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/ N-terminal domain-mediated homodimerization is required for photoreceptor activity of Arabidopsis CRYPTOCHROME 1. N-terminal domain-mediated homodimerization is required for photoreceptor activity of Arabidopsis CRYPTOCHROME 1. Plant Cell (1040-4651) 2005 imex curation Sang Y., Li QH., Rubio V., Zhang YC., Mao J., Deng XW., Yang HQ. hqyang@sibs.ac.cn 22-Jan-2008: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Sang Y., Li QH., Rubio V., Zhang YC., Mao J., Deng XW., Yang HQ. Only results for GST pulldown under blue light conditions were presented by the authors. Pull down assays were undertaken under blue light and darkness. hqyang@sibs.ac.cn Accepted 2007-DEC-21 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. Plant Cell (1040-4651) 2005 orchard Only protein-protein interactions imex curation N-terminal domain-mediated homodimerization is required for photoreceptor activity of Arabidopsis CRYPTOCHROME 1. N-terminal domain-mediated homodimerization is required for photoreceptor activity of Arabidopsis CRYPTOCHROME 1. Plant Cell (1040-4651) 2005 imex curation Sang Y., Li QH., Rubio V., Zhang YC., Mao J., Deng XW., Yang HQ. hqyang@sibs.ac.cn 22-Jan-2008: Contacted by JYOTI. orchard Only protein-protein interactions arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress crosslink cross-linking study crosslink anti tag western anti tag western blot anti tag western Sang Y., Li QH., Rubio V., Zhang YC., Mao J., Deng XW., Yang HQ. Transgenic Arabidopsis (Columbia ecotype) expressing an epitope tagged Cryptochrome 1 protein were used in this study. The seeds were were put in a 48C refrigerator for 3 to 5 d, induced for germination in white light for 24 h, and finally transferred to the dark and continuous blue light (20 micro mol per second per meter squared.) Seedlings were cross-linked with formaldehyde treatment. hqyang@sibs.ac.cn Accepted 2007-DEC-21 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. Plant Cell (1040-4651) 2005 orchard Only protein-protein interactions imex curation N-terminal domain-mediated homodimerization is required for photoreceptor activity of Arabidopsis CRYPTOCHROME 1. N-terminal domain-mediated homodimerization is required for photoreceptor activity of Arabidopsis CRYPTOCHROME 1. Plant Cell (1040-4651) 2005 imex curation Sang Y., Li QH., Rubio V., Zhang YC., Mao J., Deng XW., Yang HQ. hqyang@sibs.ac.cn 22-Jan-2008: Contacted by JYOTI. orchard Only protein-protein interactions arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Sang Y., Li QH., Rubio V., Zhang YC., Mao J., Deng XW., Yang HQ. Transgenic Arabidopsis (Columbia ecotype) expressing an epitope tagged Cryptochrome 1 protein were used in this study. Seedlings were grown for 4 d under continuous blue light (5 micro mol per second per meter squared) continuous darkness. hqyang@sibs.ac.cn Accepted 2007-DEC-21 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. Plant Cell (1040-4651) 2005 orchard Only protein-protein interactions imex curation N-terminal domain-mediated homodimerization is required for photoreceptor activity of Arabidopsis CRYPTOCHROME 1. N-terminal domain-mediated homodimerization is required for photoreceptor activity of Arabidopsis CRYPTOCHROME 1. Plant Cell (1040-4651) 2005 imex curation Sang Y., Li QH., Rubio V., Zhang YC., Mao J., Deng XW., Yang HQ. hqyang@sibs.ac.cn 22-Jan-2008: 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 Sang Y., Li QH., Rubio V., Zhang YC., Mao J., Deng XW., Yang HQ. Transgenic Arabidopsis (Columbia ecotype) expressing an epitope tagged Cryptochrome 1 protein were used in this study. Seedlings were grown for for 4 days in continuous darkness prior to assay. hqyang@sibs.ac.cn Accepted 2007-DEC-21 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. Plant Cell (1040-4651) 2005 orchard Only protein-protein interactions imex curation N-terminal domain-mediated homodimerization is required for photoreceptor activity of Arabidopsis CRYPTOCHROME 1. N-terminal domain-mediated homodimerization is required for photoreceptor activity of Arabidopsis CRYPTOCHROME 1. Plant Cell (1040-4651) 2005 imex curation Sang Y., Li QH., Rubio V., Zhang YC., Mao J., Deng XW., Yang HQ. hqyang@sibs.ac.cn 22-Jan-2008: 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 Sang Y., Li QH., Rubio V., Zhang YC., Mao J., Deng XW., Yang HQ. hqyang@sibs.ac.cn Accepted 2007-DEC-21 by ORCHARD. Plant Cell (1040-4651) 2005 orchard Only protein-protein interactions imex curation cry2_arath Cryptochrome-2 Blue light photoreceptor CRY2 PHH1 Protein PHR homolog 1 Protein SUPPRESSOR OF elf3 20 SEL20 F19P19.14 At1g04400 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQFYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLYDPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSIESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLNEFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGEESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPLVDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQYISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKASGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAAPDEIVADSFEALGANTIKEPGLCPSVSSNDQQVPSAVRYNGSKRVKPEEEEERDMKKSRGFDERELFSTAESSSSSSVFFVSQSCSLASEGKNLEGIQDSSDQITTSLGKNGCK DIP protein Q96524 original sequence version: 123 DIP protein Q96524 original sequence version: 101 DIP protein Q96524 original sequence version: 110 082E311301465904 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 cry1-cry1 5 7 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 7 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 10 12 physical association physical association 2 Interaction of CNT with Full-Length CRY and CNT Itself in Yeast Cells. cry2-cry2 5 6 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 6 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 17 15 physical association physical association 2 Interaction of CNT with Full-Length CRY and CNT Itself in Yeast Cells. cry1-cry1-1 1 7 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 region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 7 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast direct interaction direct interaction 3 Pulldown assay using these proteins showed that both CRY and CNT bound to CRY cry2-cry2-1 1 6 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 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 in vitro In vitro 6 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined schpm Schizosaccharomyces pombe (fission yeast) Fission yeast direct interaction direct interaction 3 Pulldown assay using these proteins showed that both CRY and CNT bound to CRY cry1-cry1-2 2 7 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q43125:p.Gly347Arg mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain G R Wed May 02 20:23:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly347arg' Tue Jun 30 17:41:22 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly347Arg' 7 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q43125:p.Gly347Arg mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain G R Wed May 02 15:45:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly347arg' Tue Jun 30 19:14:19 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly347Arg' physical association physical association 4 After treatment with formaldehyde, a novel high molecular mass band, approximately twice as high as that of the Myc-CRY1 monomer, recognized by anti-Myc antibody, was observed after SDS-PAGE separation of the protein extracts prepared from dark-grown and blue light�grown, and blue light�irradiated seedlings expressing Myc-CRY1 cry1-cry1-3 3 7 unspecified role unspecified role neutral component neutral component region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role neutral component neutral component region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 4 To further confirm the homodimerization of CRY1 in vivo, we generated transgenic plants expressing an N-terminal fusion of the TAPa tag to the CRY1 protein (TAPa-CRY1) (Saijo et al., 2003; Rubio et al., 2005) in the wild-type background and conducted gel filtration studies. In both dark and blue light conditions, approximately equal amounts of dimer and monomer of the native CRY1 were obtained (Figure 4C), whereas the TAPa-CRY1 was found almost all as a dimer. cry1-cry1-4 4 7 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey physical association physical association 4 Furthermore, coimmunoprecipitation assays showed that TAPa-CRY1 bound endogenous CRY1, whereas a TAPa�green fluorescent protein (GFP) fusion did not (Figures 4D and 4E). Therefore, these data indicate that CRY1 homodimerizes in vivo in a light-independent manner. cry1-cry1-5 5 7 unspecified role unspecified role bait bait Q43125:p.Ala462Val mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain A V Wed May 02 15:04:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala462val' Tue Jun 30 17:25:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala462Val' n-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain Q43125:p.Gly347Arg mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain G R Wed May 02 17:07:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly347arg' Tue Jun 30 19:27:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly347Arg' Q43125:p.Ser66Asp mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain S D Wed May 02 15:29:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser66asp' Tue Jun 30 16:37:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser66Asp' 7 unspecified role unspecified role prey prey physical association physical association 5 Yeast two-hybrid assays showed that all of these point mutations abolished CNT1�CRY1 interaction