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/ Identification and characterization of proteins that selectively interact with isoforms of the mRNA binding protein AUF1 (hnRNP D). Identification and characterization of proteins that selectively interact with isoforms of the mRNA binding protein AUF1 (hnRNP D). Biol. Chem. (1431-6730) 2003 Moraes KC., Quaresma AJ., Maehnss K., Kobarg J. Synapse - Interactions of proteins with an established role in the presynapse. 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 Moraes KC., Quaresma AJ., Maehnss K., Kobarg J. Synapse - Interactions of proteins with an established role in the presynapse. Accepted 2006-OCT-12 by JYOTI. Biol. Chem. (1431-6730) 2003 Identification and characterization of proteins that selectively interact with isoforms of the mRNA binding protein AUF1 (hnRNP D). Identification and characterization of proteins that selectively interact with isoforms of the mRNA binding protein AUF1 (hnRNP D). Biol. Chem. (1431-6730) 2003 Moraes KC., Quaresma AJ., Maehnss K., Kobarg J. Synapse - Interactions of proteins with an established role in the presynapse. 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 Moraes KC., Quaresma AJ., Maehnss K., Kobarg J. Synapse - Interactions of proteins with an established role in the presynapse. Accepted 2006-OCT-12 by JYOTI. a human fetal brain cDNA library was used. Biol. Chem. (1431-6730) 2003 Identification and characterization of proteins that selectively interact with isoforms of the mRNA binding protein AUF1 (hnRNP D). Identification and characterization of proteins that selectively interact with isoforms of the mRNA binding protein AUF1 (hnRNP D). Biol. Chem. (1431-6730) 2003 Moraes KC., Quaresma AJ., Maehnss K., Kobarg J. Synapse - Interactions of proteins with an established role in the presynapse. 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 Moraes KC., Quaresma AJ., Maehnss K., Kobarg J. Synapse - Interactions of proteins with an established role in the presynapse. Accepted 2006-OCT-12 by JYOTI. Biol. Chem. (1431-6730) 2003 Identification and characterization of proteins that selectively interact with isoforms of the mRNA binding protein AUF1 (hnRNP D). Identification and characterization of proteins that selectively interact with isoforms of the mRNA binding protein AUF1 (hnRNP D). Biol. Chem. (1431-6730) 2003 Moraes KC., Quaresma AJ., Maehnss K., Kobarg J. Synapse - Interactions of proteins with an established role in the presynapse. in vitro In vitro pull down pull down predetermined predetermined participant predetermined Moraes KC., Quaresma AJ., Maehnss K., Kobarg J. Synapse - Interactions of proteins with an established role in the presynapse. Accepted 2006-OCT-12 by JYOTI. Biol. Chem. (1431-6730) 2003 Identification and characterization of proteins that selectively interact with isoforms of the mRNA binding protein AUF1 (hnRNP D). Identification and characterization of proteins that selectively interact with isoforms of the mRNA binding protein AUF1 (hnRNP D). Biol. Chem. (1431-6730) 2003 Moraes KC., Quaresma AJ., Maehnss K., Kobarg J. Synapse - Interactions of proteins with an established role in the presynapse. in vitro In vitro emsa supershift electrophoretic mobility supershift assay emsa supershift predetermined predetermined participant predetermined Moraes KC., Quaresma AJ., Maehnss K., Kobarg J. Synapse - Interactions of proteins with an established role in the presynapse. Accepted 2006-OCT-12 by JYOTI. Biol. Chem. (1431-6730) 2003 ubc9_human SUMO-conjugating enzyme UBC9 UBE2I p18 SUMO-protein ligase UBC9 UBCE9 Ubiquitin carrier protein 9 Ubiquitin carrier protein I Ubiquitin-conjugating enzyme E2 I Ubiquitin-protein ligase I RING-type E3 SUMO transferase UBC9 protein protein human Homo sapiens Human MSGIALSRLAQERKAWRKDHPFGFVAVPTKNPDGTMNLMNWECAIPGKKGTPWEGGLFKLRMLFKDDYPSSPPKCKFEPPLFHPNVYPSGTVCLSILEEDKDWRPAITIKQILLGIQELLNEPNIQDPAQAEAYTIYCQNRVEYEKRVRAQAKKFAPS DIP protein P63279 original sequence version: 33 DIP protein P63279 original sequence version: 57 DIP protein P63279 original sequence version: 80 DIP protein P63279 original sequence version: 137 DIP protein P63279 original sequence version: 99 DIP protein P63279 original sequence version: 140 DIP protein P63279 original sequence version: 120 DIP protein P63279 original sequence version: 117 DIP protein P63279 original sequence version: 93 E2C826E9C8D0683D q14103-3 Heterogeneous nuclear ribonucleoprotein D0 p40 Dx7 HNRNPD AUF1 HNRPD AU-rich element RNA-binding protein 1 protein protein human Homo sapiens Human MSEEQFGGDGAAAAATAAVGGSAGEQEGAMVAATQGAAAAAGSGAGTGGGTASGGTEGGSAESEGAKIDASKNEEDEGHSNSSPRHSEAATAQREEWKMFIGGLSWDTTKKDLKDYFSKFGEVVDCTLKLDPITGRSRGFGFVLFKESESVDKVMDQKEHKLNGKVIDPKRAKAMKTKEPVKKIFVGGLSPDTPEEKIREYFGGFGEVESIELPMDNKTNKRRGFCFITFKEEEPVKKIMEKKYHNVGLSKCEIKVAMSKEQYQQQQQWGSRGGFAGRARGRGGDQQSGYGKVSRRGGHQNSYKPY Contains a omega-N-methylarginine at position 282. Contains a N6-acetyllysine at position 292. ABCDD6ACF812F647 hnrpq_human Heterogeneous nuclear ribonucleoprotein Q Synaptotagmin-binding, cytoplasmic RNA-interacting protein Glycine- and tyrosine-rich RNA-binding protein NS1-associated protein 1 SYNCRIP HNRPQ NSAP1 protein protein human Homo sapiens Human MATEHVNGNGTEEPMDTTSAVIHSENFQTLLDAGLPQKVAEKLDEIYVAGLVAHSDLDERAIEALKEFNEDGALAVLQQFKDSDLSHVQNKSAFLCGVMKTYRQREKQGTKVADSSKGPDEAKIKALLERTGYTLDVTTGQRKYGGPPPDSVYSGQQPSVGTEIFVGKIPRDLFEDELVPLFEKAGPIWDLRLMMDPLTGLNRGYAFVTFCTKEAAQEAVKLYNNHEIRSGKHIGVCISVANNRLFVGSIPKSKTKEQILEEFSKVTEGLTDVILYHQPDDKKKNRGFCFLEYEDHKTAAQARRRLMSGKVKVWGNVGTVEWADPIEDPDPEVMAKVKVLFVRNLANTVTEEILEKAFSQFGKLERVKKLKDYAFIHFDERDGAVKAMEEMNGKDLEGENIEIVFAKPPDQKRKERKAQRQAAKNQMYDDYYYYGPPHMPPPTRGRGRGGRGGYGYPPDYYGYEDYYDYYGYDYHNYRGGYEDPYYGYEDFQVGARGRGGRGARGAAPSRGRGAAPPRGRAGYSQRGGPGSARGVRGARGGAQQQRGRGVRGARGGRGGNVGGKRKADGYNQPDSKRRQTNNQNWGSQPIAQQPLQGGDHSGNYGYKSENQEFYQDTFGQQWK DIP protein O60506 original sequence version: 144 DIP protein O60506 original sequence version: 124 0669FA604E8FBBDF o60506-1 Heterogeneous nuclear ribonucleoprotein Q hnRNP Q3 SYNCRIP HNRPQ NSAP1 Glycine- and tyrosine-rich RNA-binding protein NS1-associated protein 1 Synaptotagmin-binding, cytoplasmic RNA-interacting protein protein protein human Homo sapiens Human MATEHVNGNGTEEPMDTTSAVIHSENFQTLLDAGLPQKVAEKLDEIYVAGLVAHSDLDERAIEALKEFNEDGALAVLQQFKDSDLSHVQNKSAFLCGVMKTYRQREKQGTKVADSSKGPDEAKIKALLERTGYTLDVTTGQRKYGGPPPDSVYSGQQPSVGTEIFVGKIPRDLFEDELVPLFEKAGPIWDLRLMMDPLTGLNRGYAFVTFCTKEAAQEAVKLYNNHEIRSGKHIGVCISVANNRLFVGSIPKSKTKEQILEEFSKVTEGLTDVILYHQPDDKKKNRGFCFLEYEDHKTAAQARRRLMSGKVKVWGNVGTVEWADPIEDPDPEVMAKVKVLFVRNLANTVTEEILEKAFSQFGKLERVKKLKDYAFIHFDERDGAVKAMEEMNGKDLEGENIEIVFAKPPDQKRKERKAQRQAAKNQMYDDYYYYGPPHMPPPTRGRGRGGRGGYGYPPDYYGYEDYYDYYGYDYHNYRGGYEDPYYGYEDFQVGARGRGGRGARGAAPSRGRGAAPPRGRAGYSQRGGPGSARGVRGARGGAQQQRGRGVRGARGGRGGNVGGKRKADGYNQPDSKRRQTNNQNWGSQPIAQQPLQGGDHSGNYGYKSENQEFYQDTFGQQWK 0669FA604E8FBBDF if2b2_human Insulin-like growth factor 2 mRNA-binding protein 2 IGF-II mRNA-binding protein 2 Hepatocellular carcinoma autoantigen p62 VICKZ family member 2 IGF2BP2 IMP2 VICKZ2 protein protein human Homo sapiens Human MMNKLYIGNLSPAVTADDLRQLFGDRKLPLAGQVLLKSGYAFVDYPDQNWAIRAIETLSGKVELHGKIMEVDYSVSKKLRSRKIQIRNIPPHLQWEVLDGLLAQYGTVENVEQVNTDTETAVVNVTYATREEAKIAMEKLSGHQFENYSFKISYIPDEEVSSPSPPQRAQRGDHSSREQGHAPGGTSQARQIDFPLRILVPTQFVGAIIGKEGLTIKNITKQTQSRVDIHRKENSGAAEKPVTIHATPEGTSEACRMILEIMQKEADETKLAEEIPLKILAHNGLVGRLIGKEGRNLKKIEHETGTKITISSLQDLSIYNPERTITVKGTVEACASAEIEIMKKLREAFENDMLAVNQQANLIPGLNLSALGIFSTGLSVLSPPAGPRGAPPAAPYHPFTTHSGYFSSLYPHHQFGPFPHHHSYPEQEIVNLFIPTQAVGAIIGKKGAHIKQLARFAGASIKIAPAEGPDVSERMVIITGPPEAQFKAQGRIFGKLKEENFFNPKEEVKLEAHIRVPSSTAGRVIGKGGKTVNELQNLTSAEVIVPRDQTPDENEEVIVRIIGHFFASQTAQRKIREIVQQVKQQEQKYPQGVASQRSK DIP protein Q9Y6M1 original sequence version: 149 EA656F8733BBB39A 38mer rna ribonucleic acid RNA rna human Homo sapiens Human GUGAUUAUUUAUUAUUUAUUUAUUAUUUAUUUAUUUAG 1136CC117DC5B48C ybox1_human Y-box-binding protein 1 YBX1 YB1 NSEP1 Y-box transcription factor CCAAT-binding transcription factor I subunit A Enhancer factor I subunit A DNA-binding protein B Nuclease-sensitive element-binding protein 1 protein protein human Homo sapiens Human MSSEAETQQPPAAPPAAPALSAADTKPGTTGSGAGSGGPGGLTSAAPAGGDKKVIATKVLGTVKWFNVRNGYGFINRNDTKEDVFVHQTAIKKNNPRKYLRSVGDGETVEFDVVEGEKGAEAANVTGPGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNYQNSESGEKNEGSESAPEGQAQQRRPYRRRRFPPYYMRRPYGRRPQYSNPPVQGEVMEGADNQGAGEQGRPVRQNMYRGYRPRFRRGPPRQRQPREDGNEEDKENQGDETQGQQPPQRRYRRNFNYRRRRPENPKPQDGKETKAADPPAENSSAPEAEQGGAE DIP protein P67809 original sequence version: 39 DF0114BF974AEDB8 q14103-4 Heterogeneous nuclear ribonucleoprotein D0 p37 HNRNPD AUF1 HNRPD AU-rich element RNA-binding protein 1 protein protein human Homo sapiens Human MSEEQFGGDGAAAAATAAVGGSAGEQEGAMVAATQGAAAAAGSGAGTGGGTASGGTEGGSAESEGAKIDASKNEEDEGKMFIGGLSWDTTKKDLKDYFSKFGEVVDCTLKLDPITGRSRGFGFVLFKESESVDKVMDQKEHKLNGKVIDPKRAKAMKTKEPVKKIFVGGLSPDTPEEKIREYFGGFGEVESIELPMDNKTNKRRGFCFITFKEEEPVKKIMEKKYHNVGLSKCEIKVAMSKEQYQQQQQWGSRGGFAGRARGRGGDQQSGYGKVSRRGGHQNSYKPY Contains a omega-N-methylarginine at position 263. Contains a N6-acetyllysine at position 273. 98DF6E78EAF3BBC1 hnrpd_human Heterogeneous nuclear ribonucleoprotein D0 HNRNPD AUF1 HNRPD AU-rich element RNA-binding protein 1 protein protein human Homo sapiens Human MSEEQFGGDGAAAAATAAVGGSAGEQEGAMVAATQGAAAAAGSGAGTGGGTASGGTEGGSAESEGAKIDASKNEEDEGHSNSSPRHSEAATAQREEWKMFIGGLSWDTTKKDLKDYFSKFGEVVDCTLKLDPITGRSRGFGFVLFKESESVDKVMDQKEHKLNGKVIDPKRAKAMKTKEPVKKIFVGGLSPDTPEEKIREYFGGFGEVESIELPMDNKTNKRRGFCFITFKEEEPVKKIMEKKYHNVGLSKCEIKVAMSKEQYQQQQQWGSRGGFAGRARGRGGGPSQNWNQGYSNYWNQGYGNYGYNSQGYGGYGGYDYTGYNNYYGYGDYSNQQSGYGKVSRRGGHQNSYKPY DIP protein Q14103 original sequence version: 199 DIP protein Q14103 original sequence version: 164 D0B6EA177BEF789E hnrpd-syncrip 3 13 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. Co-transformation experiments of the yeast strain L40 with pBTM116-AUF1- p37, -p40, -p42 and -p45 and pACT2-NSEP-1, pACT2-NSAP-1 and pACT2-IMP-2 (Figure 1C) demonstrated that the three RNA binding proteins NSEP-1, NSAP-1 and IMP-2 all interact only with AUF1p37 and p40 but not with p42 or p45. In contrast, protein UBCE2I interacted with all four isoforms. 1C NSAP-1 interacted only with AUF1p37 and p40 but not with p42 or p45. hnrpd-ybx1 3 13 unspecified role unspecified role bait bait 12 unspecified role unspecified role prey prey physical association physical association AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. Co-transformation experiments of the yeast strain L40 with pBTM116-AUF1- p37, -p40, -p42 and -p45 and pACT2-NSEP-1, pACT2-NSAP-1 and pACT2-IMP-2 (Figure 1C) demonstrated that the three RNA binding proteins NSEP-1, NSAP-1 and IMP-2 all interact only with AUF1p37 and p40 but not with p42 or p45. In contrast, protein UBCE2I interacted with all four isoforms. 1C NSEP-1 interacted only with AUF1p37 and p40 but not with p42 or p45. hnrpd-ube2i 3 6 unspecified role unspecified role prey prey 14 unspecified role unspecified role bait bait physical association physical association AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. Co-transformation experiments of the yeast strain L40 with pBTM116-AUF1- p37, -p40, -p42 and -p45 and pACT2-NSEP-1, pACT2-NSAP-1 and pACT2-IMP-2 (Figure 1C) demonstrated that the three RNA binding proteins NSEP-1, NSAP-1 and IMP-2 all interact only with AUF1p37 and p40 but not with p42 or p45. In contrast, protein UBCE2I interacted with all four isoforms. 1C protein UBCE2I interacted with all four isoforms. hnrpd-igf2bp2 1 13 unspecified role unspecified role bait bait ecobb Escherichia coli (strain B / BL21) 10 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1024556]The shifted ranges (248-510) couldn't be applied as the new full feature sequence is different from the previous one. [EBI-1024556]248-467 [EBI-1024556]Q9Y6M1,1 [EBI-1024557]The shifted ranges (331-599) couldn't be applied as the new full feature sequence is different from the previous one. [EBI-1024557]331-556 [EBI-1024557]Q9Y6M1,1 physical association physical association AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. the two constructs containing the KH domains 2 and 3 or 3 and 4 were able to interact with AUF1. 3 hnrpd-ybx1-1 4 13 unspecified role unspecified role prey prey region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) 12 unspecified role unspecified role bait bait n terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. Next we performed in vitro pull-down assays to verify the observed interactions between AUF1 and the proteins NSAP-1, NSEP-2, IMP-2 and UBCE2I.All four proteins interacted in vitro with AUF1p37. 2 NSEP-1 interact with AUF1p37 invitro. hnrpd-igf2bp2-1 4 13 unspecified role unspecified role prey prey region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) 10 unspecified role unspecified role bait bait n terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. Next we performed in vitro pull-down assays to verify the observed interactions between AUF1 and the proteins NSAP-1, NSEP-2, IMP-2 and UBCE2I.All four proteins interacted in vitro with AUF1p37. 2 IMP-2 interact with AUF1p37 invitro. hnrpd-ube2i-1 4 6 unspecified role unspecified role bait bait n terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecobb Escherichia coli (strain B / BL21) 13 unspecified role unspecified role prey prey region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. Next we performed in vitro pull-down assays to verify the observed interactions between AUF1 and the proteins NSAP-1, NSEP-2, IMP-2 and UBCE2I.All four proteins interacted in vitro with AUF1p37. 2 UBCE2I interact with AUF1p37 invitro. hnrpd-syncrip-1 4 9 unspecified role unspecified role bait bait n terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecobb Escherichia coli (strain B / BL21) 13 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. Next we performed in vitro pull-down assays to verify the observed interactions between AUF1 and the proteins NSAP-1, NSEP-2, IMP-2 and UBCE2I.All four proteins interacted in vitro with AUF1p37. 2 NSAP-1 interact with AUF1p37 invitro. hnrpd-syncrip-2 1 8 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain 13 unspecified role unspecified role bait bait physical association physical association AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. Our results show that the construct that encompasses the RRM domains 3 and 4, as well as that which contains domains 2 and 3, both interact with AUF1p37, whereas a construct containing domains 1 and 2 did not interact. 3 These results show that at least two domains of NSAP1 are required for the interaction with AUF1p37 and that domain 3 alone is not sufficient for the interaction. hnrpd-ybx1-3 1 12 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain 13 unspecified role unspecified role bait bait physical association physical association AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. These results suggest that more than two of the B/A islands are required for the interaction with AUF1. Two constructs that encompass three B/A islands (1-2-3 and 2-3-4) were both able to interact with AUF1p37. 3 hnrpd-igf2bp2-2 3 10 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait physical association physical association AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. Co-transformation experiments of the yeast strain L40 with pBTM116-AUF1- p37, -p40, -p42 and -p45 and pACT2-NSEP-1, pACT2-NSAP-1 and pACT2-IMP-2 (Figure 1C) demonstrated that the three RNA binding proteins NSEP-1, NSAP-1 and IMP-2 all interact only with AUF1p37 and p40 but not with p42 or p45. In contrast, protein UBCE2I interacted with all four isoforms. 1c IMP-2 interacted only with AUF1p37 and p40 but not with p42 or p45. igf2bp2-38mer 5 10 unspecified role unspecified role neutral component neutral component n terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecobb Escherichia coli (strain B / BL21) 11 unspecified role unspecified role neutral component neutral component 5 primer radiolabelled radiolabel radiolabeled radiolabelled certain certain sequence position certain certain certain sequence position certain in vitro In vitro physical association physical association Binding of GST-IMP-2 to the AU-rich RNA oligonucleotide. 4D (B-E) Binding of the interacting proteins both in the absence (nil) and presence of 52 nM 6xHis-AUF1p37. syncrip-38mer-1 5 8 unspecified role unspecified role neutral component neutral component n terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 11 unspecified role unspecified role neutral component neutral component 5 primer radiolabelled radiolabel radiolabeled radiolabelled certain certain sequence position certain certain certain sequence position certain physical association physical association (B)Binding of 6xHis-NSAP-1 to the AU-rich RNA oligonucleotide. 4B (B-E) Binding of the interacting proteins both in the absence (nil) and presence of 52 nM 6xHis-AUF1p37. ybx1-38mer-1 5 12 unspecified role unspecified role neutral component neutral component n terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecobb Escherichia coli (strain B / BL21) 11 unspecified role unspecified role neutral component neutral component 5 primer radiolabelled radiolabel radiolabeled radiolabelled certain certain sequence position certain certain certain sequence position certain in vitro In vitro physical association physical association Binding of GST-NSEP-1 to the AU-rich RNA oligonucleotide. 4C (B-E) Binding of the interacting proteins both in the absence (nil) and presence of 52 nM 6xHis-AUF1p37. ube2i-38mer 5 6 unspecified role unspecified role neutral component neutral component n terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecobb Escherichia coli (strain B / BL21) 11 unspecified role unspecified role neutral component neutral component 5 primer radiolabelled radiolabel radiolabeled radiolabelled certain certain sequence position certain certain certain sequence position certain in vitro In vitro physical association physical association Binding of GST-UBCE2I to the AU-rich RNA oligonucleotide. 4E (B-E) Binding of the interacting proteins both in the absence (nil) and presence of 52 nM 6xHis-AUF1p37. hnrpd-38mer 5 11 unspecified role unspecified role neutral component neutral component 5 primer radiolabelled radiolabel radiolabeled radiolabelled certain certain sequence position certain certain certain sequence position certain in vitro In vitro 13 unspecified role unspecified role neutral component neutral component n terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecobb Escherichia coli (strain B / BL21) physical association physical association AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1. p42 IsoformId:Q14103-2. p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.AUF1 p37 isoform was used in the experiment. Binding of 6His-AUF1 to the AU-rich RNA oligonucleotide. 4A hnrpd-syncrip-3 3 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. Co-transformation experiments of the yeast strain L40 with pBTM116-AUF1- p37, -p40, -p42 and -p45 and pACT2-NSEP-1, pACT2-NSAP-1 and pACT2-IMP-2 (Figure 1C) demonstrated that the three RNA binding proteins NSEP-1, NSAP-1 and IMP-2 all interact only with AUF1p37 and p40 but not with p42 or p45. In contrast, protein UBCE2I interacted with all four isoforms. 1C NSAP-1 interacted only with AUF1p37 and p40 but not with p42 or p45. hnrpd-ybx1-4 3 12 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. Co-transformation experiments of the yeast strain L40 with pBTM116-AUF1- p37, -p40, -p42 and -p45 and pACT2-NSEP-1, pACT2-NSAP-1 and pACT2-IMP-2 (Figure 1C) demonstrated that the three RNA binding proteins NSEP-1, NSAP-1 and IMP-2 all interact only with AUF1p37 and p40 but not with p42 or p45. In contrast, protein UBCE2I interacted with all four isoforms. 1C NSEP-1 interacted only with AUF1p37 and p40 but not with p42 or p45. hnrpd-igf2bp2-3 3 10 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1. p42 IsoformId:Q14103-2. p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. Co-transformation experiments of the yeast strain L40 with pBTM116-AUF1- p37, -p40, -p42 and -p45 and pACT2-NSEP-1, pACT2-NSAP-1 and pACT2-IMP-2 (Figure 1C) demonstrated that the three RNA binding proteins NSEP-1, NSAP-1 and IMP-2 all interact only with AUF1p37 and p40 but not with p42 or p45. In contrast, protein UBCE2I interacted with all four isoforms. 1c IMP-2 interacted only with AUF1p37 and p40 but not with p42 or p45. hnrpd-hnrpd 3 7 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait physical association physical association AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. Our analysis revealed that AUF1p37 only interacts with the AUF1 isoforms p37 and p40 but not with p42 or p45 (Figure 1B). 1B hnrpd-hnrpd-1 3 13 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait physical association physical association AUF1 exists as four different isoforms (p37, p40, p42 and p45) that are generated by alternative splicing. p45 IsoformId:Q14103-1.p42 IsoformId:Q14103-2.p40 IsoformId:Q14103-3. p37 IsoformId:Q14103-4.four isoforms were used in the experiment. Our analysis revealed that AUF1p37 only interacts with the AUF1 isoforms p37 and p40 but not with p42 or p45 (Figure 1B). 1B hnrpd-syncrip-6 2 13 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey n terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Table 1 hnrpd-ybx1-7 2 7 unspecified role unspecified role bait bait 12 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association table 1 hnrpd-igf2bp2-6 2 10 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait physical association physical association table 1 hnrpd-ube2i-2 2 14 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association table 1