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/ Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. J. Biol. Chem. (0021-9258) 2001 Yahalom A., Kim TH., Winter E., Karniol B., von Arnim AG., Chamovitz DA. The authors note that "it must be pointed out that all immunoprecipitates were washed with RIPA buffer, conditions that would not allow identification of weak interactions". chamd@post.tau.ac.il 16-April-2008: Contacted by JYOTI. arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Yahalom A., Kim TH., Winter E., Karniol B., von Arnim AG., Chamovitz DA. The authors note that "it must be pointed out that all immunoprecipitates were washed with RIPA buffer, conditions that would not allow identification of weak interactions". chamd@post.tau.ac.il Accepted 2008-APR-3 by JYOTI. J. Biol. Chem. (0021-9258) 2001 Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. J. Biol. Chem. (0021-9258) 2001 Yahalom A., Kim TH., Winter E., Karniol B., von Arnim AG., Chamovitz DA. The authors note that "it must be pointed out that all immunoprecipitates were washed with RIPA buffer, conditions that would not allow identification of weak interactions". chamd@post.tau.ac.il 16-April-2008: Contacted by JYOTI. arath-root Arabidopsis thaliana root tissue root molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Yahalom A., Kim TH., Winter E., Karniol B., von Arnim AG., Chamovitz DA. The authors note that "it must be pointed out that all immunoprecipitates were washed with RIPA buffer, conditions that would not allow identification of weak interactions". chamd@post.tau.ac.il Accepted 2008-APR-3 by JYOTI. 16-April-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2001 Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. J. Biol. Chem. (0021-9258) 2001 Yahalom A., Kim TH., Winter E., Karniol B., von Arnim AG., Chamovitz DA. The authors note that "it must be pointed out that all immunoprecipitates were washed with RIPA buffer, conditions that would not allow identification of weak interactions". chamd@post.tau.ac.il 16-April-2008: Contacted by JYOTI. 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 Yahalom A., Kim TH., Winter E., Karniol B., von Arnim AG., Chamovitz DA. The authors note that "it must be pointed out that all immunoprecipitates were washed with RIPA buffer, conditions that would not allow identification of weak interactions". chamd@post.tau.ac.il Accepted 2008-APR-3 by JYOTI. 16-April-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2001 Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. J. Biol. Chem. (0021-9258) 2001 Yahalom A., Kim TH., Winter E., Karniol B., von Arnim AG., Chamovitz DA. The authors note that "it must be pointed out that all immunoprecipitates were washed with RIPA buffer, conditions that would not allow identification of weak interactions". chamd@post.tau.ac.il 16-April-2008: Contacted by JYOTI. braob Brassica oleracea var. botrytis (Cauliflower) molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Yahalom A., Kim TH., Winter E., Karniol B., von Arnim AG., Chamovitz DA. The authors note that "it must be pointed out that all immunoprecipitates were washed with RIPA buffer, conditions that would not allow identification of weak interactions". chamd@post.tau.ac.il Accepted 2008-APR-3 by JYOTI. 16-April-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2001 Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. J. Biol. Chem. (0021-9258) 2001 Yahalom A., Kim TH., Winter E., Karniol B., von Arnim AG., Chamovitz DA. The authors note that "it must be pointed out that all immunoprecipitates were washed with RIPA buffer, conditions that would not allow identification of weak interactions". chamd@post.tau.ac.il 16-April-2008: Contacted by JYOTI. arath-root Arabidopsis thaliana root tissue root fluorescence imaging fluorescence microscopy fluorescence imaging antibody detection identification by antibody antibody detection Yahalom A., Kim TH., Winter E., Karniol B., von Arnim AG., Chamovitz DA. The authors note that "it must be pointed out that all immunoprecipitates were washed with RIPA buffer, conditions that would not allow identification of weak interactions". chamd@post.tau.ac.il Accepted 2008-APR-3 by JYOTI. 16-April-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2001 Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. J. Biol. Chem. (0021-9258) 2001 Yahalom A., Kim TH., Winter E., Karniol B., von Arnim AG., Chamovitz DA. The authors note that "it must be pointed out that all immunoprecipitates were washed with RIPA buffer, conditions that would not allow identification of weak interactions". chamd@post.tau.ac.il 16-April-2008: Contacted by JYOTI. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Yahalom A., Kim TH., Winter E., Karniol B., von Arnim AG., Chamovitz DA. The authors note that "it must be pointed out that all immunoprecipitates were washed with RIPA buffer, conditions that would not allow identification of weak interactions". chamd@post.tau.ac.il Accepted 2008-APR-3 by JYOTI. 16-April-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2001 Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. J. Biol. Chem. (0021-9258) 2001 Yahalom A., Kim TH., Winter E., Karniol B., von Arnim AG., Chamovitz DA. The authors note that "it must be pointed out that all immunoprecipitates were washed with RIPA buffer, conditions that would not allow identification of weak interactions". chamd@post.tau.ac.il 16-April-2008: Contacted by JYOTI. arath-leaf Arabidopsis thaliana leaf leaf leaf fluorescence imaging fluorescence microscopy fluorescence imaging antibody detection identification by antibody antibody detection Yahalom A., Kim TH., Winter E., Karniol B., von Arnim AG., Chamovitz DA. The authors note that "it must be pointed out that all immunoprecipitates were washed with RIPA buffer, conditions that would not allow identification of weak interactions". chamd@post.tau.ac.il Accepted 2008-APR-3 by JYOTI. 16-April-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2001 eif3b-braob-pro Eif3b protein Cauliflower protein protein braob Brassica oleracea var. botrytis (Cauliflower) An amino acid sequence which is not represented in UniProt This protein is a homologue of Eif3b in arabidopsis csn1_arath COP9 signalosome complex subunit 1 Constitutive photomorphogenesis protein 11 Protein FUSCA 6 CSN1 COP11 FUS6 At3g61140 T20K12.40 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MERDEEASGPMMEMCTNGGEETSNRRPIISGEPLDIEAYAALYKGRTKIMRLLFIANHCGGNHALQFDALRMAYDEIKKGENTQLFREVVNKIGNRLGEKYGMDLAWCEAVDRRAEQKKVKLENELSSYRTNLIKESIRMGYNDFGDFYYACGMLGDAFKNYIRTRDYCTTTKHIIHMCMNAILVSIEMGQFTHVTSYVNKAEQNPETLEPMVNAKLRCASGLAHLELKKYKLAARKFLDVNPELGNSYNEVIAPQDIATYGGLCALASFDRSELKQKVIDNINFRNFLELVPDVRELINDFYSSRYASCLEYLASLKSNLLLDIHLHDHVDTLYDQIRKKALIQYTLPFVSVDLSRMADAFKTSVSGLEKELEALITDNQIQARIDSHNKILYARHADQRNATFQKVLQMGNEFDRDVRAMLLRANLLKHEYHARSARKL C2A980E744B23A90 csn7_arath COP9 signalosome complex subunit 7 Protein FUSCA 5 FUS5 At1g02090 T7I23.24 T7I23.25 CSN7 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDIEQKQAEIIDQLVKRASTCKSEALGPLIIEATSHPSLFAFSEILALPNVAQLEGTTDSVYLDLLRLFAHGTWGDYKCNATRLPHLSPDQILKLKQLTVLTLAESNKVLPYDTLMVELDVSNVRELEDFLINECMYAGIVRGKLDQLKRCFEVPFAAGRDLRPGQLGNMLHTLSNWLNTSENLLISIQDKIKWADNMSEMDKKHRKEAEEGVEEVKKSLSMKGDVDIRGNKEMFGEPSGVMDYEEDGIRPKRRRHPVTR CBBDA06097E44689 csn8_arath COP9 signalosome complex subunit 8 Constitutive photomorphogenesis protein 9 Protein FUSCA 7 CSN8 COP9 FUS7 At4g14110 dl3095c protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDLSPVKEALAAKSFDKIADICDTLMLQVASEGIEYHDDWPYAIHLLGYFYVDDCDSARFLWKRIPTAIKERKPEVVAAWGIGQKLWTHDYAGVYEAIRGYDWSQEAKDMVAAFSDLYTKRMFQLLLSAYSTITIHDLALFLGMTEDDATTYVVENGWTVDAASQMASVKKQAVKREQKVDSSKLQRLTEYVFHLEH D182ABACE4238759 eif3c_arath Eukaryotic translation initiation factor 3 subunit C TIF3C1 F18O21_110 At3g56150 eIF3 p110 p105 Eukaryotic translation initiation factor 3 subunit 8 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MTSRFFTQVGSESEDESDYEVEVNEVQNDDVNNRYLQSGSEDDDDTDTKRVVKPAKDKRFEEMTYTVDQMKNAMKINDWVSLQENFDKVNKQLEKVMRITEAVKPPTLYIKTLVMLEDFLNEALANKEAKKKMSTSNSKALNSMKQKLKKNNKLYEDDINKYREAPEVEEEKQPEDDDDDDDDDDEVEDDDDSSIDGPTVDPGSDVDEPTDNLTWEKMLSKKDKLLEKLMNKDPKEITWDWVNKKFKEIVAARGKKGTARFELVDQLTHLTKIAKTPAQKLEILFSVISAQFDVNPGLSGHMPINVWKKCVLNMLTILDILVKYSNIVVDDTVEPDENETSKPTDYDGKIRVWGNLVAFLERVDTEFFKSLQCIDPHTREYVERLRDEPMFLALAQNIQDYFERMGDFKAAAKVALRRVEAIYYKPQEVYDAMRKLAELVEEEEETEEAKEESGPPTSFIVVPEVVPRKPTFPESSRAMMDILVSLIYRNGDERTKARAMLCDINHHALMDNFVTARDLLLMSHLQDNIQHMDISTQILFNRTMAQLGLCAFRAGMITESHSCLSELYSGQRVRELLAQGVSQSRYHEKTPEQERMERRRQMPYHMHLNLELLEAVHLICAMLLEVPNMAANSHDAKRRVISKNFRRLLEISERQAFTAPPENVRDHVMAATRALTKGDFQKAFEVLNSLEVWRLLKNRDSILDMVKDRIKEEALRTYLFTYSSSYESLSLDQLAKMFDVSEPQVHSIVSKMMINEELHASWDQPTRCIVFHEVQHSRLQSLAFQLTEKLSILAESNERAMESRTGGGGLDLSSRRRDNNQDYAGAASGGGGYWQDKANYGQGRQGNRSGYGGGRSSGQNGQWSGQNRGGGYAGRVGSGNRGMQMDGSSRMVSLNRGVRT 84EA72DDE49A05C5 eif3e-braob-pro Eif3e protein Cauliflower protein protein braob Brassica oleracea var. botrytis (Cauliflower) An amino acid sequence which is not represented in UniProt Homologue of Eif3e protein in arabidopsis csn7-braob-pro CSN7 protein Cauliflower protein protein braob Brassica oleracea var. botrytis (Cauliflower) An amino acid sequence which is not represented in UniProt Homologue sof CSN7 protein in arabidopsis q9m4t6_arath Eukaryotic translation initiation factor 3 subunit B eIF-3-eta eIF3 p110 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAAVVDIDVLTAQLGINWSDVNLDSIQLPPGNDFGIESDDESVYQDDQSEFDTGFGNIIVVDHLPVVPKEKFEKLEGVVKKIYNQLGVIKENGLLMPVDPDTKMTLGYCFIEFNTPQEAQNAKEKSHGYKLDKSHIFAVNMFDDFDRLMNVKEEWEPPQERPYVPGENLQKWLTDDKARDQLVIRFGHDTEVYWNDARQKKPEPVHKRSYWTESYVQWSPIGTYLVTLHKQGAAVWGGADTFTRLMRYQHSMVKLVDFSPGEKYLVTYHSQEPSNPRDASKVEIKVFDVRTGRMMRDFKGSADEFSIGGPGGVAGASWPVFRWACGKDDKYFAKLSKNTISVYETETFSLIDKKSMKVDNVVDICWSPTDSILSLFVPEQGGGNQPAKVALVQIPSKVELRQKNLFSVSDCKMYWQSSGEYLAVKVDRYTKTKKSTYSGFELFRIKERDIPIEVLELDNKNDKIIAFAWEPKGQRFAGIHGDQPNRDVSFYSIKTGTKTLERVSKLATLKAKQAKCPLLVAPQASTSFLAGLKGFNGQLEFFNVDELETMATAEHFMATDIEWDPTGRYVATSVTTVHEMENGFTIWSFNGNMVYRILKDHFFQLAWRPRPASFLTAEKEEEIAKNLRNYSKRYEAEDQDVSLLLSEQDREKRRALNEEWQKWVMQWKSLHEEEKLVRQNLRDGEVSDVEEDEYEAKEVEFEDLIDVTEEIVQELM 516FC664E2D2CFA2 eif3e_arath Eukaryotic translation initiation factor 3 subunit E TIF3E1 Eukaryotic translation initiation factor 3 subunit 6 INT6 F28O9.140 At3g57290 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEESKQNYDLTPLIAPNLDRHLVFPIFEFLQERQLYPDEQILKSKIQLLNQTNMVDYAMDIHKSLYHTEDAPQEMVERRTEVVARLKSLEEAAAPLVSFLLNPNAVQELRADKQYNLQMLKERYQIGPDQIEALYQYAKFQFECGNYSGAADYLYQYRTLCSNLERSLSALWGKLASEILMQNWDIALEELNRLKEIIDSKSFSSPLNQVQNRIWLMHWGLYIFFNHDNGRTQIIDLFNQDKYLNAIQTSAPHLLRYLATAFIVNKRRRPQLKEFIKVIQQEHYSYKDPIIEFLACVFVNYDFDGAQKKMKECEEVIVNDPFLGKRVEDGNFSTVPLRDEFLENARLFVFETYCKIHQRIDMGVLAEKLNLNYEEAERWIVNLIRTSKLDAKIDSESGTVIMEPTQPNVHEQLINHTKGLSGRTYKLVNQLLEHTQAQATR A9E6656E51AA1483 csn8-eif3c 1 11 unspecified role unspecified role bait bait 12 unspecified role unspecified role prey prey physical association physical association Figure 1A eIF3c also coimmunoprecipitated with CSN8 csn1-eif3c 1 12 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association Figure 1A eIF3c also coimmunoprecipitated with CSN8 and CSN1. eif3-eif3c 1 16 unspecified role unspecified role bait bait 12 unspecified role unspecified role prey prey physical association physical association Figure 1A As seen in Fig. 1A, eIF3c coimmunoprecipitated with eIF3e and eIF3b, indicating that these three proteins are components of the same protein complex, eif3b-eif3c 1 12 unspecified role unspecified role prey prey 15 unspecified role unspecified role bait bait physical association physical association Figure 1A As seen in Fig. 1A, eIF3c coimmunoprecipitated with eIF3e and eIF3b, indicating that these three proteins are components of the same protein complex, eif3-csn7 1 10 unspecified role unspecified role prey prey 16 unspecified role unspecified role bait bait physical association physical association Figure 1B eIF3e coimmunoprecipitated a core component of the COP9 signalosome, CSN7, in extracts from both wild type and cop9 mutants, The eIF3e protein coimmunoprecipitated a core component of the COP9 signalosome, CSN7, in extracts from both wild type and cop9 mutants, eif3-csn7-1 3 16 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait physical association physical association Figure 2A The interaction of eIF3e with CSN7 clearly activated b-galactosidase activity indicating a direct interaction between eIF3e and CSN7 and further supporting the immunoprecipitation data. eif3-eif3c-1 3 12 unspecified role unspecified role bait bait 16 unspecified role unspecified role prey prey physical association physical association Figure 2A eIF3e also interacted with eIF3c, suggesting that the immunoprecipitation of eIF3c and eIF3e is a result of direct interaction between these proteins. eif3c-eif3b 3 12 unspecified role unspecified role prey prey 15 unspecified role unspecified role bait bait physical association physical association Figure 2B Despite this, the results clearly show a direct interaction between eIF3b and eIF3c. eif3-eif3c-2 6 12 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 ecoli Escherichia coli (strain K12) 16 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Figure 3B As shown in Fig. 3A the amount of GST-eIF3e protein that was �pulled down� increased as the amount of His6-CSN7 protein increased, whereas the control failed to pull down GST-eIF3e. Similar results were obtained in a pull-down experiment using equal amounts of His6-eIF3c and increasing amounts of GST-eIF3e on glutathione-Sepharose beads (Fig. 3B). csn7-eif3 6 10 unspecified role unspecified role bait bait 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) 16 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) direct interaction direct interaction Figure 3A As shown in Fig. 3A the amount of GST-eIF3e protein that was �pulled down� increased as the amount of His6-CSN7 protein increased, whereas the control failed to pull down GST-eIF3e. Similar results were obtained in a pull-down experiment using equal amounts of His6-eIF3c and increasing amounts of GST-eIF3e on glutathione-Sepharose beads (Fig. 3B). eif3-csn7-2 7 16 unspecified role unspecified role neutral component neutral component 10 unspecified role unspecified role neutral component neutral component colocalization colocalization Figure 5 The localization of eIF3e overlapped that of the CSN7 protein, which was found in both nucleus and cytoplasm of root and leaf protoplasts The localization of eIF3e overlapped that of the CSN7 protein, which was found in both nucleus and cytoplasm of root and leaf protoplasts eif3-csn7-3 5 10 unspecified role unspecified role neutral component neutral component 16 unspecified role unspecified role neutral component neutral component colocalization colocalization Figure 5 The localization of eIF3e overlapped that of the CSN7 protein, which was found in both nucleus and cytoplasm of root and leaf protoplasts The localization of eIF3e overlapped that of the CSN7 protein, which was found in both nucleus and cytoplasm of root and leaf protoplasts eif3-csn7-eif3b 2 16 unspecified role unspecified role neutral component neutral component 15 unspecified role unspecified role neutral component neutral component 10 unspecified role unspecified role neutral component neutral component physical association physical association Figure 6 eIF3e and eIF3b are part of a protein complex larger than the COP9 signalosome. eif3-csn7-eif3b-1 4 8 unspecified role unspecified role neutral component neutral component 14 unspecified role unspecified role neutral component neutral component 13 unspecified role unspecified role neutral component neutral component physical association physical association Figure 6 To determine if eIF3e and eIF3b are found in a large molecular weight complex, total soluble proteins from cauliflower buds and Arabidopsis roots were separated by gel filtration chromatography, and the fractions were subjected to Western blot analysis with alpha -eIF3e and alpha -eIF3b antibodies. In both tissues, the peak elutions of eIF3e and eIF3b were in a large molecular weight species, which is larger than the COP9 signalosome, as shown by the elution profile of CSN7 (Fig. 6).