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 fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. Cell (0092-8674) 2008 imex curation Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be 14-Nov-2008: Contacted by JYOTI. davet Only protein-protein interactions 11 in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Author contacted - no response to date Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. Data on 4E-T could not be captured as source of protein could not be identified claudia.bagni@med.kuleuven.be Accepted 2008-NOV-7 by DAVET. 14-Nov-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 11 imex curation The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. Cell (0092-8674) 2008 imex curation Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be 14-Nov-2008: Contacted by JYOTI. davet Only protein-protein interactions 11 mouse-neuron Mus musculus neuronal cell neuron Neuron [Neuronal cell; Neural cell fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be Accepted 2008-NOV-7 by DAVET. 14-Nov-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 11 imex curation The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. Cell (0092-8674) 2008 imex curation Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be 14-Nov-2008: Contacted by JYOTI. davet Only protein-protein interactions 11 mouse-neuron Mus musculus neuronal cell neuron Neuron [Neuronal cell; Neural cell anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be Accepted 2008-NOV-7 by DAVET. 14-Nov-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 11 imex curation The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. Cell (0092-8674) 2008 imex curation Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be 14-Nov-2008: Contacted by JYOTI. davet Only protein-protein interactions 11 mouse-brain Mus musculus brain brain Brain [cerebrum] density sedimentation cosedimentation through density gradient density sedimentation western blot western blot Immuno blot Immuno blot Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be Accepted 2008-NOV-7 by DAVET. Cell (0092-8674) 2008 davet Only protein-protein interactions 11 imex curation The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. Cell (0092-8674) 2008 imex curation Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be 14-Nov-2008: Contacted by JYOTI. davet Only protein-protein interactions 11 in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be Accepted 2008-NOV-7 by DAVET. 14-Nov-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 11 imex curation The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. Cell (0092-8674) 2008 imex curation Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be 14-Nov-2008: Contacted by JYOTI. davet Only protein-protein interactions 11 in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip primer specific pcr primer specific pcr Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be Accepted 2008-NOV-7 by DAVET. 14-Nov-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 11 imex curation The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. Cell (0092-8674) 2008 imex curation Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be 14-Nov-2008: Contacted by JYOTI. davet Only protein-protein interactions 11 mouse-brain Mus musculus brain brain Brain [cerebrum] affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom western blot western blot Immuno blot Immuno blot Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be Accepted 2008-NOV-7 by DAVET. 14-Nov-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 11 imex curation The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. Cell (0092-8674) 2008 imex curation Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be 14-Nov-2008: Contacted by JYOTI. davet Only protein-protein interactions 11 mouse-brain Mus musculus brain brain Brain [cerebrum] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be Accepted 2008-NOV-7 by DAVET. 14-Nov-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 11 imex curation The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. The fragile X syndrome protein represses activity-dependent translation through CYFIP1, a new 4E-BP. Cell (0092-8674) 2008 imex curation Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be 14-Nov-2008: Contacted by JYOTI. davet Only protein-protein interactions 11 mouse-neuron Mus musculus neuronal cell neuron Neuron [Neuronal cell; Neural cell anti bait coip anti bait coimmunoprecipitation anti bait coip primer specific pcr primer specific pcr Napoli I., Mercaldo V., Boyl PP., Eleuteri B., Zalfa F., De Rubeis S., Di Marino D., Mohr E., Massimi M., Falconi M., Witke W., Costa-Mattioli M., Sonenberg N., Achsel T., Bagni C. Total mouse brain was homogenized in lysis buffer and lysates were centrifuged. Supernatants centrifuged through 15%-50% (w/v) sucrose gradients for 110 min at 37.000 and 5%-25% for 180 min at 39,000 rpm. claudia.bagni@med.kuleuven.be Accepted 2008-NOV-7 by DAVET. 14-Nov-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 11 imex curation bc1_mouse_rna BC1 RNA rna ribonucleic acid RNA rna mouse Mus musculus Mouse A small untranslated RNA which is a specific effector of translational control. cyfp1_human Cytoplasmic FMR1-interacting protein 1 Specifically Rac1-associated protein 1 p140sra-1 CYFIP1 KIAA0068 protein protein human Homo sapiens Human MAAQVTLEDALSNVDLLEELPLPDQQPCIEPPPSSLLYQPNFNTNFEDRNAFVTGIARYIEQATVHSSMNEMLEEGQEYAVMLYTWRSCSRAIPQVKCNEQPNRVEIYEKTVEVLEPEVTKLMNFMYFQRNAIERFCGEVRRLCHAERRKDFVSEAYLITLGKFINMFAVLDELKNMKCSVKNDHSAYKRAAQFLRKMADPQSIQESQNLSMFLANHNKITQSLQQQLEVISGYEELLADIVNLCVDYYENRMYLTPSEKHMLLKVMGFGLYLMDGSVSNIYKLDAKKRINLSKIDKYFKQLQVVPLFGDMQIELARYIKTSAHYEENKSRWTCTSSGSSPQYNICEQMIQIREDHMRFISELARYSNSEVVTGSGRQEAQKTDAEYRKLFDLALQGLQLLSQWSAHVMEVYSWKLVHPTDKYSNKDCPDSAEEYERATRYNYTSEEKFALVEVIAMIKGLQVLMGRMESVFNHAIRHTVYAALQDFSQVTLREPLRQAIKKKKNVIQSVLQAIRKTVCDWETGHEPFNDPALRGEKDPKSGFDIKVPRRAVGPSSTQLYMVRTMLESLIADKSGSKKTLRSSLEGPTILDIEKFHRESFFYTHLINFSETLQQCCDLSQLWFREFFLELTMGRRIQFPIEMSMPWILTDHILETKEASMMEYVLYSLDLYNDSAHYALTRFNKQFLYDEIEAEVNLCFDQFVYKLADQIFAYYKVMAGSLLLDKRLRSECKNQGATIHLPPSNRYETLLKQRHVQLLGRSIDLNRLITQRVSAAMYKSLELAIGRFESEDLTSIVELDGLLEINRMTHKLLSRYLTLDGFDAMFREANHNVSAPYGRITLHVFWELNYDFLPNYCYNGSTNRFVRTVLPFSQEFQRDKQPNAQPQYLHGSKALNLAYSSIYGSYRNFVGPPHFQVICRLLGYQGIAVVMEELLKVVKSLLQGTILQYVKTLMEVMPKICRLPRHEYGSPGILEFFHHQLKDIVEYAELKTVCFQNLREVGNAILFCLLIEQSLSLEEVCDLLHAAPFQNILPRVHVKEGERLDAKMKRLESKYAPLHLVPLIERLGTPQQIAIAREGDLLTKERLCCGLSMFEVILTRIRSFLDDPIWRGPLPSNGVMHVDECVEFHRLWSAMQFVYCIPVGTHEFTVEQCFGDGLHWAGCMIIVLLGQQRRFAVLDFCYHLLKVQKHDGKDEIIKNVPLKKMVERIRKFQILNDEIITILDKYLKSGDGEGTPVEHVRCFQPPIHQSLASS DIP protein Q7L576 original sequence version: 55 DIP protein Q7L576-1 original sequence version: 59 DIP protein Q7L576 original sequence version: 59 DIP protein Q7L576 original sequence version: 100 D8F45E13207BEF16 arc_mouse_rna Mouse Arc mRNA mrna messenger rna mRNA mrna mouse Mus musculus Mouse a4_mouse_rna Mouse A4 mRNA rna ribonucleic acid RNA rna mouse Mus musculus Mouse rs6_mouse 40S ribosomal protein S6 Rps6 Phosphoprotein NP33 protein protein mouse Mus musculus Mouse MKLNISFPATGCQKLIEVDDERKLRTFYEKRMATEVAADALGEEWKGYVVRISGGNDKQGFPMKQGVLTHGRVRLLLSKGHSCYRPRRTGERKRKSVRGCIVDANLSVLNLVIVKKGEKDIPGLTDTTVPRRLGPKRASRIRKLFNLSKEDDVRQYVVRKPLNKEGKKPRTKAPKIQRLVTPRVLQHKRRRIALKKQRTKKNKEEAAEYAKLLAKRMKEAKEKRQEQIAKRRRLSSLRASTSKSESSQK DIP protein P62754 original sequence version: 64 A61E435884E636AE cyfp1_mouse Cytoplasmic FMR1-interacting protein 1 Cyfip1 Shyc Kiaa0068 Sra1 Specifically Rac1-associated protein 1 protein protein mouse Mus musculus Mouse MAAQVTLEDALSNVDLLEELPLPDQQPCIEPPPSSLLYQPNFNTNFEDRNAFVTGIARYIEQATVHSSMNEMLEEGQEYAVMLYTWRSCSRAIPQVKCNEQPNRVEIYEKTVEVLEPEVTKLMNFMYFQRNAIERFCGEVRRLCHAERRKDFVSEAYLITLGKFINMFAVLDELKNMKCSVKNDHSAYKRAAQFLRKMADPQSIQESQNLSMFLANHNKITQSLQQQLEVISGYEELLADIVNLCVDYYENRMYLTPSEKHMLLKVMGFGLYLMDGSVSNIYKLDAKKRINLSKIDKYFKQLQVVPLFGDMQIELARYIKTSAHYEENKSRWTCASSSSSPQYNICEQMIQIREDHMRFISELARYSNSEVVTGSGRQEAQKTDAEYRKLFDLALQGLQLLSQWSAHVMEVYSWKLVHPTDKYSNKDCPDNAEEYERATRYNYTTEEKFALVEVIAMIKGLQVLMGRMESVFNHAIRHTVYAALQDFSQVTLREPLRQAIKKKKNVIQSVLQAIRKTVCDWETGHEPFNDPALRGEKDPKSGFDIKVPRRAVGPSSTQLYMVRTMLESLIADKSGSKKTLRSSLEGPTILDIEKFHRESFFYTHLINFSETLQQCCDLSQLWFREFFLELTMGRRIQFPIEMSMPWILTDHILETKEASMMEYVLYSLDLYNDSAHYALTKFNKQFLYDEIEAEVNLCFDQFVYKLADQIFAYYKVMAGSLLLDKRLRSECKNQGATIHLPPSNRYETLLKQRHVQLLGRSIDLNRLITQRVSAAMYKSLELAIGRFESEDLTSVVELDGLLEINRMTHKLLSRYLTLDSFDAMFREANHNVSAPYGRITLHVFWELNYDFLPNYCYNGSTNRFVRTVLPFSQEFQRDKQPNAQPQYLHGSKALNLAYSSIYGSYRNFVGPPHFQVICRLLGYQGIAVVMEELLKVVKSLLQGTILQYVKTLMEVMPKICRLPRHEYGSPGILEFFHHQLKDIVEYAELKTVCFQNLREVGNAVLFCLLIEQSLSLEEVCDLLHAAPFQNILPRIHVKEGERVDAKMKRLESKYAPLHLVPLIERLGTPQQIAIAREGDLLTKERLCCGLSMFEVILTRIRTFLDDPIWRGPLPSNGVMHVDECVEFHRLWSAMQFVYCIPVGTHEFTVEQCFGDGLHWAGCMIIVLLGQQRRFAVLDFCYHLLKVQKHDGKDEIIKNVPLKKMVERIRKFQILNDEIITILDKYLKSGDGESTPVEHVRCFQPPIHQSLASS 793986FB417CF19B camk2a_mouse_rna Mouse Camk2a mRNA mrna messenger rna mRNA mrna mouse Mus musculus Mouse drd2_mouse_rna Mouse DRD2 mRNA mrna messenger rna mRNA mrna mouse Mus musculus Mouse fmr1_mouse_rna mrna messenger rna mRNA mrna mouse Mus musculus Mouse if6_mouse Eukaryotic translation initiation factor 6 Eif6 Itgb4bp B4 integrin interactor CAB p27(BBP) protein protein mouse Mus musculus Mouse MAVRASFENNCEVGCFAKLTNAYCLVAIGGSENFYSVFEGELSDAIPVVHASIAGCRIIGRMCVGNRHGLLVPNNTTDQELQHIRNSLPDSVQIRRVEERLSALGNVTTCNDYVALVHPDLDRETEEILADVLKVEVFRQTVADQVLVGSYCVFSNQGGLVHPKTSIEDQDELSSLLQVPLVAGTVNRGSEVIAAGMVVNDWCAFCGLDTTSTELSVVESVFKLNEAKPSTIATSMRDSLIDSLT 35BF0F4C17D5E045 if4e_mouse Eukaryotic translation initiation factor 4E Eif4e eIF-4F 25 kDa subunit protein protein mouse Mus musculus Mouse MATVEPETTPTTNPPPAEEEKTESNQEVANPEHYIKHPLQNRWALWFFKNDKSKTWQANLRLISKFDTVEDFWALYNHIQLSSNLMPGCDYSLFKDGIEPMWEDEKNKRGGRWLITLNKQQRRSDLDRFWLETLLCLIGESFDDYSDDVCGAVVNVRAKGDKIAIWTTECENRDAVTHIGRVYKERLGLPPKIVIGYQSHADTATKSGSTTKNRFVV DIP protein P63073 original sequence version: 68 DIP protein P63073 original sequence version: 67 DIP protein P63073 original sequence version: 87 DIP protein P63073 original sequence version: 119 DIP protein P63073 original sequence version: 85 FC61D0FB337BCD8F fmr1_human Synaptic functional regulator FMR1 FMR1 Fragile X mental retardation protein 1 protein protein human Homo sapiens Human MEELVVEVRGSNGAFYKAFVKDVHEDSITVAFENNWQPDRQIPFHDVRFPPPVGYNKDINESDEVEVYSRANEKEPCCWWLAKVRMIKGEFYVIEYAACDATYNEIVTIERLRSVNPNKPATKDTFHKIKLDVPEDLRQMCAKEAAHKDFKKAVGAFSVTYDPENYQLVILSINEVTSKRAHMLIDMHFRSLRTKLSLIMRNEEASKQLESSRQLASRFHEQFIVREDLMGLAIGTHGANIQQARKVPGVTAIDLDEDTCTFHIYGEDQDAVKKARSFLEFAEDVIQVPRNLVGKVIGKNGKLIQEIVDKSGVVRVRIEAENEKNVPQEEEIMPPNSLPSNNSRVGPNAPEEKKHLDIKENSTHFSQPNSTKVQRVLVASSVVAGESQKPELKAWQGMVPFVFVGTKDSIANATVLLDYHLNYLKEVDQLRLERLQIDEQLRQIGASSRPPPNRTDKEKSYVTDDGQGMGRGSRPYRNRGHGRRGPGYTSGTNSEASNASETESDHRDELSDWSLAPTEEERESFLRRGDGRRRGGGGRGQGGRGRGGGFKGNDDHSRTDNRPRNPREAKGRTTDGSLQIRVDCNNERSVHTKTLQNTSSEGSRLRTGKDRNQKKEKPDSVDGQQPLVNGVP DIP protein Q06787 original sequence version: 68 F853D6C82E3489B9 7-methyl-gtp 7-methyl-GTP small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) map1b_mouse_mrna Murine Map1b mRNA mrna messenger rna mRNA mrna mouse Mus musculus Mouse fmr1_mouse Synaptic functional regulator FMR1 Fmr1 Fmr-1 Fragile X mental retardation protein 1 homolog protein protein mouse Mus musculus Mouse MEELVVEVRGSNGAFYKAFVKDVHEDSITVAFENNWQPERQIPFHDVRFPPPVGYNKDINESDEVEVYSRANEKEPCCWWLAKVRMIKGEFYVIEYAACDATYNEIVTIERLRSVNPNKPATKDTFHKIKLEVPEDLRQMCAKESAHKDFKKAVGAFSVTYDPENYQLVILSINEVTSKRAHMLIDMHFRSLRTKLSLILRNEEASKQLESSRQLASRFHEQFIVREDLMGLAIGTHGANIQQARKVPGVTAIDLDEDTCTFHIYGEDQDAVKKARSFLEFAEDVIQVPRNLVGKVIGKNGKLIQEIVDKSGVVRVRIEAENEKSVPQEEEIMPPSSLPSNNSRVGPNSSEEKKHLDTKENTHFSQPNSTKVQRVLVVSSIVAGGPQKPEPKAWQGMVPFVFVGTKDSIANATVLLDYHLNYLKEVDQLRLERLQIDEQLRQIGASSRPPPNRTDKEKGYVTDDGQGMGRGSRPYRNRGHGRRGPGYTSGTNSEASNASETESDHRDELSDWSLAPTEEERESFLRRGDGRRRRGGGRGQGGRGRGGGFKGNDDHSRTDNRPRNPREAKGRTADGSLQSASSEGSRLRTGKDRNQKKEKPDSVDGLQPLVNGVP 093DD90D589ED066 if4e_human Eukaryotic translation initiation factor 4E mRNA cap-binding protein eIF-4F 25 kDa subunit EIF4E EIF4EL1 EIF4F protein protein human Homo sapiens Human MATVEPETTPTPNPPTTEEEKTESNQEVANPEHYIKHPLQNRWALWFFKNDKSKTWQANLRLISKFDTVEDFWALYNHIQLSSNLMPGCDYSLFKDGIEPMWEDEKNKRGGRWLITLNKQQRRSDLDRFWLETLLCLIGESFDDYSDDVCGAVVNVRAKGDKIAIWTTECENREAVTHIGRVYKERLGLPPKIVIGYQSHADTATKSGSTTKNRFVV DIP protein P06730 original sequence version: 164 DIP protein P06730 original sequence version: 127 DIP protein P06730 original sequence version: 126 DIP protein P06730 original sequence version: 135 DIP protein P06730 original sequence version: 151 DIP protein P06730 original sequence version: 155 DIP protein P06730 original sequence version: 160 DIP protein P06730-1 original sequence version: 187 DIP protein P06730 original sequence version: 146 B869B8DE615E699D fmr1-eif6-1 4 15 unspecified role unspecified role neutral component neutral component 20 unspecified role unspecified role neutral component neutral component 14 unspecified role unspecified role neutral component neutral component 19 unspecified role unspecified role neutral component neutral component 24 unspecified role unspecified role neutral component neutral component physical association physical association These sedimentation experiments suggest that FMRP, CYFIP, and eIF4E might reside in a common complex, perhaps with mRNA. Fig. 1A, Supp. Fig 1 BABP was also bound but the antibodies used were not specific enough to identify proteins as a specific gene product. fmr1-cyfip1-1 5 22 unspecified role unspecified role bait bait 20 unspecified role unspecified role prey prey mouse-brain Mus musculus brain brain Brain [cerebrum] 15 unspecified role unspecified role prey prey mouse-brain Mus musculus brain brain Brain [cerebrum] 24 unspecified role unspecified role prey prey mouse-brain Mus musculus brain brain Brain [cerebrum] association association When the beads were incubated with total brain cytoplasmic extracts, both FMRP and CYFIP1 were recovered from them but only after specific elution with m7GTP (Figure 1B, lane 3) Fig. 1B, 1F, 4B, Supp. Fig. 3 BABP was also bound but the antibodies used were not specific enough to identify proteins as a specific gene product. Human 4E-BP The FMRP-CYFIP1 complex binds eIF4E in competition with 4E-BP. Interaction was not observed in cells in which the dendritic, non-translatable RNA BC1, has been knocked-out Using Spg1-[3H]GDP and Spg1-[g-35S]GTP, the GDP and GTP dissociation rates for Spg1 were measured as 0.7 min-1 and 0.07 min-1, respectively. Using Spg1-[g-32P]GTP, an apparent first-order rate constant of 0.3 min-1 was found. fmr1-cyfip1-2 8 24 unspecified role unspecified role bait bait 20 unspecified role unspecified role prey prey 15 unspecified role unspecified role prey prey association association A specific interaction among FMRP, eIF4E, and CYFIP1 was detected in wild-type (Figure 1C, lane 4) but not in FMR1 knockout (KO) mice (Figure 1C, lane 3) Fig. 1C BABP was also bound but the antibodies used were not specific enough to identify proteins as a specific gene product. Interaction not observed in Fmr1 -/- mice and not disrupted by RNase A treatment. eif4e-cyfip1-3 5 20 unspecified role unspecified role prey prey mouse-brain Mus musculus brain brain Brain [cerebrum] 15 unspecified role unspecified role prey prey mouse-brain Mus musculus brain brain Brain [cerebrum] 22 unspecified role unspecified role bait bait association association Fig. 1E BABP was also bound but the antibodies used were not specific enough to identify proteins as a specific gene product. eif4e-cyfip1-4 1 20 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 P63073:p.Trp73Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain W A Wed May 02 15:31:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp73ala' Tue Jun 30 16:52:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp73Ala' ecoli Escherichia coli (strain K12) 11 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 direct interaction direct interaction Fig. 3A, 3B, 4A CYFIP1 was precipitated specifically by GST-eIF4E (Figure 3A, compare lane 4 with lanes 2 and 3), 7me_gtp-cyfip1-1 1 22 unspecified role unspecified role bait bait 11 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 Q7L576:p.[Asp724Ala;Arg726Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain D A Fri Jul 22 10:57:56 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg726ala,asp724ala' Wed May 02 15:40:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp724ala,arg726ala' Tue Jun 30 18:05:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asp724Ala;Arg726Ala]' region required to bind necessary binding region required to bind mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Q7L576:p.Lys725Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K E Wed May 02 15:34:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys725glu' Tue Jun 30 18:02:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys725Glu' in vitro In vitro direct interaction direct interaction Fig. 3D From the text, it is probably that the 7me-GTP resin has been pre-treated with mouse brain extract and the CYFIP protein is binding via an interaction with eIF4E - the mutants would there fore effect eIF4E binding - however this is not made clear in the methodology. Residue numbering differs between that given in the paper and UniProtKB - UniProtKB numbering has been used throughout. eif4e-cyfip1-2 5 25 unspecified role unspecified role prey prey 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) 11 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q7L576:p.[Asp724Ala;Arg726Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain D A Fri Jul 22 11:04:01 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg726ala,asp724ala' Wed May 02 15:00:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp724ala,arg726ala' Tue Jun 30 16:43:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asp724Ala;Arg726Ala]' Q7L576:p.Lys725Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K E Wed May 02 16:14:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys725glu' Tue Jun 30 20:20:41 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys725Glu' region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Fig. 3E Human GST-eIF4E was applied to streptavidin beads containing wild-type or mutant CYFIP1 biotinylated peptides Residue numbering differs between that given in the paper and UniProtKB - UniProtKB numbering has been used throughout. eif4e-cyfip1-5 1 21 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 20 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) 11 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 association association Fig. 4A Interaction with FMR1 requires the addition of target mRNA Arc mRNA could not be added as an interactor as the species of origin is not given cyfip1-map1b_rna-1 6 15 unspecified role unspecified role bait bait 23 unspecified role unspecified role prey prey physical association physical association The interaction was decreased in the absence of BC1 RNA or in Fmr -/- knockouts. . Figure 4D (lanes 2 and 3) shows that Map1b mRNA (Bagni and Greenough, 2005) was detected in the CYFIP1 immunoprecipitate. Fig. 4D, 4E eif4e-cyfip1-6 2 20 unspecified role unspecified role neutral component neutral component 15 unspecified role unspecified role neutral component neutral component colocalization colocalization CYFIP1-eIF4E colocalization in dendrites Brain-derived neurotrophic factor (BDNF) Fig. 5A, Supp. Fig. 10 fmr1-cyfip1-3 3 20 unspecified role unspecified role prey prey 24 unspecified role unspecified role prey prey 15 unspecified role unspecified role bait bait association association CYFIP1 was immunoprecipitated under control conditions (lane 2, bottom panel) or after BDNF treatment (lane 6), and its presence with eIF4E and FMRP was analyzed by western blotting. Fig. 5B The yield of coprecipitating eIF4E, normalized for the amount of precipitated CYFIP1, was reduced (by 40%) after 30 min of BDNF treatment, although FMRP levels did not change fmr1-cyfip1-4 5 24 unspecified role unspecified role prey prey mouse-neuron Mus musculus neuronal cell neuron Neuron [Neuronal cell; Neural cell 15 unspecified role unspecified role prey prey mouse-neuron Mus musculus neuronal cell neuron Neuron [Neuronal cell; Neural cell 22 unspecified role unspecified role bait bait 20 unspecified role unspecified role prey prey mouse-neuron Mus musculus neuronal cell neuron Neuron [Neuronal cell; Neural cell association association m7GTP-Sepharose beads were incubated with synaptic extracts; the recovered proteins were immunoblotted for CYFIP1, FMRP, and eIF4E Fig. 6A eif4e-cyfip1-7 3 20 unspecified role unspecified role prey prey 15 unspecified role unspecified role bait bait physical association physical association We then stimulated cortical synaptoneurosomes with BDNF, immunoprecipitated CYFIP1, and examined coprecipitated eIF4E Figs. 6B, 6D, Supp. Fig. 13 Brain-derived neurotrophic factor (BDNF) Group I mGluR agonist DHPG cyfip1-rna-1 9 17 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey 13 unspecified role unspecified role prey prey 18 unspecified role unspecified role prey prey 16 unspecified role unspecified role prey prey 23 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 15 unspecified role unspecified role bait bait association association indeed, around 80% of Map1b and BC1 RNAs, as measured by RT-Q-PCR, was released from the CYFIP1 complex after stimulation (Figure 6C). Fig. 6C, Supp. Fig. 9 Brain-derived neurotrophic factor (BDNF) fmr1-cyfip1-5 7 20 unspecified role unspecified role unspecified role unspecified role 24 unspecified role unspecified role unspecified role unspecified role 15 unspecified role unspecified role unspecified role unspecified role association association To test whether the FMRP-CYFIP1 complex simultaneously interacts with PABP, we used poly(A)-Sepharose beads to isolate PABP and associated factors from brain extracts. Supp. Fig. 4 Protein-A Sepharose also isolated PABP but antibodies were not specific enough to identify a particular gene product. Component binding does not require the presence of Cyfip1 target mRNAs or nonspecific binding of CYFIP1 or FMRP to polyribonucleotides fmr1-cyfip1-6 2 15 unspecified role unspecified role neutral component neutral component 24 unspecified role unspecified role neutral component neutral component colocalization colocalization Supp. Fig. 10 fmr1-eif4e-1 2 20 unspecified role unspecified role neutral component neutral component 24 unspecified role unspecified role neutral component neutral component colocalization colocalization Supp. Fig. 10 fmr1-cyfip1-7 5 11 unspecified role unspecified role prey prey human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 21 unspecified role unspecified role prey prey human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 25 unspecified role unspecified role prey prey human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 22 unspecified role unspecified role bait bait association association CYFIP1 and FMRP still bound eIF4E (Figure 1G, lane 2), Fig. 1G Binding of CYFIP1 to eIF4E does not require functional eIF4G.