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/ The Arabidopsis cupin domain protein AtPirin1 interacts with the G protein alpha-subunit GPA1 and regulates seed germination and early seedling development. The Arabidopsis cupin domain protein AtPirin1 interacts with the G protein alpha-subunit GPA1 and regulates seed germination and early seedling development. Plant Cell (1040-4651) 2003 Lapik YR., Kaufman LS. The identities of ETR1 and CTR1 were obtained from references given in the text. lkaufman@uic.edu 17-April-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 predetermined predetermined participant predetermined Lapik YR., Kaufman LS. The identities of ETR1 and CTR1 were obtained from references given in the text. lkaufman@uic.edu Accepted 2008-MAR-5 by ORCHARD. 17-April-2008: Contacted by JYOTI. Plant Cell (1040-4651) 2003 orchard The Arabidopsis cupin domain protein AtPirin1 interacts with the G protein alpha-subunit GPA1 and regulates seed germination and early seedling development. The Arabidopsis cupin domain protein AtPirin1 interacts with the G protein alpha-subunit GPA1 and regulates seed germination and early seedling development. Plant Cell (1040-4651) 2003 Lapik YR., Kaufman LS. The identities of ETR1 and CTR1 were obtained from references given in the text. lkaufman@uic.edu 17-April-2008: Contacted by JYOTI. orchard in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Lapik YR., Kaufman LS. The identities of ETR1 and CTR1 were obtained from references given in the text. lkaufman@uic.edu Accepted 2008-MAR-5 by ORCHARD. 17-April-2008: Contacted by JYOTI. Plant Cell (1040-4651) 2003 orchard The Arabidopsis cupin domain protein AtPirin1 interacts with the G protein alpha-subunit GPA1 and regulates seed germination and early seedling development. The Arabidopsis cupin domain protein AtPirin1 interacts with the G protein alpha-subunit GPA1 and regulates seed germination and early seedling development. Plant Cell (1040-4651) 2003 Lapik YR., Kaufman LS. The identities of ETR1 and CTR1 were obtained from references given in the text. lkaufman@uic.edu 17-April-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 Lapik YR., Kaufman LS. Details of the cDNA library were obtained from the Arabidopsis Biological Resource Center. The identities of ETR1 and CTR1 were obtained from references given in the text. lkaufman@uic.edu Accepted 2008-MAR-5 by ORCHARD. 17-April-2008: Contacted by JYOTI. cDNA library of 3 day-old etiolated arabidopsis seedlings, (ecotype Columbia). Plant Cell (1040-4651) 2003 orchard ctr1_arath Serine/threonine-protein kinase CTR1 Protein CONSTITUTIVE TRIPLE RESPONSE1 F17C15_150 CTR1 At5g03730 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEMPGRRSNYTLLSQFSDDQVSVSVTGAPPPHYDSLSSENRSNHNSGNTGKAKAERGGFDWDPSGGGGGDHRLNNQPNRVGNNMYASSLGLQRQSSGSSFGESSLSGDYYMPTLSAAANEIESVGFPQDDGFRLGFGGGGGDLRIQMAADSAGGSSSGKSWAQQTEESYQLQLALALRLSSEATCADDPNFLDPVPDESALRTSPSSAETVSHRFWVNGCLSYYDKVPDGFYMMNGLDPYIWTLCIDLHESGRIPSIESLRAVDSGVDSSLEAIIVDRRSDPAFKELHNRVHDISCSCITTKEVVDQLAKLICNRMGGPVIMGEDELVPMWKECIDGLKEIFKVVVPIGSLSVGLCRHRALLFKVLADIIDLPCRIAKGCKYCNRDDAASCLVRFGLDREYLVDLVGKPGHLWEPDSLLNGPSSISISSPLRFPRPKPVEPAVDFRLLAKQYFSDSQSLNLVFDPASDDMGFSMFHRQYDNPGGENDALAENGGGSLPPSANMPPQNMMRASNQIEAAPMNAPPISQPVPNRANRELGLDGDDMDIPWCDLNIKEKIGAGSFGTVHRAEWHGSDVAVKILMEQDFHAERVNEFLREVAIMKRLRHPNIVLFMGAVTQPPNLSIVTEYLSRGSLYRLLHKSGAREQLDERRRLSMAYDVAKGMNYLHNRNPPIVHRDLKSPNLLVDKKYTVKVCDFGLSRLKASTFLSSKSAAGTPEWMAPEVLRDEPSNEKSDVYSFGVILWELATLQQPWGNLNPAQVVAAVGFKCKRLEIPRNLNPQVAAIIEGCWTNEPWKRPSFATIMDLLRPLIKSAVPPPNRSDL 2922D3DCD0CC15BC 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 DIP protein P18064 original sequence version: 80 46BF8650E067F968 prn1_arath Pirin-1 PRN1 F25L23_80 At3g59220 AtPirin1 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MTYENNSVPRIVIKKVLAKLEKEGEGAVVRNGITKIDQKLLDPFVLLVEFSFSLSAGFPDHPHRGFESVTYMLQGGIIHKDPKGHKGTIQAGDVQWMTAGRGIIHSEFPEEEVNNGLQLWINLPSTEKMTEPKYKELSSLDIPRAEENGVEVKVIAGDSMGIKSPVYTRTPTMFLDFTLKPGSQTHQTVPESWTAFAYIIEGDEGVFGSLNSSAISAHHVVVFGPGDLVSVWNKSTSRSLRFLLIAGEPIGEPVVQCGPFVMNSQAEIDMAFDDYQNAKNGFEMAKC F44ACE2F98208B1E etr1_arath Ethylene receptor 1 ETR1 T27F4.9 At1g66340 Protein ETHYLENE RESPONSE 1 Protein ETR1 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEVCNCIEPQWPADELLMKYQYISDFFIAIAYFSIPLELIYFVKKSAVFPYRWVLVQFGAFIVLCGATHLINLWTFTTHSRTVALVMTTAKVLTAVVSCATALMLVHIIPDLLSVKTRELFLKNKAAELDREMGLIRTQEETGRHVRMLTHEIRSTLDRHTILKTTLVELGRTLALEECALWMPTRTGLELQLSYTLRHQHPVEYTVPIQLPVINQVFGTSRAVKISPNSPVARLRPVSGKYMLGEVVAVRVPLLHLSNFQINDWPELSTKRYALMVLMLPSDSARQWHVHELELVEVVADQVAVALSHAAILEESMRARDLLMEQNVALDLARREAETAIRARNDFLAVMNHEMRTPMHAIIALSSLLQETELTPEQRLMVETILKSSNLLATLMNDVLDLSRLEDGSLQLELGTFNLHTLFREVLNLIKPIAVVKKLPITLNLAPDLPEFVVGDEKRLMQIILNIVGNAVKFSKQGSISVTALVTKSDTRAADFFVVPTGSHFYLRVKVKDSGAGINPQDIPKIFTKFAQTQSLATRSSGGSGLGLAISKRFVNLMEGNIWIESDGLGKGCTAIFDVKLGISERSNESKQSGIPKVPAIPRHSNFTGLKVLVMDENGVSRMVTKGLLVHLGCEVTTVSSNEECLRVVSHEHKVVFMDVCMPGVENYQIALRIHEKFTKQRHQRPLLVALSGNTDKSTKEKCMSFGLDGVLLKPVSLDNIRDVLSDLLEPRVLYEGM D6ED3C4BBA87E96E gpa1-prn1 1 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait physical association physical association Figure 1A Reconstitution of the yeast two-hybrid interaction between AtPirin1 and GPA1. etr1-ctr1 1 4 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Figure 1A The interaction between the Arabidopsis proteins ETR1 and CTR1 (Clark et al., 1998) served as a positive control for the assay (Figure 1A). gpa1-prn1-1 2 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 in vitro In vitro 5 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) physical association physical association Figure 1C In vitro protein interaction between AtPirin1 and GPA1. gpa1-prn1-2 3 6 unspecified role unspecified role prey prey c-terminal binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role bait bait physical association physical association Therefore, we named this GPA1-interacting protein AtPirin1