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/ Obligate Heterodimerization of Arabidopsis Phytochromes C and E and Interaction with the PIF3 Basic Helix-Loop-Helix Transcription Factor. Obligate Heterodimerization of Arabidopsis Phytochromes C and E and Interaction with the PIF3 Basic Helix-Loop-Helix Transcription Factor. Plant Cell (1040-4651) 2009 imex curation Clack T., Shokry A., Moffet M., Liu P., Faul M., Sharrock RA. sharrock@montana.edu 10-Jul-2009: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro comigration in gel comigration in gel electrophoresis comigration in gel autoradiography autoradiography Clack T., Shokry A., Moffet M., Liu P., Faul M., Sharrock RA. sharrock@montana.edu Accepted 2009-JUN-8 by ORCHARD. 10-Jul-2009: Contacted by JYOTI. Plant Cell (1040-4651) 2009 orchard Only protein-protein interactions imex curation Obligate Heterodimerization of Arabidopsis Phytochromes C and E and Interaction with the PIF3 Basic Helix-Loop-Helix Transcription Factor. Obligate Heterodimerization of Arabidopsis Phytochromes C and E and Interaction with the PIF3 Basic Helix-Loop-Helix Transcription Factor. Plant Cell (1040-4651) 2009 imex curation Clack T., Shokry A., Moffet M., Liu P., Faul M., Sharrock RA. sharrock@montana.edu 10-Jul-2009: 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 Clack T., Shokry A., Moffet M., Liu P., Faul M., Sharrock RA. sharrock@montana.edu Accepted 2009-JUN-8 by ORCHARD. Plant Cell (1040-4651) 2009 orchard Only protein-protein interactions imex curation Obligate Heterodimerization of Arabidopsis Phytochromes C and E and Interaction with the PIF3 Basic Helix-Loop-Helix Transcription Factor. Obligate Heterodimerization of Arabidopsis Phytochromes C and E and Interaction with the PIF3 Basic Helix-Loop-Helix Transcription Factor. Plant Cell (1040-4651) 2009 imex curation Clack T., Shokry A., Moffet M., Liu P., Faul M., Sharrock RA. sharrock@montana.edu 10-Jul-2009: Contacted by JYOTI. orchard Only protein-protein interactions arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Clack T., Shokry A., Moffet M., Liu P., Faul M., Sharrock RA. Transgenic Arabidopsis seedligs expressing MYC tagged or HIS tagged PhyC protein, or MYC tagged PhyB or PHYE proteins were used in this study. sharrock@montana.edu Accepted 2009-JUN-8 by ORCHARD. 10-Jul-2009: Contacted by JYOTI. Plant Cell (1040-4651) 2009 orchard Only protein-protein interactions imex curation Obligate Heterodimerization of Arabidopsis Phytochromes C and E and Interaction with the PIF3 Basic Helix-Loop-Helix Transcription Factor. Obligate Heterodimerization of Arabidopsis Phytochromes C and E and Interaction with the PIF3 Basic Helix-Loop-Helix Transcription Factor. Plant Cell (1040-4651) 2009 imex curation Clack T., Shokry A., Moffet M., Liu P., Faul M., Sharrock RA. sharrock@montana.edu 10-Jul-2009: Contacted by JYOTI. orchard Only protein-protein interactions arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Clack T., Shokry A., Moffet M., Liu P., Faul M., Sharrock RA. Transgenic Arabidopsis seedligs expressing MYC tagged PIF3 protein were used in this study. The dark grown seedlings were treated with a pulse of red light (670nm) at 30 micro-moles per meter squared per second for 30 seconds prior to coip assay. sharrock@montana.edu Accepted 2009-JUN-8 by ORCHARD. 10-Jul-2009: Contacted by JYOTI. Plant Cell (1040-4651) 2009 orchard Only protein-protein interactions imex curation phyb_arath Phytochrome B PHYB HY3 OOP1 Protein LONG HYPOCOTYL 3 Protein OUT OF PHASE 1 MSF3.17 At2g18790 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MVSGVGGSGGGRGGGRGGEEEPSSSHTPNNRRGGEQAQSSGTKSLRPRSNTESMSKAIQQYTVDARLHAVFEQSGESGKSFDYSQSLKTTTYGSSVPEQQITAYLSRIQRGGYIQPFGCMIAVDESSFRIIGYSENAREMLGIMPQSVPTLEKPEILAMGTDVRSLFTSSSSILLERAFVAREITLLNPVWIHSKNTGKPFYAILHRIDVGVVIDLEPARTEDPALSIAGAVQSQKLAVRAISQLQALPGGDIKLLCDTVVESVRDLTGYDRVMVYKFHEDEHGEVVAESKRDDLEPYIGLHYPATDIPQASRFLFKQNRVRMIVDCNATPVLVVQDDRLTQSMCLVGSTLRAPHGCHSQYMANMGSIASLAMAVIINGNEDDGSNVASGRSSMRLWGLVVCHHTSSRCIPFPLRYACEFLMQAFGLQLNMELQLALQMSEKRVLRTQTLLCDMLLRDSPAGIVTQSPSIMDLVKCDGAAFLYHGKYYPLGVAPSEVQIKDVVEWLLANHADSTGLSTDSLGDAGYPGAAALGDAVCGMAVAYITKRDFLFWFRSHTAKEIKWGGAKHHPEDKDDGQRMHPRSSFQAFLEVVKSRSQPWETAEMDAIHSLQLILRDSFKESEAAMNSKVVDGVVQPCRDMAGEQGIDELGAVAREMVRLIETATVPIFAVDAGGCINGWNAKIAELTGLSVEEAMGKSLVSDLIYKENEATVNKLLSRALRGDEEKNVEVKLKTFSPELQGKAVFVVVNACSSKDYLNNIVGVCFVGQDVTSQKIVMDKFINIQGDYKAIVHSPNPLIPPIFAADENTCCLEWNMAMEKLTGWSRSEVIGKMIVGEVFGSCCMLKGPDALTKFMIVLHNAIGGQDTDKFPFPFFDRNGKFVQALLTANKRVSLEGKVIGAFCFLQIPSPELQQALAVQRRQDTECFTKAKELAYICQVIKNPLSGMRFANSLLEATDLNEDQKQLLETSVSCEKQISRIVGDMDLESIEDGSFVLKREEFFLGSVINAIVSQAMFLLRDRGLQLIRDIPEEIKSIEVFGDQIRIQQLLAEFLLSIIRYAPSQEWVEIHLSQLSKQMADGFAAIRTEFRMACPGEGLPPELVRDMFHSSRWTSPEGLGLSVCRKILKLMNGEVQYIRESERSYFLIILELPVPRKRPLSTASGSGDMMLMMPY DIP protein P14713 original sequence version: 107 DIP protein P14713 original sequence version: 131 7B5348CB1091B813 pif3_arath Transcription factor PIF3 Phytochrome-associated protein 3 Basic helix-loop-helix protein 8 PIF3 PAP3 BHLH8 At1g09530 F14J9.19 Transcription factor EN 100 bHLH transcription factor bHLH008 Phytochrome-interacting factor 3 EN100 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MPLFELFRLTKAKLESAQDRNPSPPVDEVVELVWENGQISTQSQSSRSRNIPPPQANSSRAREIGNGSKTTMVDEIPMSVPSLMTGLSQDDDFVPWLNHHPSLDGYCSDFLRDVSSPVTVNEQESDMAVNQTAFPLFQRRKDGNESAPAASSSQYNGFQSHSLYGSDRARDLPSQQTNPDRFTQTQEPLITSNKPSLVNFSHFLRPATFAKTTNNNLHDTKEKSPQSPPNVFQTRVLGAKDSEDKVLNESVASATPKDNQKACLISEDSCRKDQESEKAVVCSSVGSGNSLDGPSESPSLSLKRKHSNIQDIDCHSEDVEEESGDGRKEAGPSRTGLGSKRSRSAEVHNLSERRRRDRINEKMRALQELIPNCNKVDKASMLDEAIEYLKSLQLQVQIMSMASGYYLPPAVMFPPGMGHYPAAAAAMAMGMGMPYAMGLPDLSRGGSSVNHGPQFQVSGMQQQPVAMGIPRVSGGGIFAGSSTIGNGSTRDLSGSKDQTTTNNNSNLKPIKRKQGSSDQFCGSS DIP protein O80536 original sequence version: 116 DIP protein O80536 original sequence version: 85 1044AC01D598DE7C phye_arath Phytochrome E PHYE At4g18130 F15J5.100 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGFESSSSAASNMKPQPQKSNTAQYSVDAALFADFAQSIYTGKSFNYSKSVISPPNHVPDEHITAYLSNIQRGGLVQPFGCLIAVEEPSFRILGLSDNSSDFLGLLSLPSTSHSGEFDKVKGLIGIDARTLFTPSSGASLSKAASFTEISLLNPVLVHSRTTQKPFYAILHRIDAGIVMDLEPAKSGDPALTLAGAVQSQKLAVRAISRLQSLPGGDIGALCDTVVEDVQRLTGYDRVMVYQFHEDDHGEVVSEIRRSDLEPYLGLHYPATDIPQAARFLFKQNRVRMICDCNATPVKVVQSEELKRPLCLVNSTLRAPHGCHTQYMANMGSVASLALAIVVKGKDSSKLWGLVVGHHCSPRYVPFPLRYACEFLMQAFGLQLQMELQLASQLAEKKAMRTQTLLCDMLLRDTVSAIVTQSPGIMDLVKCDGAALYYKGKCWLVGVTPNESQVKDLVNWLVENHGDDSTGLTTDSLVDAGYPGAISLGDAVCGVAAAGFSSKDYLLWFRSNTASAIKWGGAKHHPKDKDDAGRMHPRSSFTAFLEVAKSRSLPWEISEIDAIHSLRLIMRESFTSSRPVLSGNGVARDANELTSFVCEMVRVIETATAPIFGVDSSGCINGWNKKTAEMTGLLASEAMGKSLADEIVQEESRAALESLLCKALQGEEEKSVMLKLRKFGQNNHPDYSSDVCVLVNSCTSRDYTENIIGVCFVGQDITSEKAITDRFIRLQGDYKTIVQSLNPLIPPIFASDENACCSEWNAAMEKLTGWSKHEVIGKMLPGEVFGVFCKVKCQDSLTKFLISLYQGIAGDNVPESSLVEFFNKEGKYIEASLTANKSTNIEGKVIRCFFFLQIINKESGLSCPELKESAQSLNELTYVRQEIKNPLNGIRFAHKLLESSEISASQRQFLETSDACEKQITTIIESTDLKSIEEGKLQLETEEFRLENILDTIISQVMIILRERNSQLRVEVAEEIKTLPLNGDRVKLQLILADLLRNIVNHAPFPNSWVGISISPGQELSRDNGRYIHLQFRMIHPGKGLPSEMLSDMFETRDGWVTPDGLGLKLSRKLLEQMNGRVSYVREDERCFFQVDLQVKTMLGVESRGTEGSSSIK 2E06677039213B57 phyd_arath Phytochrome D PHYD At4g16250 dl4165c protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MVSGGGSKTSGGEAASSGHRRSRHTSAAEQAQSSANKALRSQNQQPQNHGGGTESTNKAIQQYTVDARLHAVFEQSGESGKSFDYSQSLKTAPYDSSVPEQQITAYLSRIQRGGYTQPFGCLIAVEESTFTIIGYSENAREMLGLMSQSVPSIEDKSEVLTIGTDLRSLFKSSSYLLLERAFVAREITLLNPIWIHSNNTGKPFYAILHRVDVGILIDLEPARTEDPALSIAGAVQSQKLAVRAISHLQSLPSGDIKLLCDTVVESVRDLTGYDRVMVYKFHEDEHGEVVAESKRNDLEPYIGLHYPATDIPQASRFLFKQNRVRMIVDCYASPVRVVQDDRLTQFICLVGSTLRAPHGCHAQYMTNMGSIASLAMAVIINGNEEDGNGVNTGGRNSMRLWGLVVCHHTSARCIPFPLRYACEFLMQAFGLQLNMELQLALQVSEKRVLRMQTLLCDMLLRDSPAGIVTQRPSIMDLVKCNGAAFLYQGKYYPLGVTPTDSQINDIVEWLVANHSDSTGLSTDSLGDAGYPRAAALGDAVCGMAVACITKRDFLFWFRSHTEKEIKWGGAKHHPEDKDDGQRMNPRSSFQTFLEVVKSRCQPWETAEMDAIHSLQLILRDSFKESEAMDSKAAAAGAVQPHGDDMVQQGMQEIGAVAREMVRLIETATVPIFAVDIDGCINGWNAKIAELTGLSVEDAMGKSLVRELIYKEYKETVDRLLSCALKGDEGKNVEVKLKTFGSELQGKAMFVVVNACSSKDYLNNIVGVCFVGQDVTGHKIVMDKFINIQGDYKAIIHSPNPLIPPIFAADENTCCLEWNTAMEKLTGWPRSEVIGKLLVREVFGSYCRLKGPDALTKFMIVLHNAIGGQDTDKFPFPFFDRKGEFIQALLTLNKRVSIDGKIIGAFCFLQIPSPELQQALEVQRRQESEYFSRRKELAYIFQVIKNPLSGLRFTNSLLEDMDLNEDQKQLLETSVSCEKQISKIVGDMDVKSIDDGSFLLERTEFFIGNVTNAVVSQVMLVVRERNLQLIRNIPTEVKSMAVYGDQIRLQQVLAEFLLSIVRYAPMEGSVELHLCPTLNQMADGFSAVRLEFRMACAGEGVPPEKVQDMFHSSRWTSPEGLGLSVCRKILKLMNGGVQYIREFERSYFLIVIELPVPLMMMMPSS BB7CFE19C50ACBAB 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 phyc_arath Phytochrome C PHYC At5g35840 MIK22.15 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSSNTSRSCSTRSRQNSRVSSQVLVDAKLHGNFEESERLFDYSASINLNMPSSSCEIPSSAVSTYLQKIQRGMLIQPFGCLIVVDEKNLKVIAFSENTQEMLGLIPHTVPSMEQREALTIGTDVKSLFLSPGCSALEKAVDFGEISILNPITLHCRSSSKPFYAILHRIEEGLVIDLEPVSPDEVPVTAAGALRSYKLAAKSISRLQALPSGNMLLLCDALVKEVSELTGYDRVMVYKFHEDGHGEVIAECCREDMEPYLGLHYSATDIPQASRFLFMRNKVRMICDCSAVPVKVVQDKSLSQPISLSGSTLRAPHGCHAQYMSNMGSVASLVMSVTINGSDSDEMNRDLQTGRHLWGLVVCHHASPRFVPFPLRYACEFLTQVFGVQINKEAESAVLLKEKRILQTQSVLCDMLFRNAPIGIVTQSPNIMDLVKCDGAALYYRDNLWSLGVTPTETQIRDLIDWVLKSHGGNTGFTTESLMESGYPDASVLGESICGMAAVYISEKDFLFWFRSSTAKQIKWGGARHDPNDRDGKRMHPRSSFKAFMEIVRWKSVPWDDMEMDAINSLQLIIKGSLQEEHSKTVVDVPLVDNRVQKVDELCVIVNEMVRLIDTAAVPIFAVDASGVINGWNSKAAEVTGLAVEQAIGKPVSDLVEDDSVETVKNMLALALEGSEERGAEIRIRAFGPKRKSSPVELVVNTCCSRDMTNNVLGVCFIGQDVTGQKTLTENYSRVKGDYARIMWSPSTLIPPIFITNENGVCSEWNNAMQKLSGIKREEVVNKILLGEVFTTDDYGCCLKDHDTLTKLRIGFNAVISGQKNIEKLLFGFYHRDGSFIEALLSANKRTDIEGKVTGVLCFLQVPSPELQYALQVQQISEHAIACALNKLAYLRHEVKDPEKAISFLQDLLHSSGLSEDQKRLLRTSVLCREQLAKVISDSDIEGIEEGYVELDCSEFGLQESLEAVVKQVMELSIERKVQISCDYPQEVSSMRLYGDNLRLQQILSETLLSSIRFTPALRGLCVSFKVIARIEAIGKRMKRVELEFRIIHPAPGLPEDLVREMFQPLRKGTSREGLGLHITQKLVKLMERGTLRYLRESEMSAFVILTEFPLI 58FE4645501135CA phya-3 2 9 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 9 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 physical association physical association Figure 1 assay results indicate that the following binding interactions are mediated by sequences in both the phytochrome C550 and C200 clones: A/A, D/D, B/D, B/E, and D/E. phyd-1 2 8 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 8 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 18 20 physical association physical association Figure 1 assay results indicate that the following binding interactions are mediated by sequences in both the phytochrome C550 and C200 clones: A/A, D/D, B/D, B/E, and D/E. phyb-phyd-2 2 8 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 5 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 25 23 physical association physical association Figure 1 assay results indicate that the following binding interactions are mediated by sequences in both the phytochrome C550 and C200 clones: A/A, D/D, B/D, B/E, and D/E. phyb-phye-2 2 5 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 7 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Figure 1 assay results indicate that the following binding interactions are mediated by sequences in both the phytochrome C550 and C200 clones: A/A, D/D, B/D, B/E, and D/E. phyd-phye-1 2 7 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 8 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 physical association physical association Figure 1 assay results indicate that the following binding interactions are mediated by sequences in both the phytochrome C550 and C200 clones: A/A, D/D, B/D, B/E, and D/E. phyb-2 2 5 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 5 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 physical association physical association Figure 1 The b-galactosidase enzyme values for homodimerization of phyB are low and are seen only with the C200 region. Nevertheless, these assays are consistent with all of the heterodimerization interactions previously observed phyd-phyb-1 2 5 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 8 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 45 43 physical association physical association Figure 1 phye-phyd-1 2 7 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 8 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 physical association physical association Figure 1 phyb-3 1 5 unspecified role unspecified role neutral component neutral component region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction Figure S1 To further investigate this, full-length phyB, phyC, and phyE proteins were synthesized in vitro and their quaternary structures were assayed by nondenaturing gel electrophoresis. Supplemental Figure 1 online shows that, although phyB migrates as a homodimer in the presence or absence of phycocyanobilin chromophore phyc-phyb 3 10 unspecified role unspecified role bait bait 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 5 unspecified role unspecified role prey prey physical association physical association Figure 2 Figure 3 Immunoprecipitation (IP) of the extracts with anti-myc antibody pulls down either m6-phyC or phyC-m6 but fails to co-IP native phyC (Figure 2C). The tagged phyC proteins also do not pull down phyA or phyE. However, they both co-IP phyB and very weakly pull down phyD phyc-phyd 3 10 unspecified role unspecified role bait bait 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 8 unspecified role unspecified role prey prey physical association physical association Figure 2 Figure 3 Immunoprecipitation (IP) of the extracts with anti-myc antibody pulls down either m6-phyC or phyC-m6 but fails to co-IP native phyC (Figure 2C). The tagged phyC proteins also do not pull down phyA or phyE. However, they both co-IP phyB and very weakly pull down phyD The authors noted that PhyC "very weakly pull down phyD". phyc-phyb-1 3 10 unspecified role unspecified role bait bait c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 5 unspecified role unspecified role prey prey physical association physical association Figure 2 Immunoprecipitation (IP) of the extracts with anti-myc antibody pulls down either m6-phyC or phyC-m6 but fails to co-IP native phyC (Figure 2C). The tagged phyC proteins also do not pull down phyA or phyE. However, they both co-IP phyB and very weakly pull down phyD phyc-phyd-1 3 8 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 2 Immunoprecipitation (IP) of the extracts with anti-myc antibody pulls down either m6-phyC or phyC-m6 but fails to co-IP native phyC (Figure 2C). The tagged phyC proteins also do not pull down phyA or phyE. However, they both co-IP phyB and very weakly pull down phyD The authors noted that PhyC "very weakly pull down phyD". phye-phyb 3 5 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 4 Figure 7 Figure 4B shows that IP of phyE-m6 from extracts of dark-grown WT (PHYE-m6) seedlings fails to coprecipitate native phyE but does co-IP phyB and phyD. Co-IP of phyB and/or phyD with phyE-m6 is also observed in the complemented phyE(PHYE-m6) and phyBE(PHYE-m6) lines (Figure 4B). Similar results to these were obtained from IP of extracts of light-grown seedlings. phye-phyd 3 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait n-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 4 Figure 7 Figure 4B shows that IP of phyE-m6 from extracts of dark-grown WT (PHYE-m6) seedlings fails to coprecipitate native phyE but does co-IP phyB and phyD. Co-IP of phyB and/or phyD with phyE-m6 is also observed in the complemented phyE(PHYE-m6) and phyBE(PHYE-m6) lines (Figure 4B). Similar results to these were obtained from IP of extracts of light-grown seedlings. Therefore phyb-phyc-phyd-phye 3 10 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait 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 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey association association Figure 5 In Figure 5, samples from two independent dark-grown seedling anti-myc IPs were separated by electrophoresis along with standard curves of purified apoproteins, and the quantities of each immunoprecipitated phytochrome were determined from immunoblots. In dark-grown phyB(myc-PHYB) seedlings, the levels of the myc6-phyB, phyC, phyD, and phyE proteins per 1000 mg of total extract protein were calculated as 60 to 80 ng, 3 to 5 ng, 3 to 4 ng, and 0.5 to 2 ng, respectively pif3-phya-phyb-phyc 4 8 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 9 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey physical association physical association Figure 10 R/FR-Reversible Co-IP of Phytochromes with PIF3-myc6 from Seedling Extracts. (A) Rapid binding of phyA, phyB, phyC, and phyD to PIF3-m6 following a pulse of R light. The authors state that "By 1 min after the R pulse, phyA, phyB, phyC, and phyD are all detected in the PIF3-m6 immunoprecipitate, whereas they are not detected in control samples from untransformed lines or from the non-R-treated samples". Also they conclude the induction by R of co-IP of phyA, phyB, phyC, and phyD with PIF3-m6 is at least partially reversible with FR". Red light treatment as described by the authors. At least partially Far red light treatment as described by the authors. The authors noted that PIF3 interaction with PhyE "data obtained up to this time for phyE interaction with PIF3-m6 in vivo are inconclusive".