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/ BRI1/BAK1, a receptor kinase pair mediating brassinosteroid signaling. BRI1/BAK1, a receptor kinase pair mediating brassinosteroid signaling. Cell (0092-8674) 2002 Nam KH., Li J. Arabidopsis cDNA library jian@umich.edu; 14-Nov-2008: Contacted by JYOTI. orchard yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Nam KH., Li J. Arabidopsis cDNA library jian@umich.edu; Accepted 2008-SEP-26 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Cell (0092-8674) 2002 orchard BRI1/BAK1, a receptor kinase pair mediating brassinosteroid signaling. BRI1/BAK1, a receptor kinase pair mediating brassinosteroid signaling. Cell (0092-8674) 2002 Nam KH., Li J. Arabidopsis cDNA library jian@umich.edu; 14-Nov-2008: Contacted by JYOTI. orchard 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 Nam KH., Li J. Note: it is not clear from the analysis of kinase activity presented in Fig 2E , if transphosphorylation or autophosphorylation is occuring and because of this ambiguity the data was not curated. Transgenic Arabidopsis plants expressing epitope tagged proteins were used in this study. Arabidopsis cDNA library jian@umich.edu; Accepted 2008-SEP-26 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Cell (0092-8674) 2002 orchard BRI1/BAK1, a receptor kinase pair mediating brassinosteroid signaling. BRI1/BAK1, a receptor kinase pair mediating brassinosteroid signaling. Cell (0092-8674) 2002 Nam KH., Li J. Arabidopsis cDNA library jian@umich.edu; 14-Nov-2008: Contacted by JYOTI. orchard 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 Nam KH., Li J. Note: it is not clear from the analysis of kinase activity presented in Fig 2E , if transphosphorylation or autophosphorylation is occuring and because of this ambiguity the data was not curated. Transgenic Arabidopsis plants expressing epitope tagged proteins were used in this study. Arabidopsis cDNA library jian@umich.edu; Accepted 2008-SEP-26 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Cell (0092-8674) 2002 orchard BRI1/BAK1, a receptor kinase pair mediating brassinosteroid signaling. BRI1/BAK1, a receptor kinase pair mediating brassinosteroid signaling. Cell (0092-8674) 2002 Nam KH., Li J. Arabidopsis cDNA library jian@umich.edu; 14-Nov-2008: Contacted by JYOTI. orchard 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Nam KH., Li J. Arabidopsis cDNA library jian@umich.edu; Accepted 2008-SEP-26 by ORCHARD. Cell (0092-8674) 2002 orchard BRI1/BAK1, a receptor kinase pair mediating brassinosteroid signaling. BRI1/BAK1, a receptor kinase pair mediating brassinosteroid signaling. Cell (0092-8674) 2002 Nam KH., Li J. Arabidopsis cDNA library jian@umich.edu; 14-Nov-2008: Contacted by JYOTI. orchard arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Nam KH., Li J. Note: it is not clear from the analysis of kinase activity presented in Fig 2E , if transphosphorylation or autophosphorylation is occuring and because of this ambiguity the data was not curated. Transgenic Arabidopsis plants expressing epitope tagged proteins were used in this study. Arabidopsis cDNA library jian@umich.edu; Accepted 2008-SEP-26 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Cell (0092-8674) 2002 orchard bak1_arath BRASSINOSTEROID INSENSITIVE 1-associated receptor kinase 1 BAK1 SERK3 At4g33430 F17M5.190 Somatic embryogenesis receptor-like kinase 3 ELG Somatic embryogenesis receptor kinase 3 Protein ELONGATED protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MERRLMIPCFFWLILVLDLVLRVSGNAEGDALSALKNSLADPNKVLQSWDATLVTPCTWFHVTCNSDNSVTRVDLGNANLSGQLVMQLGQLPNLQYLELYSNNITGTIPEQLGNLTELVSLDLYLNNLSGPIPSTLGRLKKLRFLRLNNNSLSGEIPRSLTAVLTLQVLDLSNNPLTGDIPVNGSFSLFTPISFANTKLTPLPASPPPPISPTPPSPAGSNRITGAIAGGVAAGAALLFAVPAIALAWWRRKKPQDHFFDVPAEEDPEVHLGQLKRFSLRELQVASDNFSNKNILGRGGFGKVYKGRLADGTLVAVKRLKEERTQGGELQFQTEVEMISMAVHRNLLRLRGFCMTPTERLLVYPYMANGSVASCLRERPESQPPLDWPKRQRIALGSARGLAYLHDHCDPKIIHRDVKAANILLDEEFEAVVGDFGLAKLMDYKDTHVTTAVRGTIGHIAPEYLSTGKSSEKTDVFGYGVMLLELITGQRAFDLARLANDDDVMLLDWVKGLLKEKKLEALVDVDLQGNYKDEEVEQLIQVALLCTQSSPMERPKMSEVVRMLEGDGLAERWEEWQKEEMFRQDFNYPTHHPAVSGWIIGDSTSQIENEYPSGPR DIP protein Q94F62 original sequence version: 119 DIP protein Q94F62 original sequence version: 129 DIP protein Q94F62 original sequence version: 94 DIP protein Q94F62 original sequence version: 100 DIP protein Q94F62 original sequence version: 139 DIP protein Q94F62 original sequence version: 121 DIP protein Q94F62 original sequence version: 141 DIP protein Q94F62 original sequence version: 113 547B3F38FB46E1DD bri1_arath Protein BRASSINOSTEROID INSENSITIVE 1 Brassinosteroid LRR receptor kinase BRI1 F23K16.30 At4g39400 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MKTFSSFFLSVTTLFFFSFFSLSFQASPSQSLYREIHQLISFKDVLPDKNLLPDWSSNKNPCTFDGVTCRDDKVTSIDLSSKPLNVGFSAVSSSLLSLTGLESLFLSNSHINGSVSGFKCSASLTSLDLSRNSLSGPVTTLTSLGSCSGLKFLNVSSNTLDFPGKVSGGLKLNSLEVLDLSANSISGANVVGWVLSDGCGELKHLAISGNKISGDVDVSRCVNLEFLDVSSNNFSTGIPFLGDCSALQHLDISGNKLSGDFSRAISTCTELKLLNISSNQFVGPIPPLPLKSLQYLSLAENKFTGEIPDFLSGACDTLTGLDLSGNHFYGAVPPFFGSCSLLESLALSSNNFSGELPMDTLLKMRGLKVLDLSFNEFSGELPESLTNLSASLLTLDLSSNNFSGPILPNLCQNPKNTLQELYLQNNGFTGKIPPTLSNCSELVSLHLSFNYLSGTIPSSLGSLSKLRDLKLWLNMLEGEIPQELMYVKTLETLILDFNDLTGEIPSGLSNCTNLNWISLSNNRLTGEIPKWIGRLENLAILKLSNNSFSGNIPAELGDCRSLIWLDLNTNLFNGTIPAAMFKQSGKIAANFIAGKRYVYIKNDGMKKECHGAGNLLEFQGIRSEQLNRLSTRNPCNITSRVYGGHTSPTFDNNGSMMFLDMSYNMLSGYIPKEIGSMPYLFILNLGHNDISGSIPDEVGDLRGLNILDLSSNKLDGRIPQAMSALTMLTEIDLSNNNLSGPIPEMGQFETFPPAKFLNNPGLCGYPLPRCDPSNADGYAHHQRSHGRRPASLAGSVAMGLLFSFVCIFGLILVGREMRKRRRKKEAELEMYAEGHGNSGDRTANNTNWKLTGVKEALSINLAAFEKPLRKLTFADLLQATNGFHNDSLIGSGGFGDVYKAILKDGSAVAIKKLIHVSGQGDREFMAEMETIGKIKHRNLVPLLGYCKVGDERLLVYEFMKYGSLEDVLHDPKKAGVKLNWSTRRKIAIGSARGLAFLHHNCSPHIIHRDMKSSNVLLDENLEARVSDFGMARLMSAMDTHLSVSTLAGTPGYVPPEYYQSFRCSTKGDVYSYGVVLLELLTGKRPTDSPDFGDNNLVGWVKQHAKLRISDVFDPELMKEDPALEIELLQHLKVAVACLDDRAWRRPTMVQVMAMFKEIQAGSGIDSQSTIRSIEDGGFSTIEMVDMSIKEVPEGKL DIP protein O22476 original sequence version: 104 DIP protein O22476 original sequence version: 120 DIP protein O22476 original sequence version: 146 DIP protein O22476 original sequence version: 130 DIP protein O22476 original sequence version: 123 C7FBA1C21294E600 brl1-bak1-5 4 7 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind range range range range 6 unspecified role unspecified role prey prey physical association physical association Based on previous genetic evidence that the cytoplasmic kinase domain of BRI1 (BRI1CK) is crucial for BR signaling (Li and Chory 1997; Noguchi et al. 1999 and Friedrichsen, D.M., Joazeiro, C.A., Li, J., Hunter, T. and Chory, J., 2000. Brassinosteroid-insensitive-1 is a ubiquitously expressed leucine-rich repeat receptor serine/threonine kinase. Plant Physiol. 123, pp. 1247�1256. Full Text via CrossRefFriedrichsen et al. 2000), we used this domain as bait and conducted a yeast two-hybrid screen for potential BRI1-interacting proteins. Among the 96 BRI1-interacting clones, there are six overlapping cDNAs encoding an LRR-containing receptor-like kinase of 615 amino acids, with their 5′-ends near the activation loop (A loop) of its catalytic domain. We named this interactor as BAK1 for BRI1 Associated receptor Kinase 1. The range of the kinase domain used in the yeast two hybrid assay was not described in the text. The range is taken from Li and Chroy (1997) , PMID PMID: 9298904. Because the range is not directly taken from the source text, it is presented as a fuzzy range to indicate that the range was not explicitly stated by the authors of the paper under curation. Note that the authors state that "all the recovered BAK1 clones are partial cDNAs encoding the C-terminal part of the BAK1 protein". However the range of the cDNA's recovered is not given. brl1-bak1-6 1 7 unspecified role unspecified role bait bait c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 2D This result indicates that the full-length BRI1 interacted with the full-length BAK1 in yeast. brl1-bak1-7 3 6 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role bait bait region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 3D BAK1 Directly Interacts with BRI1 in Plants brl1-bak1-8 2 6 unspecified role unspecified role bait bait region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey physical association physical association Figure 3F BRI1 Is Capable of Interacting with BAK1 in Plants brl1-bak1-9 5 6 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role bait bait physical association physical association Figure 3E BRI1 Is Capable of Interacting with BAK1 in Plants