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/ http://www.ebi.ac.uk/ pandey-2004-4 The Arabidopsis putative G protein-coupled receptor GCR1 interacts with the G protein alpha subunit GPA1 and regulates abscisic acid signaling. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast ub reconstruction ubiquitin reconstruction ub reconstruction predetermined predetermined participant predetermined Pandey S., Assmann SM. sma3@psu.edu Plant Cell (1040-4651) 2004 pandey-2004-3 The Arabidopsis putative G protein-coupled receptor GCR1 interacts with the G protein alpha subunit GPA1 and regulates abscisic acid signaling. 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 Pandey S., Assmann SM. gcr1-3 mutant plants were transformed with a full-length GCR1 cDNA fused with a FLAG epitope tag under the control of a glucocorticoid-inducible promoter. Protein expression was induced by treating plants with dexmethasone. sma3@psu.edu Plant Cell (1040-4651) 2004 pandey-2004-1 The Arabidopsis putative G protein-coupled receptor GCR1 interacts with the G protein alpha subunit GPA1 and regulates abscisic acid signaling. in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Pandey S., Assmann SM. sma3@psu.edu Plant Cell (1040-4651) 2004 pandey-2004-2 The Arabidopsis putative G protein-coupled receptor GCR1 interacts with the G protein alpha subunit GPA1 and regulates abscisic acid signaling. 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 Pandey S., Assmann SM. gcr1-3 mutant plants were transformed with a full-length GCR1 cDNA fused with a FLAG epitope tag under the control of a glucocorticoid-inducible promoter. Protein expression was induced by treating plants with dexmethasone. sma3@psu.edu Plant Cell (1040-4651) 2004 gpa1_arath Guanine nucleotide-binding protein alpha-1 subunit GPA1 At2g26300 T1D16.6 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGLLCSRSRHHTEDTDENTQAAEIERRIEQEAKAEKHIRKLLLLGAGESGKSTIFKQIKLLFQTGFDEGELKSYVPVIHANVYQTIKLLHDGTKEFAQNETDSAKYMLSSESIAIGEKLSEIGGRLDYPRLTKDIAEGIETLWKDPAIQETCARGNELQVPDCTKYLMENLKRLSDINYIPTKEDVLYARVRTTGVVEIQFSPVGENKKSGEVYRLFDVGGQRNERRKWIHLFEGVTAVIFCAAISEYDQTLFEDEQKNRMMETKELFDWVLKQPCFEKTSFMLFLNKFDIFEKKVLDVPLNVCEWFRDYQPVSSGKQEIEHAYEFVKKKFEELYYQNTAPDRVDRVFKIYRTTALDQKLVKKTFKLVDETLRRRNLLEAGLL gcr1_arath G-protein coupled receptor 1 GCR1 F11A17.17 At1g48270 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSAVLTAGGGLTAGDRSIITAINTGASSLSFVGSAFIVLCYCLFKELRKFSFKLVFYLALSDMLCSFFLIVGDPSKGFICYAQGYTTHFFCVASFLWTTTIAFTLHRTVVKHKTDVEDLEAMFHLYVWGTSLVVTVIRSFGNNHSHLGPWCWTQTGLKGKAVHFLTFYAPLWGAILYNGFTYFQVIRMLRNARRMAVGMSDRVDQFDNRAELKVLNRWGYYPLILIGSWAFGTINRIHDFIEPGHKIFWLSVLDVGTAALMGLFNSIAYGFNSSVRRAIHERLELFLPERLYRWLPSNFRPKNHLILHQQQQQRSEMVSLKTEDQQ gcr1-gpa1-4 Interaction detected by ubiquitin reconstruction 143213 n/a 143217 unspecified role unspecified role bait bait arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress n/a 143218 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress physical association physical association false false false Figs. 2 and 4 GPA1-Cub protein interacts with NubG-GCR1 but not with GCR1-NubG . GCR1-Cub, however, does not show interaction with GPA1-Nub constructs, suggesting that a free GCR1 C-terminus is required for interaction. gcr1-gpa1-5 Interaction detected by ubiquitin reconstruction 143213 n/a 143218 unspecified role unspecified role bait bait arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress n/a 143217 unspecified role unspecified role prey prey arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress physical association physical association false false false Fig. 2 GPA1-Cub protein interacts with NubG-GCR1 but not with GCR1-NubG . GCR1-Cub, however, does not show interaction with GPA1-Nub constructs, suggesting that a free GCR1 C-terminus is required for interaction. gcr1-gpa1-3 Interaction detected by Co-IP 143214 N/A 143217 unspecified role unspecified role bait bait arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress N/A 143218 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress physical association physical association false false false Fig. 2 gpa1-gcr1-1 Interaction detected by coimunnoprecipitation 143215 N/A 143217 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region GAL4 activation domain (GAD) tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 143218 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 false in vitro In vitro direct interaction direct interaction false false false Fig. 1 GPA1-GAL4 activation domain fusion protein was used to pull down GCR1 by using anti-GAL4 activation domain gpa1-gcr1-2 Interaction detected by Co-IP 143216 N/A 143218 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress N/A 143217 unspecified role unspecified role prey prey arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress physical association physical association false false false Fig. 2