana leaves. shauly@tauex.tau.ac.il 16-April-2008: Contacted by JYOTI. Only protein-protein interactions arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Bracha-Drori K., Shichrur K., Katz A., Oliva M., Angelovici R., Yalovsky S., Ohad N. shauly@tauex.tau.ac.il Accepted 2008-APR-7 by CDEROW. 16-April-2008: Contacted by JYOTI. Plant J. (0960-7412) 2004 Only protein-protein interactions imex curation fnta_arath Protein farnesyltransferase/geranylgeranyltransferase type-1 subunit alpha CAAX farnesyltransferase subunit alpha Ras proteins prenyltransferase subunit alpha FTase-alpha Type I protein geranyl-geranyltransferase subunit alpha F25L23.240 At3g59380 FTA protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MNFDETVPLSQRLEWSDVVPLTQDDGPNPVVPIAYKEEFRETMDYFRAIYFSDERSPRALRLTEETLLLNSGNYTVWHFRRLVLEALNHDLFEELEFIERIAEDNSKNYQLWHHRRWVAEKLGPDVAGRELEFTRRVLSLDAKHYHAWSHRQWTLRALGGWEDELDYCHELLEADVFNNSAWNQRYYVITQSPLLGGLEAMRESEVSYTIKAILTNPANESSWRYLKALYKDDKESWISDPSVSSVCLNVLSRTDCFHGFALSTLLDLLCDGLRPTNEHKDSVRALANEEPETNLANLVCTILGRVDPVRANYWAWRKSKITVAAI 78146021EAFC785A fie_arath Polycomb group protein FERTILIZATION-INDEPENDENT ENDOSPERM FIE FIS3 At3g20740 MOE17.5 Protein FERTILIZATION-INDEPENDENT SEED 3 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSKITLGNESIVGSLTPSNKKSYKVTNRIQEGKKPLYAVVFNFLDARFFDVFVTAGGNRITLYNCLGDGAISALQSYADEDKEESFYTVSWACGVNGNPYVAAGGVKGIIRVIDVNSETIHKSLVGHGDSVNEIRTQPLKPQLVITASKDESVRLWNVETGICILIFAGAGGHRYEVLSVDFHPSDIYRFASCGMDTTIKIWSMKEFWTYVEKSFTWTDDPSKFPTKFVQFPVFTASIHTNYVDCNRWFGDFILSKSVDNEILLWEPQLKENSPGEGASDVLLRYPVPMCDIWFIKFSCDLHLSSVAIGNQEGKVYVWDLKSCPPVLITKLSHNQSKSVIRQTAMSVDGSTILACCEDGTIWRWDVITK DIP protein Q9LT47 original sequence version: 126 DIP protein Q9LT47 original sequence version: 91 EDF0989C02B4FD4A medea_arath Histone-lysine N-methyltransferase MEDEA EMB173 Maternal embryogenesis control protein Protein FERTILIZATION-INDEPENDENT SEED 1 Protein SET DOMAIN GROUP 5 Protein EMBRYO DEFECTIVE 173 MEA FIS1 At1g02580 T14P4.11 MEDEA SDG5 SET5 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEKENHEDDGEGLPPELNQIKEQIEKERFLHIKRKFELRYIPSVATHASHHQSFDLNQPAAEDDNGGDNKSLLSRMQNPLRHFSASSDYNSYEDQGYVLDEDQDYALEEDVPLFLDEDVPLLPSVKLPIVEKLPRSITWVFTKSSQLMAESDSVIGKRQIYYLNGEALELSSEEDEEDEEEDEEEIKKEKCEFSEDVDRFIWTVGQDYGLDDLVVRRALAKYLEVDVSDILERYNELKLKNDGTAGEASDLTSKTITTAFQDFADRRHCRRCMIFDCHMHEKYEPESRSSEDKSSLFEDEDRQPCSEHCYLKVRSVTEADHVMDNDNSISNKIVVSDPNNTMWTPVEKDLYLKGIEIFGRNSCDVALNILRGLKTCLEIYNYMREQDQCTMSLDLNKTTQRHNQVTKKVSRKSSRSVRKKSRLRKYARYPPALKKTTSGEAKFYKHYTPCTCKSKCGQQCPCLTHENCCEKYCGCSKDCNNRFGGCNCAIGQCTNRQCPCFAANRECDPDLCRSCPLSCGDGTLGETPVQIQCKNMQFLLQTNKKILIGKSDVHGWGAFTWDSLKKNEYLGEYTGELITHDEANERGRIEDRIGSSYLFTLNDQLEIDARRKGNEFKFLNHSARPNCYAKLMIVRGDQRIGLFAERAIEEGEELFFDYCYGPEHADWSRGREPRKTGASKRSKEARPAR 2103D06ACD589CDF fntb_arath Protein farnesyltransferase subunit beta CAAX farnesyltransferase subunit beta Ras proteins prenyltransferase subunit beta Enhanced response to abscisic acid 1 FTB ERA1 WIGGUM MSN9.20 At5g40280 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MPVVTRLIRLKCVGLRLDRSGLNRRICHGGHGESTRRRVMEELSSLTVSQREQFLVENDVFGIYNYFDASDVSTQKYMMEIQRDKQLDYLMKGLRQLGPQFSSLDANRPWLCYWILHSIALLGETVDDELESNAIDFLGRCQGSEGGYGGGPGQLPHLATTYAAVNALVTLGGDKALSSINREKMSCFLRRMKDTSGGFRMHDMGEMDVRACYTAISVASILNIMDDELTQGLGDYILSCQTYEGGIGGEPGSEAHGGYTYCGLAAMILINEVDRLNLDSLMNWAVHRQGVEMGFQGRTNKLVDGCYTFWQAAPCVLLQRLYSTNDHDVHGSSHISEGTNEEHHAHDEDDLEDSDDDDDSDEDNDEDSVNGHRIHHTSTYINRRMQLVFDSLGLQRYVLLCSKIPDGGFRDKPRKPRDFYHTCYCLSGLSVAQHAWLKDEDTPPLTRDIMGGYSNLLEPVQLLHNIVMDQYNEAIEFFFKAA 4A557EB53567C83B pfta-pftb 1 6 unspecified role unspecified role neutral component neutral component n-terminal C terminal region of the YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 3 unspecified role unspecified role neutral component neutral component n-terminal N terminal region of YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range direct interaction direct interaction 2a Interaction between FTA and FTB fused to either YN or YC resulted in similar fluorescence intensity as long as they were fused downstream of the YFP domain (Figure 2a,b). YFP fluorescence was substantially weaker when FTA and FTB were fused to YN and YC at opposite orientations, upstream and downstream of the split YFP domains (Figure 2c). No fluorescence was detected when FTA and FTB were fused at the same orientation, upstream of YN and YC pfta-pftb-1 1 6 unspecified role unspecified role neutral component neutral component n-terminal N terminal region of the YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 3 unspecified role unspecified role neutral component neutral component n-terminal C terminal region of YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range direct interaction direct interaction 2b Interaction between FTA and FTB fused to either YN or YC resulted in similar fluorescence intensity as long as they were fused downstream of the YFP domain (Figure 2a,b). YFP fluorescence was substantially weaker when FTA and FTB were fused to YN and YC at opposite orientations, upstream and downstream of the split YFP domains (Figure 2c). No fluorescence was detected when FTA and FTB were fused at the same orientation, upstream of YN and YC pfta-pftb-2 1 6 unspecified role unspecified role neutral component neutral component c-terminal N-terminal region of the YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 3 unspecified role unspecified role neutral component neutral component n-terminal C-terminal region of the YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range direct interaction direct interaction 2c Interaction between FTA and FTB fused to either YN or YC resulted in similar fluorescence intensity as long as they were fused downstream of the YFP domain (Figure 2a,b). YFP fluorescence was substantially weaker when FTA and FTB were fused to YN and YC at opposite orientations, upstream and downstream of the split YFP domains (Figure 2c). No fluorescence was detected when FTA and FTB were fused at the same orientation, upstream of YN and YC mea-fie-2 1 5 unspecified role unspecified role neutral component neutral component n-terminal N terminal region of YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 4 unspecified role unspecified role neutral component neutral component n-terminal C terminal region of YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association 2d The fluorescence intensity obtained from the interaction between FIE and MEA pfta-pftb-3 2 6 unspecified role unspecified role prey prey n-terminal N terminal region of the YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 3 unspecified role unspecified role bait bait n-terminal C terminal region of YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association 4b Co-immunoprecipitation experiments (Figure 4b) further demonstrated the interaction between FTA and FTB as well as FIE and MEA in the BiFC. mea-fie-3 2 4 unspecified role unspecified role bait bait n-terminal C terminal region of YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role prey prey n-terminal N terminal region of YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association 4b Co-immunoprecipitation experiments (Figure 4b) further demonstrated the interaction between FTA and FTB as well as FIE and MEA in the BiFC.