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 CBP20 targets the cap-binding complex to the nucleus and is stabilized by CBP80. The Arabidopsis CBP20 targets the cap-binding complex to the nucleus and is stabilized by CBP80. Plant J. (0960-7412) 2009 imex curation Kierzkowski D., Kmieciak M., Piontek P., Wojtaszek P., Szweykowska-Kulinska Z., Jarmolowski A. A. thaliana protoplasts were prepared from cell-suspension culture (T87) or from leaves. artjarmo@amu.edu.pl 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 Kierzkowski D., Kmieciak M., Piontek P., Wojtaszek P., Szweykowska-Kulinska Z., Jarmolowski A. A. thaliana protoplasts were prepared from cell-suspension culture (T87) or from leaves. artjarmo@amu.edu.pl Accepted 2009-SEP-11 by ORCHARD. Plant J. (0960-7412) 2009 orchard Only protein-protein interactions imex curation The Arabidopsis CBP20 targets the cap-binding complex to the nucleus and is stabilized by CBP80. The Arabidopsis CBP20 targets the cap-binding complex to the nucleus and is stabilized by CBP80. Plant J. (0960-7412) 2009 imex curation Kierzkowski D., Kmieciak M., Piontek P., Wojtaszek P., Szweykowska-Kulinska Z., Jarmolowski A. A. thaliana protoplasts were prepared from cell-suspension culture (T87) or from leaves. artjarmo@amu.edu.pl orchard Only protein-protein interactions arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Kierzkowski D., Kmieciak M., Piontek P., Wojtaszek P., Szweykowska-Kulinska Z., Jarmolowski A. A. thaliana protoplasts were prepared from cell-suspension culture (T87) or from leaves. artjarmo@amu.edu.pl Accepted 2009-SEP-11 by ORCHARD. Plant J. (0960-7412) 2009 orchard Only protein-protein interactions imex curation The Arabidopsis CBP20 targets the cap-binding complex to the nucleus and is stabilized by CBP80. The Arabidopsis CBP20 targets the cap-binding complex to the nucleus and is stabilized by CBP80. Plant J. (0960-7412) 2009 imex curation Kierzkowski D., Kmieciak M., Piontek P., Wojtaszek P., Szweykowska-Kulinska Z., Jarmolowski A. A. thaliana protoplasts were prepared from cell-suspension culture (T87) or from leaves. artjarmo@amu.edu.pl orchard Only protein-protein interactions arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Kierzkowski D., Kmieciak M., Piontek P., Wojtaszek P., Szweykowska-Kulinska Z., Jarmolowski A. A. thaliana protoplasts were prepared from cell-suspension culture (T87) or from leaves. artjarmo@amu.edu.pl Accepted 2009-SEP-11 by ORCHARD. Plant J. (0960-7412) 2009 orchard Only protein-protein interactions imex curation ncbp2_arath Nuclear cap-binding protein subunit 2 CBP20 At5g44200 20 kDa nuclear cap-binding protein NCBP 20 kDa subunit MLN1.12 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MASLFKEQAKLSAYRDRRFSGTQEEFDEALRASTTVYIGNVSFYTTEEQLYELFSRAGEIKKIIMGLDKNTKTPCGFCFVLFYSREDTEDAVKYISGTILDDRPIRVDFDWGFQEGRQWGRGRSGGQVRDEYRTDYDPARGGYGKLVQKELEAQRQLVDYGTGSLGAYPQAAPTNYGNGRRGGGNYGQGGQNRHGRGDYHRKRQRDDDRYGRDNSRRNTDHESRRDTDSDMRPEKNPRFRESGDSDDDGEDDRKRRS DD8B12A0639E1D58 ncbp1_arath Nuclear cap-binding protein subunit 1 ABH1 CBP80 ENS Abscisic acid-hypersensitive protein 1 Protein ENSALADA T10F5.8 80 kDa nuclear cap-binding protein At2g13540 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSNWKTLLLRIGEKGPEYGTSSDYKDHIETCFGVIRREIERSGDQVLPFLLQCAEQLPHKIPLYGTLIGLLNLENEDFVQKLVESVHANFQVALDSGNCNSIRILLRFMTSLLCSKVIQPASLIVVFETLLSSAATTVDEEKGNPSWQPQADFYVICILSSLPWGGSELAEQVPDEIERVLVGIQAYLSIRKNSSTSGLNFFHNGEFESSLAEKDFVEDLLDRIQSLASNGWKLESVPRPHLSFEAQLVAGKFHELRPIKCMEQPSPPSDHSRAYSGKQKHDALTRYPQRIRRLNIFPANKMEDVQPIDRFVVEEYLLDVLFYLNGCRKECASYMANLPVTFRYEYLMAETLFSQILLLPQPPFKTLYYTLVIMDLCKALPGAFPAVVAGAVRALFEKISDLDMESRTRLILWFSHHLSNFQFIWPWEEWAFVLDLPKWAPKRVFVQEILQREVRLSYWDKIKQSIENATALEELLPPKAGPNFMYSLEEGKEKTEEQQLSAELSRKVKEKQTARDMIVWIEETIYPVHGFEVTLTIVVQTLLDIGSKSFTHLVTVLERYGQVFSKLCPDNDKQVMLLSQVSTYWKNNVQMTAVAIDRMMGYRLVSNQAIVRWVFSPENVDQFHVSDQPWEILGNALNKTYNRISDLRKDISNITKNVLVAEKASANARVELEAAESKLSLVEGEPVLGENPAKMKRLKSTVEKTGEAELSLRESLEAKEALLNRALSETEVLLLLLFQSFLGVLKERLPDPTKVRSVQDLKSIGAEDDKPSAMDVDSENGNPKKSCEVGEREQWCLSTLGYLTAFTRQYASEIWPHMEKLESEVFSGEDVHPLFLQAISSALQFPLH 1E583391F4BAE1A8 cbp20-cbp80 1 5 unspecified role unspecified role prey prey 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 physical association physical association Figure 3 AtCBP20 and AtCBP80 interaction study using the yeast two-hybrid system. cbp20-cbp80-1 2 5 unspecified role unspecified role neutral component neutral component n-terminal Enhanced yellow fluorescent protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 4 unspecified role unspecified role neutral component neutral component n-terminal Enhanced cyan fluorescent protein cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range colocalization colocalization Figure 2 n the next step, protoplasts were co-transfected with fluorescently tagged AtCBP20 and AtCBP80. Both proteins were detected exclusively in the nucleus in the double-transfected cells cbp20-cbp80-2 3 5 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept n-terminal Enhanced yellow fluorescent protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 4 unspecified role unspecified role fluorescence donor fluorescence donor n-terminal Enhanced cyan fluorescent protein cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range direct interaction direct interaction Figure 4 Thus, the results of the FRET analysis confirmed the observations from the yeast two-hybrid analysis that the first 144 aa of AtCBP20 is responsible for the interaction with AtCBP80.