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/ Novel CIPK1-associated proteins in Arabidopsis contain an evolutionarily conserved C-terminal region that mediates nuclear localization. Novel CIPK1-associated proteins in Arabidopsis contain an evolutionarily conserved C-terminal region that mediates nuclear localization. Plant Physiol 2005 imex curation Ok SH, Jeong HJ, Bae JM, Shin JS, Luan S, Kim KN. knkim@sejong.ac.kr 27-Oct-2007: Contacted by JYOTI. Arabidopsis cDNA library 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Ok SH, Jeong HJ, Bae JM, Shin JS, Luan S, Kim KN. knkim@sejong.ac.kr Accepted 2009-OCT-1 by ORCHARD. Arabidopsis cDNA library Plant Physiol 2005 orchard Only protein-protein interactions imex curation Novel CIPK1-associated proteins in Arabidopsis contain an evolutionarily conserved C-terminal region that mediates nuclear localization. Novel CIPK1-associated proteins in Arabidopsis contain an evolutionarily conserved C-terminal region that mediates nuclear localization. Plant Physiol 2005 imex curation Ok SH, Jeong HJ, Bae JM, Shin JS, Luan S, Kim KN. knkim@sejong.ac.kr 27-Oct-2007: Contacted by JYOTI. Arabidopsis cDNA library 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 Ok SH, Jeong HJ, Bae JM, Shin JS, Luan S, Kim KN. knkim@sejong.ac.kr Accepted 2009-OCT-1 by ORCHARD. 27-Oct-2007: Contacted by JYOTI. Arabidopsis cDNA library Plant Physiol 2005 orchard Only protein-protein interactions imex curation Novel CIPK1-associated proteins in Arabidopsis contain an evolutionarily conserved C-terminal region that mediates nuclear localization. Novel CIPK1-associated proteins in Arabidopsis contain an evolutionarily conserved C-terminal region that mediates nuclear localization. Plant Physiol 2005 imex curation Ok SH, Jeong HJ, Bae JM, Shin JS, Luan S, Kim KN. knkim@sejong.ac.kr 27-Oct-2007: Contacted by JYOTI. Arabidopsis cDNA library orchard Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Ok SH, Jeong HJ, Bae JM, Shin JS, Luan S, Kim KN. knkim@sejong.ac.kr Accepted 2009-OCT-1 by ORCHARD. 27-Oct-2007: Contacted by JYOTI. Arabidopsis cDNA library Plant Physiol 2005 orchard Only protein-protein interactions imex curation cipk1_arath CBL-interacting serine/threonine-protein kinase 1 CIPK1 PKS13 SnRK3.16 MKP6.6 At3g17510 SOS2-like protein kinase PKS13 SNF1-related kinase 3.16 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MVRRQEEEKKAEKGMRLGKYELGRTLGEGNFGKVKFAKDTVSGHSFAVKIIDKSRIADLNFSLQIKREIRTLKMLKHPHIVRLHEVLASKTKINMVMELVTGGELFDRIVSNGKLTETDGRKMFQQLIDGISYCHSKGVFHRDLKLENVLLDAKGHIKITDFGLSALPQHFRDDGLLHTTCGSPNYVAPEVLANRGYDGAASDIWSCGVILYVILTGCLPFDDRNLAVLYQKICKGDPPIPRWLSPGARTMIKRMLDPNPVTRITVVGIKASEWFKLEYIPSIPDDDDEEEVDTDDDAFSIQELGSEEGKGSDSPTIINAFQLIGMSSFLDLSGFFEQENVSERRIRFTSNSSAKDLLEKIETAVTEMGFSVQKKHAKLRVKQEERNQKGQVGLSVTAEVFEIKPSLNVVELRKSYGDSCLYRQLYERLLKDVGTSSPEQEIVT 63DB8AF69CCAA5C9 cnbl3_arath Calcineurin B-like protein 3 SOS3-like calcium-binding protein 6 CBL3 SCABP6 At4g26570 T15N24.20 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSQCIDGFKHVCSSFFRCFDIDIYKQSGGLGDPELLARETVFSVSEIEALYELFKKISSAVIDDGLINKEEFQLALFKTNKKESLFADRVFDLFDTKHNGILGFEEFARALSVFHPNAPIEDKIDFSFQLYDLKQQGFIERQEVKQMVVATLAESGMNLSDEIIESIIDKTFEEADTKHDGRIDKEEWRTLVLRHPSLLKNMTLQYLKDITTTFPSFVFHSQVEDT F78222E98F64983E cnbl1_arath Calcineurin B-like protein 1 SOS3-like calcium-binding protein 5 CBL1 SCABP5 At4g17615 dl4845w FCAALL.122 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGCFHSKAAKEFRGHEDPVKLASETAFSVSEVEALFELFKSISSSVVDDGLINKEEFQLALFKSRKRENIFANRIFDMFDVKRKGVIDFGDFVRSLNVFHPNASLEDKIDFTFRLYDMDCTGYIERQEVKQMLIALLCESEMKLADETIEIILDKTFEDADVNQDGKIDKLEWSDFVNKNPSLLKIMTLPYLRDITTTFPSFVFHSEVDEIAT 1C310FC449D027B1 ect1_arath YTH domain-containing protein ECT1 ECT1 Protein EVOLUTIONARILY CONSERVED C-TERMINAL REGION 1 T11I18.6 At3g03950 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAGAASSDRLVTSFPLLDTADLFQDLSLGSDANEVPMNFTKGSFQHPYGHAPYGASSHGSERRPNMNAGNLLNGGDSIGSYPWGYIPANYPSGGYPDPRFGYDRNSNHSSFSHLMNPHSSQEVPSFDQLGYNDHLYSNHGLYGLYGNVIDSGHAYGTFGYDSWKLGRGWYPVDGYRKTRSFNHGRGYSDEKADRLNELCRGPRSSDFKNPQVLNSSMLDAMKQDVSAVDLQRYNGENFPESFVKAKFFVIKSYSEDDVHNCIKYGAWSSTPTGNKKLNAAYYEAKENSQECPVYLLFSVNASGQFVGLAEMVGPVDFNKTMEYWQQDKWIGCFPVKWHIIKDIPNSLLRHITLANNENKPVTNSRDTQEVNLEHGTKIIKIFKEYMSKTCILDDYKFYETRQKIIRDKKIKQKKQALDGASGETINLS E9BC918D9F94FDEC ect2_arath YTH domain-containing protein ECT2 ECT2 Protein EVOLUTIONARILY CONSERVED C-TERMINAL REGION 2 MRP15.12 At3g13460 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MATVAPPADQATDLLQKLSLDSPAKASEIPEPNKKTAVYQYGGVDVHGQVPSYDRSLTPMLPSDAADPSVCYVPNPYNPYQYYNVYGSGQEWTDYPAYTNPEGVDMNSGIYGENGTVVYPQGYGYAAYPYSPATSPAPQLGGEGQLYGAQQYQYPNYFPNSGPYASSVATPTQPDLSANKPAGVKTLPADSNNVASAAGITKGSNGSAPVKPTNQATLNTSSNLYGMGAPGGGLAAGYQDPRYAYEGYYAPVPWHDGSKYSDVQRPVSGSGVASSYSKSSTVPSSRNQNYRSNSHYTSVHQPSSVTGYGTAQGYYNRMYQNKLYGQYGSTGRSALGYGSSGYDSRTNGRGWAATDNKYRSWGRGNSYYYGNENNVDGLNELNRGPRAKGTKNQKGNLDDSLEVKEQTGESNVTEVGEADNTCVVPDREQYNKEDFPVDYANAMFFIIKSYSEDDVHKSIKYNVWASTPNGNKKLAAAYQEAQQKAGGCPIFLFFSVNASGQFVGLAEMTGPVDFNTNVEYWQQDKWTGSFPLKWHIVKDVPNSLLKHITLENNENKPVTNSRDTQEVKLEQGLKIVKIFKEHSSKTCILDDFSFYEVRQKTILEKKAKQTQKQVSEEKVTDEKKESATAESASKESPAAVQTSSDVKVAENGSVAKPVTGDVVANGC 30AD740C806B1044 cipk1-cbl1-4 1 6 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait physical association physical association Because CBL1 and CBL3 were previously known to associate with CIPK1 (Shi et al., 1999), isolation of these genes indicated that the yeast two-hybrid screening was successfully carried out. cipk1-cbl3-1 1 5 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait physical association physical association cipk1-ect1 1 7 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait physical association physical association cipk1-ect2 1 8 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait physical association physical association cipk1-ect1-1 2 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 4 unspecified role unspecified role bait bait n-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 25 23 physical association physical association Figure 2A Figure 3 As shown in Figure 2A, yeast that carry both pGBT.CIPK1 and pGAD.ECT1 grew well on the selection medium (SC-HLT), indicating expression of the HIS3 reporter gene. Caution should be exercised by the user since the length of the protein ECT1 in the paper differs from the length provided by the UniProt record for this protein. cipk1-ect2-1 2 4 unspecified role unspecified role bait bait n-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 8 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 28 30 physical association physical association Figure 2C Figure 3 These results indicate that the full-length ECT1 and ECT2 proteins respectively interact with CIPK1 in the yeast two-hybrid system Caution should be exercised by the user since the length of the protein ECT2 in the paper differs from the length provided by the UniProt record for this protein. cipk1-ect1-2 3 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 ecoli Escherichia coli (strain K12) 4 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 5 CIPK1 physically interacts with ECT1 in vitro. Caution should be exercised by the user since the length of the protein ECT1 in the paper differs from the length provided by the UniProt record for this protein. cipk1-ect1-3 3 7 unspecified role unspecified role prey prey 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 ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role prey prey arath-root Arabidopsis thaliana root tissue root physical association physical association Figure 6 Affinity purification of CIPK1 by GST-ECT1 from the Arabidopsis root total protein extract