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 CRY1 blue light photoreceptor of Arabidopsis interacts with phytochrome A in vitro. The CRY1 blue light photoreceptor of Arabidopsis interacts with phytochrome A in vitro. Mol. Cell (1097-2765) 1998 imex curation Ahmad M., Jarillo JA., Smirnova O., Cashmore AR. mahmad@sas.upenn.edu 21-Dec-2010: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Ahmad M., Jarillo JA., Smirnova O., Cashmore AR. mahmad@sas.upenn.edu Accepted 2009-JUN-5 by ORCHARD. 21-Dec-2010: Contacted by JYOTI. Mol. Cell (1097-2765) 1998 orchard Only protein-protein interactions imex curation The CRY1 blue light photoreceptor of Arabidopsis interacts with phytochrome A in vitro. The CRY1 blue light photoreceptor of Arabidopsis interacts with phytochrome A in vitro. Mol. Cell (1097-2765) 1998 imex curation Ahmad M., Jarillo JA., Smirnova O., Cashmore AR. mahmad@sas.upenn.edu 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 Ahmad M., Jarillo JA., Smirnova O., Cashmore AR. mahmad@sas.upenn.edu Accepted 2009-JUN-5 by ORCHARD. Mol. Cell (1097-2765) 1998 orchard Only protein-protein interactions imex curation phya_arath Phytochrome A PHYA HY8 At1g09570 F14J9.23 FHY2 FRE1 Protein FAR RED ELONGATED HYPOCOTYL 2 Protein FAR RED ELONGATED 1 Protein ELONGATED HYPOCOTYL 8 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSGSRPTQSSEGSRRSRHSARIIAQTTVDAKLHADFEESGSSFDYSTSVRVTGPVVENQPPRSDKVTTTYLHHIQKGKLIQPFGCLLALDEKTFKVIAYSENASELLTMASHAVPSVGEHPVLGIGTDIRSLFTAPSASALQKALGFGDVSLLNPILVHCRTSAKPFYAIIHRVTGSIIIDFEPVKPYEVPMTAAGALQSYKLAAKAITRLQSLPSGSMERLCDTMVQEVFELTGYDRVMAYKFHEDDHGEVVSEVTKPGLEPYLGLHYPATDIPQAARFLFMKNKVRMIVDCNAKHARVLQDEKLSFDLTLCGSTLRAPHSCHLQYMANMDSIASLVMAVVVNEEDGEGDAPDATTQPQKRKRLWGLVVCHNTTPRFVPFPLRYACEFLAQVFAIHVNKEVELDNQMVEKNILRTQTLLCDMLMRDAPLGIVSQSPNIMDLVKCDGAALLYKDKIWKLGTTPSEFHLQEIASWLCEYHMDSTGLSTDSLHDAGFPRALSLGDSVCGMAAVRISSKDMIFWFRSHTAGEVRWGGAKHDPDDRDDARRMHPRSSFKAFLEVVKTRSLPWKDYEMDAIHSLQLILRNAFKDSETTDVNTKVIYSKLNDLKIDGIQELEAVTSEMVRLIETATVPILAVDSDGLVNGWNTKIAELTGLSVDEAIGKHFLTLVEDSSVEIVKRMLENALEGTEEQNVQFEIKTHLSRADAGPISLVVNACASRDLHENVVGVCFVAHDLTGQKTVMDKFTRIEGDYKAIIQNPNPLIPPIFGTDEFGWCTEWNPAMSKLTGLKREEVIDKMLLGEVFGTQKSCCRLKNQEAFVNLGIVLNNAVTSQDPEKVSFAFFTRGGKYVECLLCVSKKLDREGVVTGVFCFLQLASHELQQALHVQRLAERTAVKRLKALAYIKRQIRNPLSGIMFTRKMIEGTELGPEQRRILQTSALCQKQLSKILDDSDLESIIEGCLDLEMKEFTLNEVLTASTSQVMMKSNGKSVRITNETGEEVMSDTLYGDSIRLQQVLADFMLMAVNFTPSGGQLTVSASLRKDQLGRSVHLANLEIRLTHTGAGIPEFLLNQMFGTEEDVSEEGLSLMVSRKLVKLMNGDVQYLRQAGKSSFIITAELAAANK DIP protein P14712 original sequence version: 133 D8B359713430744B phya3_avesa Phytochrome A type 3 PHYA3 PHY3 protein protein avesa Avena sativa (Oat) MSSSRPASSSSSRNRQSSQARVLAQTTLDAELNAEYEESGDSFDYSKLVEAQRDGPPVQQGRSEKVIAYLQHIQKGKLIQTFGCLLALDEKSFNVIAFSENAPEMLTTVSHAVPSVDDPPRLGIGTNVRSLFSDQGATALHKALGFADVSLLNPILVQCKTSGKPFYAIVHRATGCLVVDFEPVKPTEFPATAAGALQSYKLAAKAISKIQSLPGGSMEVLCNTVVKEVFDLTGYDRVMAYKFHEDDHGEVFSEITKPGLEPYLGLHYPATDIPQAARLLFMKNKVRMICDCRARSIKVIEAEALPFDISLCGSALRAPHSCHLQYMENMNSIASLVMAVVVNENEEDDEAESEQPAQQQKKKKLWGLLVCHHESPRYVPFPLRYACEFLAQVFAVHVNREFELEKQLREKNILKMQTMLSDMLFREASPLTIVSGTPNIMDLVKCDGAALLYGGKVWRLRNAPTESQIHDIAFWLSDVHRDSTGLSTDSLHDAGYPGAAALGDMICGMAVAKINSKDILFWFRSHTAAEIRWGGAKNDPSDMDDSRRMHPRLSFKAFLEVVKMKSLPWSDYEMDAIHSLQLILRGTLNDASKPKREASLDNQIGDLKLDGLAELQAVTSEMVRLMETATVPILAVDGNGLVNGWNQKAAELTGLRVDDAIGRHILTLVEDSSVPVVQRMLYLALQGKEEKEVRFEVKTHGPKRDDGPVILVVNACASRDLHDHVVGVCFVAQDMTVHKLVMDKFTRVEGDYKAIIHNPNPLIPPIFGADEFGWCSEWNAAMTKLTGWNRDEVLDKMLLGEVFDSSNASCPLKNRDAFVSLCVLINSALAGEETEKAPFGFFDRSGKYIECLLSANRKENEGGLITGVFCFIHVASHELQHALQVQQASEQTSLKRLKAFSYMRHAINNPLSGMLYSRKALKNTDLNEEQMKQIHVGDNCHHQINKILADLDQDSITEKSSCLDLEMAEFLLQDVVVAAVSQVLITCQGKGIRISCNLPERFMKQSVYGDGVRLQQILSDFLFISVKFSPVGGSVEISSKLTKNSIGENLHLIDLELRIKHQGLGVPAELMAQMFEEDNKEQSEEGLSLLVSRNLLRLMNGDVRHLREAGVSTFIITAELASAPTAMGQ 8AC1083533567ACF cry1_arath Cryptochrome-1 CRY1 HY4 At4g08920 T3H13.5 T3H13.14 BLU1 Blue light photoreceptor Protein OUT OF PHASE 2 Protein BLUE LIGHT UNINHIBITED 1 Protein ELONGATED HYPOCOTYL 4 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSGSVSGCGSGGCSIVWFRRDLRVEDNPALAAAVRAGPVIALFVWAPEEEGHYHPGRVSRWWLKNSLAQLDSSLRSLGTCLITKRSTDSVASLLDVVKSTGASQIFFNHLYDPLSLVRDHRAKDVLTAQGIAVRSFNADLLYEPWEVTDELGRPFSMFAAFWERCLSMPYDPESPLLPPKKIISGDVSKCVADPLVFEDDSEKGSNALLARAWSPGWSNGDKALTTFINGPLLEYSKNRRKADSATTSFLSPHLHFGEVSVRKVFHLVRIKQVAWANEGNEAGEESVNLFLKSIGLREYSRYISFNHPYSHERPLLGHLKFFPWAVDENYFKAWRQGRTGYPLVDAGMRELWATGWLHDRIRVVVSSFFVKVLQLPWRWGMKYFWDTLLDADLESDALGWQYITGTLPDSREFDRIDNPQFEGYKFDPNGEYVRRWLPELSRLPTDWIHHPWNAPESVLQAAGIELGSNYPLPIVGLDEAKARLHEALSQMWQLEAASRAAIENGSEEGLGDSAEVEEAPIEFPRDITMEETEPTRLNPNRRYEDQMVPSITSSLIRPEEDEESSLNLRNSVGDSRAEVPRNMVNTNQAQQRRAEPASNQVTAMIPEFNIRIVAESTEDSTAESSSSGRRERSGGIVPEWSPGYSEQFPSEENGIGGGSTTSSYLQNHHEILNWRRLSQTG 372A7E6DDC2AC076 phya-cry1-1 2 5 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 3 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 10 8 physical association physical association Figure 3 However, when the larger PHYA-containing constructs (BD Phy 624-1100) were cotransformed with any of the pACT constructs containing cryptochrome sequences, a significant level of β-galactosidase activity was detectable (Figure 3B). The induction was more than 20-fold over background levels; this level of activity is indicative of a direct interaction between CRY1 and phytochrome protein fragments in living yeast cells. phya-cry1-2 1 5 enzyme target enzyme target substrate substrate neutral component neutral component spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm 4 enzyme enzyme neutral component neutral component yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast phosphorylation phosphorylation reaction phosphorylation Figure 1 Phosphorylation of CRY1 by Phytochrome-Associated Kinase Activity In Vitro phosphorylation of Cry1 pyrophosphate phya-cry1-3 1 4 enzyme enzyme neutral component neutral component yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 5 enzyme target enzyme target substrate substrate neutral component neutral component c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Figure 2 Phosphorylation of mutant CRY1 C-terminal fragments by phytochrome in vitro phosphorylation of Cry1 details of purification of CRY1 from E.coli are not provided by in the paper. The authors do not provide the amino acid coordinates of the mutants used in the study , therefore where these mutants effected phosphorylation this data could not be curated. phya-3 1 4 self self neutral component neutral component yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast phosphorylation phosphorylation reaction phosphorylation true Figure 1 Addition of phytochrome alone to the phosphorylation reactions resulted in the appearance of a phosphorylated species (approximately 120 kDa) that comigrated with phytochrome; this species represents autophosphorylated phytochrome, autophosphoryllation