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/ Alternative complex formation of the Ca-regulated protein kinase CIPK1 controls abscisic acid-dependent and independent stress responses in Arabidopsis. Alternative complex formation of the Ca-regulated protein kinase CIPK1 controls abscisic acid-dependent and independent stress responses in Arabidopsis. Plant J. (0960-7412) 2006 imex curation D'Angelo C., Weinl S., Batistic O., Pandey GK., Cheong YH., Schultke S., Albrecht V., Ehlert B., Schulz B., Harter K., Luan S., Bock R., Kudla J. The authors used the Sos Recruitment System (SRS) in yeast to study the interaction between CBL1 and CIPK1 (see Aronheim et al (PMID: 9154808) for full details). Briefly,a bait protein is fused to a modified version of human hSos, which represents a functional homolog of the yeast Ras guanyl nucleotide-exchange factor CDC25. The mutant form of the hSos protein, lacking the C-terminal CAAX-box motif, does not localize to the plasma membrane and thus is functionally inactive. A prey protein potentially interacting with the bait can be expressed from any other plasmid harbouring a similarly strong promoter as the bait�hSos fusion. When protein�protein interaction occurs and the prey protein mediates plasma membrane localization, recruitment of the human hSos protein to the plasma membrane restores the Ras pathway, allowing growth at 35�C of the thermosensitive cdc25-2 yeast mutant (Aronheim et al., 1997). jkudla@uni-muenster.de 03-Mar-2009: Contacted by JYOTI. 03-Feb-2009: Contacted by JYOTI. orchard Only protein-protein interactions nicbe Nicotiana benthamiana bifc bimolecular fluorescence complementation bifc tag fluorescence tag visualisation by fluorescence tag fluorescence D'Angelo C., Weinl S., Batistic O., Pandey GK., Cheong YH., Schultke S., Albrecht V., Ehlert B., Schulz B., Harter K., Luan S., Bock R., Kudla J. The authors used the Sos Recruitment System (SRS) in yeast to study the interaction between CBL1 and CIPK1 (see Aronheim et al (PMID: 9154808) for full details). Briefly,a bait protein is fused to a modified version of human hSos, which represents a functional homolog of the yeast Ras guanyl nucleotide-exchange factor CDC25. The mutant form of the hSos protein, lacking the C-terminal CAAX-box motif, does not localize to the plasma membrane and thus is functionally inactive. A prey protein potentially interacting with the bait can be expressed from any other plasmid harbouring a similarly strong promoter as the bait�hSos fusion. When protein�protein interaction occurs and the prey protein mediates plasma membrane localization, recruitment of the human hSos protein to the plasma membrane restores the Ras pathway, allowing growth at 35�C of the thermosensitive cdc25-2 yeast mutant (Aronheim et al., 1997). jkudla@uni-muenster.de Accepted 2009-FEB-19 by ORCHARD. 03-Mar-2009: Contacted by JYOTI. Plant J. (0960-7412) 2006 orchard Only protein-protein interactions imex curation Alternative complex formation of the Ca-regulated protein kinase CIPK1 controls abscisic acid-dependent and independent stress responses in Arabidopsis. Alternative complex formation of the Ca-regulated protein kinase CIPK1 controls abscisic acid-dependent and independent stress responses in Arabidopsis. Plant J. (0960-7412) 2006 imex curation D'Angelo C., Weinl S., Batistic O., Pandey GK., Cheong YH., Schultke S., Albrecht V., Ehlert B., Schulz B., Harter K., Luan S., Bock R., Kudla J. The authors used the Sos Recruitment System (SRS) in yeast to study the interaction between CBL1 and CIPK1 (see Aronheim et al (PMID: 9154808) for full details). Briefly,a bait protein is fused to a modified version of human hSos, which represents a functional homolog of the yeast Ras guanyl nucleotide-exchange factor CDC25. The mutant form of the hSos protein, lacking the C-terminal CAAX-box motif, does not localize to the plasma membrane and thus is functionally inactive. A prey protein potentially interacting with the bait can be expressed from any other plasmid harbouring a similarly strong promoter as the bait�hSos fusion. When protein�protein interaction occurs and the prey protein mediates plasma membrane localization, recruitment of the human hSos protein to the plasma membrane restores the Ras pathway, allowing growth at 35�C of the thermosensitive cdc25-2 yeast mutant (Aronheim et al., 1997). jkudla@uni-muenster.de 03-Mar-2009: Contacted by JYOTI. 03-Feb-2009: Contacted by JYOTI. orchard Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast reverse rrs reverse ras recruitment system reverse RRS reverse rrs predetermined predetermined participant predetermined D'Angelo C., Weinl S., Batistic O., Pandey GK., Cheong YH., Schultke S., Albrecht V., Ehlert B., Schulz B., Harter K., Luan S., Bock R., Kudla J. The authors used the Sos Recruitment System (SRS) in yeast to study the interaction between CBL1 and CIPK1 (see Aronheim et al (PMID: 9154808) for full details). Briefly,a bait protein is fused to a modified version of human hSos, which represents a functional homolog of the yeast Ras guanyl nucleotide-exchange factor CDC25. The mutant form of the hSos protein, lacking the C-terminal CAAX-box motif, does not localize to the plasma membrane and thus is functionally inactive. A prey protein potentially interacting with the bait can be expressed from any other plasmid harbouring a similarly strong promoter as the bait�hSos fusion. When protein�protein interaction occurs and the prey protein mediates plasma membrane localization, recruitment of the human hSos protein to the plasma membrane restores the Ras pathway, allowing growth at 35�C of the thermosensitive cdc25-2 yeast mutant (Aronheim et al., 1997). jkudla@uni-muenster.de Accepted 2009-FEB-19 by ORCHARD. 03-Mar-2009: Contacted by JYOTI. Plant J. (0960-7412) 2006 orchard Only protein-protein interactions imex curation 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 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 cnbl9_arath Calcineurin B-like protein 9 CBL9 At5g47100 K14A3.5 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGCFHSTAAREFPDHENPVKLASETAFSVSEVEALYELFKSISSSVVDDGLINKEEFQLALFKNRKKENLFANRIFDLFDVKRKGVIDFGDFVRSLNVFHPNASLEEKTDFTFRLYDMDCTGFIERQEVKQMLIALLCESEMKLADDTIEMILDQTFEDADVDRDGKIDKTEWSNFVIKNPSLLKIMTLPYLRDITTTFPSFVFNSEVDEIAT E848102CD14211EE cipk1-cbl1-2 2 4 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 3 unspecified role unspecified role prey prey physical association physical association Complementation of the yeast cdc25-2 mutant through interaction of CBL1 with CIPK1 at the plasma membrane. Figure 2 Details of the NAF domain were taken from Albrecht et al (PMID: 11230129) as referenced in the text. The authors concluded the myristoylation of the interacting CBL protein is required for targeting CIPK1 to the plasma membrane". cipk1-cbl1-3 1 4 unspecified role unspecified role neutral component neutral component region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region N-terminal YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 3 unspecified role unspecified role neutral component neutral component region C-terminal YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association To directly establish the physical interaction of CBL1 and CBL9 with CIPK1 in planta, we used bimolecular fluorescence complementation (BiFC) in N. benthamiana protoplasts (Walter et al., 2004). In these assays, both CBL1/CIPK1 and CBL9/CIPK1 complexes were observed exclusively at the plasma membrane Figure 3 Details of the NAF domain were taken from Albrecht et al (PMID: 11230129) as referenced in the text. The authors concluded the "data provide compelling evidence for CBL1 and CBL9 recruiting CIPK1 to the plasma membrane by interaction via the NAF domain. cipk1-cbl9 1 5 unspecified role unspecified role neutral component neutral component region C-terminal YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role neutral component neutral component region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region N-terminal YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association To directly establish the physical interaction of CBL1 and CBL9 with CIPK1 in planta, we used bimolecular fluorescence complementation (BiFC) in N. benthamiana protoplasts (Walter et al., 2004). In these assays, both CBL1/CIPK1 and CBL9/CIPK1 complexes were observed exclusively at the plasma membrane Figure 3 Details of the NAF domain were taken from Albrecht et al (PMID: 11230129) as referenced in the text. The authors concluded the "data provide compelling evidence for CBL1 and CBL9 recruiting CIPK1 to the plasma membrane by interaction via the NAF domain.